Microfluidic Drug Discovery
Microfluidic automated fluidic workflows as valuable tools for drug discovery
- Increased throughput and reproducibility
- Minimal reagent consumption
- Fast reaction times
- Automated fluidic workflow
Main applications in microfluidic drug discovery
Target selection and validation (drug synthesis)
When developing a drug target, protein structural and affinity studies are needed for target selection and validation, and protein interactions within cells must be studied. Single-cell protein quantitation, protein analysis in nanoliter droplets, and high-sensitivity ligand-binding interactions can be performed using microfluidic devices.
Hit identification and optimization (drug screening)
In microfluidics for drug development, the potential pool size of drug candidates is estimated to be of the order of 1063. Microfluidic drug screening devices, including high-throughput microfluidic multiplexed systems that contain thousands of micrometer chambers, microwell arrays, or microvalves perform high-throughput screening studies with higher sensitivity and shorter reaction times while decreasing reagents volumes and costs.
Preclinical studies (drug evaluation)
In recent years, alternatives to animal experimentation have become a research hotspot. In microfluidic drug discovery, organs-on- chips and organoids are widely used as alternatives as they emulate the architectural and functional complexity of native organs. These systems also enable the exploration of facets of human disease and development that are not accurately recapitulated by animal models.
Resources
Flow Control Performance and Enhanced Automation for Microfluidic Drug Discovery Applications
- Drug synthesis and droplet microfluidics: When it comes to droplet or particle generation, having control of the fluid delivery system is important. During droplet or particle production, the flow rate of each phase must remain constant and stable to allow the production of monodisperse droplets.
- As experts in fluid control for droplet microfluidics, Fluigent provides droplet generation packages and platforms for researchers. These seamlessly generate micrometer droplets for a wide range of applications. Custom systems are also available to integrate technological and industrial devices.
- Multiplexing for drug screening: More than twenty reagents can be tested simultaneously when performing multiplexing applications on microfluidic devices. In microfluidic drug discovery, automated fluidic workflows are a prerequisite for drug screening. Fluigent developed the Aria to automate multiple fluid deliveries. Reproducibility is improved through complete automation of the fluidic protocol. Fluigent is also able to develop a custom system to fit user specifications and requirements.
- Optimized shear stress and recirculation for organ on a chip studies and cell culture: When performing drug evaluation on 2D/3D cell culture and organs-on-chips, cell culture should be optimized through constant perfusion to enable nutrients/oxygen renewal and by inducing adequate passive stimulation through shear flow as the effect is substantial on cell properties. Sterility on the fluidic path is also a prerequisite. Fluigent develops fluid recirculating systems that perform unidirectional fluid recirculation, ensuring improved and more stable flow rates compared to peristaltic pumps. The system can run for 10 days. Custom systems are also available for microfluidic drug discovery technology integration into industrial devices.
Fluigent Solutions as an Alternative to Syringe Pumps for Better Performance and Automation
Flow rate stability and responsiveness is critical for the microfluidics for drug development applications. Syringe pumps are commonly used during this process. Depending on the model in use, syringe pumps show limited flow control, and the actual flow rate cannot be monitored. In addition, injection volumes are limited by the syringe, and automation can thus be difficult. An alternative to syringe pumps is pressure-based flow controllers. These show high-precision flow control, fast reaction time, and flow monitoring: parameters which are paramount for microfluidic drug discovery applications.
The benefits of choosing Fluigent for your flow control system:
- Best in class stability: < 0.5% thanks to our field-proven, patented FASTAB™ technology allowing optimal flow control with the robustness required in demanding industrial environments
- Workflow automation becomes straightforward with the Fluigent SDK and software included in the system
- Our engineering team members are experts in microfluidic design, mechanical and software integration, and biology applications
Related resources
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Table of contents
I. Droplet-based microfluidics, a microfluidic chip application
Microfluidic generation of droplets has attracted a lot of interest, enabling high throughput experiments by generating millions of micro-reactors and chambers in a few seconds. This microfluidic chip application method produces highly monodispersed droplets of very small volumes (μL to fL) of fluids with high frequency (up to hundreds of kHz), providing better control of processes like mixing, encapsulation, sorting, and sensing. Microfluidic-based droplets have many diverse and varied applications, such as particle synthesis3 and chemical analysis4. Highly controlled droplet production also enables single-cell analysis or drug testing 5,6.
Microfluidic-based droplet generation and control allow for:
- Highly monodispersed (<2% size variation) droplet production, with potentially high generation rate (up to hundreds of kHz)
- Highly reproducible complex structures (water-in-oil-in-water emulsions, multiple encapsulations…)
- Single droplet manipulation as an individual pL scale biochemical reactor
- Miniaturization of production and bioanalytical devices
With these characteristics, droplet microfluidics has a large value, including bio(chemical) analysis, and nano and microscale generation of materials7. Several microfluidic chip designs exist to generate droplets for a various field of applications. A common design is the T-junction, where the dispersed phase is injected from a channel that is perpendicular to the channel carrying the continuous phase (figure 1).
Application of microfluidic chip using droplet microfluidic concepts, components, and processes are now being adopted and leveraged by end-users to enable new science and innovation. Real-world success is now evidenced through a range of mainstream commercial products that are applied to key biological and healthcare-related problems (e.g., 10X Genomics, Drop-seq, and nucleic acid quantification via Droplet DigitalTM PCR systems)8.
Today, it is possible to produce multiple emulsions with complex droplet morphologies. Production of multi-cored droplets (droplets that contain a controlled number of inner droplets at one or more hierarchical levels), Janus droplets (i.e. biphasic or triphasic droplets with two or three physically and chemically distinct surface domains) is now possible using droplet microfluidics9 (figure 2).
Figure 2: Types of multiple emulsions from the simple encapsulation (a), double emulsion (b), and so on (c,d) to bi- and tri-phasic structures (e to h), multiple encapsulations (I to k), and hybrid microjet achievable using droplet-based microfluidics9
Microfluidic chip application for the production of highly reproducible PLGA microparticles
In recent years, biodegradable microspheres/microparticles have gained widespread importance in the delivery of bioactive agents10. The copolymers of Poly (D, L-lactic-co-glycolic acid) (called PLGA)/poly (lactic acid) PLA microparticles are one of the most successful new drug delivery systems (DDS) in labs and clinics. Because of their biocompatibility and biodegradability, they can be used in various areas, such as long-term release systems, vaccine adjuvant, and tissue engineering11.
In PLGA microparticle production for drug release and delivery, microparticle size is a key parameter as it is directly related to the microparticles degradation rate and the drug release rate12. Although PLGA microparticle synthesis appears to be a successful drug delivery system, the current processes and tools to produce PLGA microparticles have many limitations, such as wide microparticle size distribution, poor repeatability, and aggressive chemical preparation conditions11. To solve these problems, droplet-based microfluidics application chip offers an efficient method for improvement.
Fluigent provides the Raydrop: a new breakthrough technology leading to outstanding particle size monodispersity and production flexibility. The Raydrop works as a co-flow focusing principle (figure 3). The nozzle and outlet capillary are aligned in a continuous phase chamber, the dispersed phase comes through the nozzle to create the microparticles into the continuous phase and exits by the outlet insert. Using this application of microfluidic chip method, PLGA microparticles ranging from 15 to 50 µm diameters have been successfully generated.
The PLGA microparticle production station allows excellent reproducibility and significantly improved monodispersity (CV < 2%) as compared to other methods on the market. It allows one to continuously produce PLGA microparticles (up to 10 000 droplets/s) without unwanted interruption for long-term experiments.
Apllication notes:
A microfluidic chip application for single-cell mRNA-seq sequencing using droplets: Drop-seq technology
The production of highly monodispersed emulsions or more complex structures (water-in-oil-in-water emulsions, multiple encapsulations…) makes microfluidic chip applications an excellent approach for single-cell analysis or single-cell culture. The technique allows for droplet-based single-cell RNA-sequencing, such that one can characterize complex tissues with many cell types and states under diverse conditions. One of the pioneering microfluidic chip application methods is Drop-Seq technology, which entraps a single cell and a single primer-barcoded bead in each droplet (figure 4).
Cells are thus separated into nanoliter-sized aqueous droplets, with a different barcode associated with each cell’s mRNAs. The primers on beads contain a barcode consisting of three sequences. One sequence is for PCR amplification and is common to all the beads. The second sequence consists of hundreds of individual primers that also share the same ‘‘cell barcode’’.
Finally, the third part has different unique molecular identifiers (UMI), enabling mRNA transcripts to be digitally counted13 (figure 5). The droplets are sequenced altogether, allowing quick profiling of thousands of individual cells from a heterogeneous population.
The power of this microfluidic chip application technology resides in the fact that during sequencing, one can distinguish where the original information came on a cell to cell basis.
This allows one to make a gene expression map of the cell, or even to distinguish cell populations within a tissue.
Apllication notes:
II. Microfluidic cell culture for a better cell behavior understanding
Microfluidic cell culture is another application of microfluidic chip that has significant advantages over macroscopic culture in flasks, dishes, and well-plates14 (figure 6). The microfluidic chip fabrication process allows great flexibility in the design of microfluidic devices, permitting one to understand and control interactions between cells, substrates, and the surrounding medium, physically as well as biochemically15.
This microfluidic chip application technology offers new possibilities to accurately reproduce the cellular environment and enables the analysis of biological processes that were not accessible before. Morphology-wise, chips can be structured at the cell scale to reproduce the mechanical constraints experienced by cells. Biochemically, stable gradients can be implemented with a high spatial resolution (typically, micrometer resolution).
Finally, constant perfusion enables the continuous renewal of nutrients and oxygen to promote cell growth and maintain optimal activity during long-term cell culture. Cost reduction due to volume reduction is also a major benefit15.
Microfluidic chip application model of a tumor microenvironment
The physical microenvironment of tumors is characterized by heterotypic cell interactions and physiological gradients of nutrients, waste products, and oxygen. This tumor microenvironment has a major impact on the biology of cancer cells and their response to chemotherapeutic agents. Despite this, most in vitro cancer research still relies primarily on cells grown in 2D and in isolation in nutrient and oxygen-rich conditions.
Ayuso et al. presented an easy-to-use microfluidic chip application device that can mimic the three-dimensional architecture of multicellular spheroids, while at the same time generating a visible, live “tumor slice” that allows easy monitoring of cells in different regions of the microenvironment in real-time as well as their response to different drugs17 (figure 7).
In this application of microfluidic chip setup, tumor cell behavior in different regions of the microdevice was studied and analyzed in conjugation with measurements of hypoxia and glucose concentrations across the device. The differential cellular response to several well-known drugs in different parts of the microdevice emphasizes the potential of this technology for analyzing the impact of microenvironmental parameters on drug response.
The figure presents microdevices in a Petri dish containing a central culture chamber and 6 channels. To better understand how the chip works, picture B shows one microdevice filled with (yellowish) collagen hydrogel flowing to the microchamber from the right middle channel and blue-colored water perfused through the two lateral microchannels.
In experimental conditions, the culture medium perfused through the lateral microchannels provides nutrients and oxygen creating physiological gradients across the device. Cells near the ‘surrogate’ blood vessels are viable, whereas oxygen-poor cells in the center of device start to die creating a ‘necrotic core’ similar to the necrotic regions of tumors. It is possible to monitor cells with fluorescent dye in microdevice17 (picture D).
Application notes & expertises:
- Assess Cell Proliferation Using Pressure as a Tool
- Creating a Microfluidic Cancer-on-Chip Platform
- Cancer Cell Analysis Made Easy with Aria: cell Capture and Labeling
- Passive and active mechanical stimulation in microfluidic systems
- Mimicking in-vivo environments: biochemical and biomechanical stimulation
III. Organ on a chip, a cutting edge application of microfluidic chip
Many efforts are devoted to the development of cancer metastasis models that can help in understanding the disease and the development of innovative therapeutic strategies. Current in vitro and in vivo cancer models are incapable of satisfactorily predicting the outcome of various clinical treatments on patients18. Therefore, new application of microfluidic chip methods and approaches for drug discovery and health research are being developed. The concept of mimicking the organ-level function of human physiology or disease using cells inside a microfluidic chip application setup was first published in 2004. In 2010 that the term organ-on-a-chip (OOAC) was invented by Ingber, et. al., who developed a microfluidic chip model to capture organ-level functions of the human lung19.
Microfluidic chip applications enable one the unique ability to control the cellular microenvironment with high spatiotemporal precision and to present cells with mechanical and biochemical signals in a more physiologically relevant context19. The manipulation of the micro-liter volumes of liquids has made these models a platform where scaling, and dynamic crosstalk between cells can be achieved. Microfluidic chips can now use geometries and structures to permit the use of physiological length scales, concentration gradients, and the mechanical forces generated by fluid flow to mimic the in vivo microenvironment experienced by cells20.
These biomimetic applications platforms overcome many drawbacks encountered with conventional tissue culture models. OOAC engineering microfluidic chip application has attracted enormous interest and attention from the pharmaceutical industry, regulatory agencies, and even national defense agencies. This is demonstrated by the increase of OOAC research papers and by the emergence of at least 28 organ-on-a-chip companies in less than seven years21.
A microfluidic chip design to reconstitute organ-level lung functions
To demonstrate that it is possible to engineer a microsystem that replicates the complex physiological functionality of living organs, Huh et al. developed a multifunctional microdevice that reproduces key structural, functional, and mechanical properties of the human alveolar-capillary interface, which is the fundamental functional unit of the living lung19. The microfluidic chip application device consisted of compartmentalized PDMS microchannels to form an alveolar-capillary barrier on a thin, porous, flexible PDMS membrane coated with the extracellular matrix and human alveolar epithelial cells and human pulmonary microvascular endothelial cells is cultured on opposite sides of the membrane (figure 8).
In fact, the device recreates physiological breathing movements (shown in Figure 8.B) by applying vacuum to the side chambers and causing mechanical stretching of the PDMS membrane forming the alveolar-capillary barrier. The device is made of 3 PDMS layers that are bonded to form two sets of three parallel microchannels. (Figure 8.C) PDMS etchant is flowed through the side channels in order to form the two large side chambers. (Figure 8.D). Figure 8.E represents an image of an actual lung-on-a-chip microfluidic device19.
To put it in a nutshell, air is subsequently introduced into the compartment to create an air-liquid interface to mimic the lining of the alveolar air space19 .
Using this microfluidic chip application platform, the authors demonstrated that breathing motions, simulated by the organ-on-chip platform, might greatly accentuate the proinflammatory activities of silica nanoparticles and contribute substantially to the development of acute lung inflammation19. This behavior could not be determined using existing in vitro models.
Application notes & expertises:
- Peristaltic Pump vs Pressure-Based Microfluidic Flow Control for Organ on Chip applications
- Long-term fluid recirculation system for Organ-on-a-Chip applications
- CNRS/UTC: study of a liver-on-a-chip model
- Development of a human gut-on-chip to assess the effect of shear stress on intestinal functions
- Cartilage-on-a-chip, an example of complex mechanical stimulation using Fluigent’s technology
- Creating a Microfluidic Cancer-on-Chip Platform using Fluigent’s High Throughput Cell Perfusion Pack
IV. Particle and cell sorting applications using cell sorter chips
Efficiently isolating and organizing cells from complex mixtures is a crucial task in various fields such as biology, biotechnology, and medicine. Microfluidic chips play a pivotal role in this process, commonly employed to enrich or purify cell samples, thereby enhancing efficiency in research and development22.
Traditionally, optical methods like FACS (Fluorescent Activated Cell Sorting) are used for cell detection. FACS utilizes a laser beam, with the scattered light providing characteristic information about cells and their components. Automated and robust, FACS platform has been a gold standard for cell sorting. However, current commercial platforms face limitations in sample throughput and processing speeds, posing challenges for generating clinical-scale samples.
In contrast, platforms with microfluidic chip offer affordability, simplicity, and a smaller footprint. These chips employ various techniques for cell sorting, each with specific speeds and efficiencies. The chip’s dimensions accommodate diverse cell sorters, ranging from large-volume to precise single-cell sorters.
Additionally, microfluidic cell sorting can integrate various fluidic operations within a single chip, making it versatile for lab-on-a-chip applications, diagnostics, and therapeutics. This approach holds promise in both academic and industrial labs.
Cell sorter in microfluidic devices relies on determining specific cell parameters, such as size, shape, density, or surface markers. Heterogeneous cell solutions are injected into the microfluidic chip, where cells with different properties experience varying forces, leading to their separation into different streamlines and exits.
Microfluidic cell sorting can be categorized into three categories:
- Fluorescent label-based,
- Bead-based
- Label-free cell sorting22
Label-free sorting is perhaps the most studied and comprises both active systems (relying on external fields for sorting) and passive systems that don’t use fluorescent labels or beads. Instead, these methods leverage inherent differences in cellular morphology between cell groups 22.
Inertial microfluidics for continuous particle separation in spiral microchannels
Kuntaegowdanahalli et al. developed a simple inertial microfluidic device for continuous multi-particle separation, using the Dean-coupled inertial migration principle in spiral microchannels.
In this innovative design, dominant inertial forces, combined with the Dean rotational force arising from the microchannel’s curved geometry, cause particles to settle at a single equilibrium position near the inner microchannel wall. The specific position of particle equilibrium is determined by the ratio of inertial lift to Dean drag forces.
The researchers applied this principle to create a spiral lab-on-a-chip, showcasing size-dependent particle focusing at distinct equilibrium positions across the microchannel cross-section from a multi-particle mixture23. Randomly dispersed particles equilibrate at different positions along the inner wall of the spiral microchannel, influenced by lift and drag forces (see Figure 9 – insert 2). As the particles reach the end of the spiral, they align and separate into different channels (see Figure 9 – insert 3).
A notable advantage of this microfluidic chip application is its high throughput, reaching 1.5 mL/min, achieved without the need for sheath flow or sequential cell manipulation. This feature is particularly beneficial for processing native biological fluids and applications in flow cytometry.
Application notes & expertises:
V. Micromixers an application using microfluidic chips
Micromixers play a crucial role in lab-on-chip devices for various microfluidic chip applications, including drug delivery, sequencing, amplification, and biochemical reactions. They can be broadly classified into two categories based on the actuation method: passive and active.
Passive mixing relies on the microfluidic chip’s geometry and fluid properties without external sources. In laminar flow, typical in microfluidics, mixing primarily occurs through diffusion. This property allows for precise tuning of mixing by employing lamination, where two or more liquids flow in parallel, enabling diffusion to take place. For emproved and faster mixing, chaotic advection can be induced by modifying the microfluidic chip’s geometry, altering channel shapes for splitting, folding, stretching, or disrupting fluid flow.
On the other hand, active mixing involves external perturbation. Various methods are employed in the microfluidic chip application field for active mixing. Dielectrophoresis mixing uses an electric field to move particles toward or away from an electrode, creating chaotic advection. Acoustic wave energy can also mix fluids by generating strong acoustic waves that interfere with each other, creating advection 24. Additionally, adjusting the microfluidic chamber temperature can enhance mixing, as the diffusion coefficient of a liquid is temperature-dependent25.
Submillisecond organic synthesis using a serpentine-shaped microfluidic chip application
In chemical synthesis, it is important to explore the synthetic pathways of an intermediate. To fully observe these pathways, control over its lifetime and mixing time is required.
The reaction mixture had to be well-mixed within the lifetime of the reactive intermediate. An efficient means of prolonging the lifetime is to lower the reaction temperature (-78°C to -100 °C), above the melting point of many organic solvents. Using microfluidic devices, mixing can be extremely fast, with mixing times unattainable by batches26. However, mixing time is increased at low temperatures, as the solvent viscosity exponentially increases.
To circumvent this issue, the authors used a three-dimensional serpentine-shaped microfluidic chip, allowing improved mixing by chaotic advection. The conceptual scheme of the 3D serpentine microchannel fabricated by lamination is represented in figure 10.A.
The figure 10.B represents a detailed scheme for a nanoliter reaction space of rectangular 3D serpentine channels with three inlets and the optical image showing from the top the nanoliter reaction space schematized in B. The optical images of respectively the chip reactor module and assembly are illustrated in Figure 10.D and E.
The utilization of this platform for the application enabled submillisecond mixing.
VI. Microvalves and microfluidic chip applications with reduced internal volume
In the past 10 years, efforts have been devoted to the development of microfluidic platforms capable of performing several assays using programmable fluidic operations within an array of microvalves. Similar to programmable logic circuits where multiple electronic computing routines are executed on a single microdevice, programmable microfluidic platforms have been implemented27, allowing one to perform fluidic operations such as mixing, sampling, washing, and reacting automatically within a single microfluidic chip by modifying the sequence/order of fluidic operations using the software.
The primary advantage of microvalves over their macroscale counterparts is the significantly reduced dead volume, which is important in many microfluidic chip applications that require precise flow control at small flow rates28. They are useful in biological and chemical applications, such as quantitative metabolic biomarker and genetic analysis29,30, protein-based biomarker detection31, or small molecule chemical and environmental analysis32. These microfluidic chip and valve application platforms usually consist of a 2D array of microvalves that permit flow regulation, on/off switching and sealing of liquids, gases, or vacuums33.
Several microvalves have been developed using pneumatic, electrokinetic and electrochemical actuators. Among these mechanisms, pneumatic actuation is often recognized as the most reliable method due to the simplicity of fabrication, ease of use, scalability, reliability, and a high degree of accuracy. Pneumatically actuated microvalves utilize the deflection of an elastomer (typically PDMS) membrane to control fluid flow34.
A fully integrated multilayer microfluidic chemical analyze for automated sample processing, labelling, and analysis
Capillary zone electrophoresis (CZE), is a powerful tool for chemical analysis and is widely used for environmental monitoring, astrobiology, and biosensing32. CZE assays usually require complex and manual sample processing.
Commercial platforms for automated CZE have been implemented to address this concern, but are expensive, and large, thus hardly field deployable in challenging environments. Microfluidic application chips and devices allow miniaturization, automation, and reduction in sample volume requirements for chemical and biochemical sensing.
Using membrane microvalve technology, it is possible to automate metering, transporting, routing, and mixing operations. Kim et al. introduced a microfluidic platform consisting of pneumatically actuated “lifting gate” microvalves integrated with a glass CZE microchip, providing extremely low dead volumes between components.
All the procedures, including buffer filling, labeling, and dilution, can be automated. The microchip was used to analyze diverse compound classes, such as amino acids, and oxidized biomarker compounds, like aldehydes/ketones and carboxylic acids in less than 30 min.
VII. Wearable microfluidics: an emerging application
An emerging application of microfluidic chip, is the use of microfluidic concepts for wearable device applications36. Here, microfluidics presents several key value propositions. The microstructures store or handle fluids and are the core of the sensing device. Using microchannels of the microfluidic chip, precise liquid amounts can be manipulated, allowing for highly accurate and reliable devices and being useful for bodily fluids that are often secreted or extracted in limited quantities.
Also, wearable microfluidic devices could also stock a specific drug for precise dispensing at controlled intervals. Innovations in flexible microfluidics and electronics have led to numerous applications. Typically, a wearable microfluidic device will collect a fluid, transfer it to a localized site where detection or measurement is performed. Blood and sweat are common analytes as they provide insights into physiological states such as temperature, pH, and hydration36. Wearable microfluidics finds applications in the pharmaceutical, food, sportswear, and cosmetic fields.
A wearable microfluidic device for the capture, storage and colorimetric sensing of sweat
A wearable microfluidic device for the capture, storage and colorimetric sensing of sweat
As mentioned previously, sweat is an analyte of interest because of its rich content of important biomarkers. It is easy to collect compared to blood. In situ quantitative analysis of sweat is of great interest for monitoring physiologic health status (for example, hydration state) and for the diagnosis of disease37.
Existing systems for sweat collection and analysis are confined to laboratories, where standard analytical technologies can be performed. Though highly precise, the analysis is time-consuming and costly.
To address this issue, Kho et al. developed a soft wearable microfluidic system than can directly harvest sweat from pores on the surface of the skin37.
The device routes the sample to different channels and reservoirs for multiparametric sensing of markers of interest, with options for wireless interfaces to external devices for image capture and analysis.
The device can measure total sweat loss, pH, lactate, chloride, and glucose concentrations by colorimetric detection using wireless data transmission. As it is a simple, low-cost, and fast analysis point of care device, it could be used to accumulate data from individual users over time, and this could serve as an analytical approach for interpreting trends in marker concentrations, potentially providing warning signs when performing physical activity.
Conclusion
Since the introduction of microfluidics, the scope of microfluidic chip applications has kept extending over the years. The first applications were focused on analytical chemistry, but today the field of life science and specifically point of care is in the core of microfluidics. We have presented applications where microfluidic chips show great advantages compared to conventional systems. The importance of these applications was illustrated by showing research papers related to these applications. Some important applications of microfluidic chip were introduced here. Microfluidics covers a wide range of applications such as microreactors, bioprinting, fuel cells, and many more.
Expertises
References
- Seemann, R., Brinkmann, M., Pfohl, T. & Herminghaus, S. Droplet based microfluidics. Reports Prog. Phys. 75, (2012).
- Paquin, F., Rivnay, J., Salleo, A., Stingelin, N. & Silva, C. Droplet Control Technologies for Microfluidic High Throughput Screening (µHTS). Muhsincan Sesen,a Tuncay Alan,a and Adrian Neild∗a 10715–10722 (2017) doi:10.1039/b000000x.
- Galas, J. C., Bartolo, D. & Studer, V. Active connectors for microfluidic drops on demand. New J. Phys. 11, (2009).
- Jullien, M.-C., Tsang Mui Ching, M.-J., Cohen, C., Menetrier, L. & Tabeling, P. Droplet break in a low capillary T-junction. in 19th Mechanical French Congress (AFM, Maison de la Mécanique, 39/41 rue Louis Blanc-92400 Courbevoie, 2009).
- Yu, L., Chen, M. C. W. & Cheung, K. C. 2010 Droplet-based microfluidic system for multicellular tumor spheroid formation and anticancer drug testing. Lab Chip 10, 2424–2432 (2010).
- N.Shembekar, C.Chaipan, R. U. & C. A. M. 2016 Droplet-based microfluidics in drug discovery, transcriptomics and high-throughput molecular genetics. Lab Chip (2016) doi:10.1039/C6LC00249H.
- Shang, L., Cheng, Y. & Zhao, Y. Emerging Droplet Microfluidics. Chem. Rev. 117, 7964–8040 (2017).
- Suea-Ngam, A., Howes, P. D., Srisa-Art, M. & Demello, A. J. Droplet microfluidics: From proof-of-concept to real-world utility? Chem. Commun. 55, 9895–9903 (2019).
- Vladisavljević, G. T., Al Nuumani, R. & Nabavi, S. A. Microfluidic production of multiple emulsions. Micromachines 8, (2017).
- Soppimath, K. S. & Aminabhavi, T. M. Ethyl acetate as a dispersing solvent in the production of poly(DL-lactide-co-glycolide) microspheres: Effect of process parameters and polymer type. J. Microencapsul. 19, 281–292 (2002).
- Qi, F., Wu, J., Li, H. & Ma, G. Recent research and development of PLGA / PLA microspheres / nanoparticles : A review in scienti fi c and industrial aspects. (2018).
- Anderson, J. M. & Shive, M. S. Biodegradation and biocompatibility of PLA and PLGA microspheres. Adv. Drug Deliv. Rev. 64, 72–82 (2012).
- Macosko, E. Z. et al. Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell 161, 1202–1214 (2015).
- Halldorsson, S., Lucumi, E., Gómez-Sjöberg, R. & Fleming, R. M. T. Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices. Biosens. Bioelectron. 63, 218–231 (2015).
- Yeo, L. Y., Chang, H. C., Chan, P. P. Y. & Friend, J. R. Microfluidic devices for bioapplications. Small 7, 12–48 (2011).
- Coluccio, M. L. et al. Microfluidic platforms for cell cultures and investigations. Microelectron. Eng. 208, 14–28 (2019).
- Ayuso, J. M. et al. Development and characterization of a microfluidic model of the tumour microenvironment. Sci. Rep. 6, 1–16 (2016).
- Caballero, D. et al. Organ-on-chip models of cancer metastasis for future personalized medicine: From chip to the patient. Biomaterials 149, 98–115 (2017).
- Huh, D. et al. Reconstituting organ-level lung functions on a chip. Science (80-. ). 328, 1662–1668 (2010).
- Bhise, N. S. et al. Organ-on-a-chip platforms for studying drug delivery systems. J. Control. Release 190, 82–93 (2014).
- Zhang, B., Korolj, A., Lai, B. F. L. & Radisic, M. Advances in organ-on-a-chip engineering. Nat. Rev. Mater. 3, 257–278 (2018).
- C. Wyatt Shields IV, Dr. Catherine D. Reyes, and P. G. P. L. Microfluidic Cell Sorting: A Review of the Advances in the Separation of Cells from Debulking to Rare Cell Isolation. Lab Chip (2015) doi:10.1039/c4lc01246a.
- Kuntaegowdanahalli, S. S., Bhagat, A. A. S., Kumar, G. & Papautsky, I. Inertial microfluidics for continuous particle separation in spiral microchannels. Lab Chip 9, 2973–2980 (2009).
- Vivek, V., Zeng, Y. & Kim, E. S. NOVEL ACOUSTIC-WAVE MICROMIXER.
- Ward, K. & Fan, Z. H. Mixing in microfluidic devices and enhancement methods. J. Micromechanics Microengineering 25, (2015).
- Kim, H. et al. Submillisecond organic synthesis: Outpacing Fries rearrangement through microfluidic rapid mixing. Science (80-. ). 352, 691–694 (2016).
- Thorsen, T., Maerkl, S. J. & Quake, S. R. Microfluidic large-scale integration. Science (80-. ). 298, 580–584 (2002).
- Aditya Aryasomayajula, Pouriya Bayat, Pouya Rezai, P. R. S. Microfluidic Devices and Their Applications. Springer vol. 50 (2017).
- Jensen, E. C., Bhat, B. P. & Mathies, R. A. A digital microfluidic platform for the automation of quantitative biomolecular assays. Lab Chip 10, 685–691 (2010).
- Vincent, M., Xu, Y. & Kong, H. Helicase-dependent isothermal DNA amplification. EMBO Rep. 5, 795–800 (2004).
- Erik C. Jensen, Yong Zeng, Jungkyu Kim, and R. A. M. Microvalve Enabled Digital Microfluidic Systems for High Performance Biochemical and Genetic Analysis. 15, 455–463 (2011).
- Kim, J., Jensen, E. C., Stockton, A. M. & Mathies, R. A. Universal microfluidic automaton for autonomous sample processing: Application to the mars organic analyzer. Anal. Chem. 85, 7682–7688 (2013).
- Oh, K. W. & Ahn, C. H. A review of microvalves. J. Micromechanics Microengineering 16, (2006).
- Kim, J., Stockton, A. M., Jensen, E. C. & Mathies, R. A. Pneumatically actuated microvalve circuits for programmable automation of chemical and biochemical analysis. Lab Chip 16, 812–819 (2016).
- Chin, V. I. et al. Microfabricated platform for studying stem cell fates. Biotechnol. Bioeng. 88, 399–415 (2004).
- Yeo, J. C., Kenry & Lim, C. T. Emergence of microfluidic wearable technologies. Lab Chip 16, 4082–4090 (2016).
- Ahyeon Koh, Daeshik Kang, Yeguang Xue, Seungmin Lee, Rafal M. Pielak, Jeonghyun Kim, Taehwan Hwang, Seunghwan Min,1 Anthony Banks, Philippe Bastien, Megan C. Manco, Liang Wang, Kaitlyn R. Ammann, Kyung-In Jang, Phillip Won Seungyong Han, Roozbeh Ghaffari, J. A. R. A soft, wearable microfluidic device for the capture, storage, and colorimetric sensing of sweat. Sci. Transl. Med. 26, 39–46 (2017).
The Lamprou Lab (Queen’s University Belfast)
The Lamprou lab, affiliated with Queen’s University Belfast, specializes in three main areas: nanoparticles for imaging and therapy, lab-on-a-chip technology, and therapeutic implants. Their interdisciplinary approach has driven innovation in healthcare since 2012, developing emerging technologies and novel drug delivery devices. The lab is led by Professor Dimitrios Lamprou, a leading expert in pharmaceutical technologies known for his significant contributions to 3D printing, microfluidics, and nanofibers, with over 150 peer-reviewed publications.
What are engineered nanoparticles?
Nanotechnology gained prominence after Richard P. Feynman’s 1959 famous lecture, “There’s plenty of room at the bottom”.1 However, the use of nanoparticles can be traced back to ancient times, as seen in the Romans’ incorporation of nanoparticles into glass manufacturing in the fourth century AD. The Lycurgus cup, an artifact from this period, notably displayed distinctive color changes under various lighting conditions due to the integration of nano-glass particles.2
In modern times, nanotechnology has developed into a comprehensive scientific discipline with diverse applications spanning multiple industries. From water purification and information technologies to drug development, environmental solutions, and the creation of robust yet lightweight materials, nanotechnology has emerged as a pivotal player.3,4
The fundamental units of nanotechnology are nanoparticles, defined as small particles ranging from 10 nm to 1000 nm in size. Engineered nanoparticles, specifically designed with dimensions under 100 nm, play a crucial role in manipulating materials at the molecular and atomic levels, demonstrating significant chemical, structural, electrical, biological and mechanical characteristics. They are classified into categories including ceramic, carbon-based, semiconductor, metal, lipid-based and polymeric nanoparticles.
What makes engineered nanoparticles promising in nanomedicine?
Engineered nanoparticles hold promise for various nanotechnology applications in medicine, including in-vivo and in-vitro diagnosis, drug delivery, and production of biocompatible materials. They are characterized by their high mass-to-surface area ratio, ability to adsorb and carry compounds, and quantum properties.5,6
These characteristics can benefit the drug delivery field, as its primary objective is to enhance the specificity of drug targeting, improve safety and biocompatibility, and reduce toxicity while maintaining therapeutic effects. Nanoparticles with dimensions less than 100 nm are considered excellent drug carriers due to their unique biological and physiological properties, allowing them to cross tissue and cell barriers effectively.7
Engineered nanoparticles also exhibit a higher likelihood of cellular uptake due to their larger surface area, enabling increased protein loading. The interaction of NPs with biological substances results in the formation of a “protein corona,” enhancing nanocarriers’ uptake by the reticuloendothelial system. This protein corona can serve as a functional carrier for targeted drug delivery, improving poorly soluble drug uptake, directing drugs to specific locations, and enhancing drug bioavailability.8
Overall, engineered nanoparticle applications in nanomedicine have been progressing recently, particularly in controlled drug delivery, nucleic acid-based treatment, cancer cell targeting, angiogenesis inhibition, and inflammation control.
How are engineered nanoparticles traditionally produced
The success of nanoparticles as drug carriers and in nanomedicine applications depends on a number of crucial factors, including NP fabrication strategies, physical properties, drug loading efficiencies, drug release potential, and especially the carrier’s toxicity.
Lipid-based nanoparticles exhibit low toxicity in in-vivo experiments. They can carry both hydrophilic and hydrophobic molecules, leading to prolonged half-life and controlled drug release. Here, we focus on two main classes of lipid-based nanoparticles: liposomes and solid lipid nanoparticles.
Liposomes
A liposome is a microsphere lipid constructed from one or multiple phospholipid bilayers, closely mirroring the structure of cell membranes. The liposome preparation process consists of three main stages: preparing aqueous and lipid phases, primary processing with lipids, and optional secondary processing steps. Most methods involve dissolving phospholipids in an organic solvent, with subsequent removal of the solvent through evaporation—a critical step in liposome formation. Two straightforward methods of liposome synthesis are film hydration and solvent injection.
Liposomes generated through film hydration tend to be polydisperse. Parameters like the duration of rotary evaporation, mixing speed, and temperature after hydration affect liposome quality, emphasizing the need for careful monitoring.10,11
The major factors to consider for use of the solvent injection method are the temperature during injection and the injection rate. These factors will affect the size, shape, and polydispersity of the liposomes produced. The solvent injection method involves certain challenges, with continuous exposure of therapeutic substances to high temperatures and organic solvents affecting liposomal product stability and safety.
This results in high polydispersity and a non-homogeneous formulation.12
Solid Lipid Nanoparticles
Solid lipid nanoparticles (SLN) have emerged as a promising alternative to other lipid formulations like liposomes or polymeric nanoparticles. These spherical colloidal particles, ranging from 10 to 1000 nm in size, offer controlled drug release due to limited drug mobility in the solid lipid. SLNs provide advantages such as targeted drug delivery, controlled release, increased stability, scalability in production, and avoidance of organic solvents. Comprising a solid lipid core in an aqueous medium with a surfactant, SLNs use different lipids (steroids, fatty acids, triglycerides, etc.) and require stabilizing agents like surfactants or emulsifiers. The drug insertion process depends on drug hydrophobicity, solid lipid category, and polymeric alterations in the lipid. Production techniques include high-pressure homogenization, solvent evaporation, ultrasonication, hot homogenization, microemulsion, and others.13,14
Table: comparison of the different manufacturing methods used for polymeric NPs and lipid-based NPs
Nanoparticles Type | Manufacturing Method | Advantages | Disadvantages |
---|---|---|---|
Lipid formulation | Film hydration | – Established method – Understood method | – High consuming of the organic solvents – High PDI – Lack of reproducibility – Need for additional downsizing step – Difficulties in scaling-up |
Lipid formulation | Solvent injection | – Simple and fast – Scaling-up possibility | – Exposing to organic solvent – High PDI – Stability problems |
Lipid formulation | Extrusion | – Uniform and homogenous formulation | – Possible clogging of the membrane pores. – Difficulties in scaling-up |
Lipid formulation | High pressure homogenization | – Scaling-up possibility – Uniform formulation | – High energy consumption – Multiple steps – Bulky system |
Lipid formulation | Microemulsion | – Small particle size – Homogenous formulation | – Difficulty in removing the excess water – Use high concentration of surfactants |
Nanoparticles Type | Manufacturing Method | Advantages | Disadvantages |
---|---|---|---|
Polymeric | Emulsification-salting out | – Avoids surfactants and chlorinated solvents | – Need for purification steps – Encapsulate lipophilic drugs only |
Polymeric | Emulsification solvent diffusion | – Scaling-up possibility – Batch-to-batch reproducibility | – The possible diffusion of the hydrophilic drug into the aqueous phase – The need to eliminate high volume of aqueous phase from the colloidal dispersion |
Polymeric | Emulsification- evaporation | – Simple and versatile | – Risk of nanodroplets coalescence during the evaporation process – Time consuming |
Polymeric | Dialysis | – Effective and simple method – Produce polymeric nanoparticles with narrow distribution | – Time consuming – Use of high amount of dialyzing medium, which stimulate the premature release of NPs content |
Polymeric | Nonparticipation | – Simple and established method – Use low concentrations of surfactant | – Restricted for lipophilic drugs – Low polymer concentration obtained |
How does microfluidics enhance the properties of engineered nanoparticles
The principal innovation of microfluidics is the ability to transfer the traditional bulk technique to microscale fluidic chips. Solvents can be mixed within microchannels by a pumping system with continuous laminar flow. This type of flow offers high mixing quality and enhances the performance of microscale devices. The ability to adjust the flow rate ratio (FRR) and total flow rate (TFR) allows for continuous production of monodisperse and homogenous engineered nanoparticles.
Fluigent’s pressure-driven controllers and flow sensors enable this high stability and fast response in fluid flow, which is highly challenging to achieve with traditional pumping methods like peristaltic or syringe pumps. Overall, this method offers high reproducibility and low batch-to-batch variations. In addition, the method’s versatility makes the encapsulation process faster while keeping encapsulation efficiency high.
Microfluidics offers a fast, simple, single-step technique for liposome manufacturing, but challenges for large-scale production currently hinder its wide-scale implementation due to high costs.15,16
Most microfluidics-based solid lipid nanoparticle manufacturing follows a similar procedure involving dissolving lipids and drugs in an organic solvent, which is then introduced into the microfluidic device alongside an aqueous phase with a surfactant. Control over lipid-to-drug concentration, flow rate, and velocity influences the final nanocarrier characteristics.
Solid lipid nanoparticles produced by microfluidic processes have smaller particle sizes, better homogeneity, and higher encapsulation efficiency compared to bulk methods. In addition, cytotoxic studies demonstrate potent anti-proliferative effects of microfluidic SLNs in cancer cell lines.
However, research on microfluidic-produced SLNs is limited, and challenges exist, particularly with regard to the material used for microfluidic chips, with PDMS-based chips being sensitive to organic solutions.17
Conclusion
In this paper highlight, we presented the promise and challenges of microfluidics technology for the design and formulation of nanomedicines. While traditional methods face limitations in producing small engineered nanoparticles with desirable characteristics, microfluidic systems offer a one-step, controllable process with improved outcomes.
Comparative studies have shown that nanocarriers produced by microfluidics exhibit superior properties. Future advancements in microfluidic systems, combined with complementary tools like process analytical technology and molecular imaging technologies, are expected to optimize NP production and expand their medical applications.
Microfluidics holds promise for shaping the future of engineered nanoparticle research and development.
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References
(1) Shirai, Y.; Osgood, A.J.; Zhao, Y.; Yao, Y.; Saudan, L.; Yang, H.; Yu-Hung, C.; Alemany, L.B.; Sasaki, T.; Morin, J.-F. Surface-rolling molecules. J. Am. Chem. Soc. 2006, 128, 4854–4864.
(2) Bayda, S.; Adeel, M.; Tuccinardi, T.; Cordani, M.; Rizzolio, F. The History of Nanoscience and Nanotechnology: From Chemical-Physical Applications to Nanomedicine. Molecules 2019, 25, 112. Schoenmaker L, et al. mRNA-lipid nanoparticle COVID-19 vaccines: structure and stability. Int Pharm. 2021;601: 120586 .
(3) Grobert, N.; Hutton, D. Nanoscience and nanotechnologies: Opportunities and uncertainties. Lond. R. Soc. R. Acad. Eng. Rep.2004, 46, 618.
(4) Thiruvengadam, M.; Rajakumar, G.; Chung, I.M. Nanotechnology: Current uses and future applications in the food industry.3 Biotech 2018, 8, 74.
(5) Duncan, R. The dawning era of polymer therapeutics. Nat. Rev. Drug Discov. 2003, 2, 347–360.
(6) De Jong, W.H.; Borm, P.J. Drug delivery and nanoparticles:applications and hazards. Int. J. Nanomed. 2008, 3, 133–149.
(7) LaVan, D.A.; McGuire, T.; Langer, R. Small-scale systems for in vivo drug delivery. Nat. Biotechnol. 2003, 21, 1184–1191.
(8) Nel, A.E.; Mädler, L.; Velegol, D.; Xia, T.; Hoek, E.M.; Somasundaran, P.; Klaessig, F.; Castranova, V.; Thompson, M. Understanding biophysicochemical interactions at the nano-bio interface. Nat. Mater. 2009, 8, 543–557.
(9) Mitchell, M.J., Billingsley, M.M., Haley, R.M. et al. Engineering precision nanoparticles for drug delivery. Nat Rev Drug Discov 20, 101–124 (2021).
(10) Jaradat, E.; Weaver, E.; Meziane, A.; Lamprou, D. A. Microfluidics Technology for the Design and Formulation of Nanomedicines. Nanomaterials 2021, 11 (12), 3440.
(11) Alam, S.; Mattern-Schain, S.; Best, M. Targeting and triggered release using lipid-based supramolecular assemblies as medicinal nanocarriers. In Comprehensive Supramolecular Chemistry II; Elsevier: Oxford, UK, 2017; pp. 329–364.
(12) Maherani, B.; Arab-Tehrany, E.; Mozafari, M.R.; Gaiani, C.; Linder, M. Liposomes: A review of manufacturing techniques and targeting strategies. Curr. Nanosci. 2011, 7, 436–452.
(13) Mehnert, W.; Mäder, K. Solid lipid nanoparticles: Production, characterization and applications. Adv. Drug Deliv. Rev. 2001, 47,165–196.
(14) Uner, M.; Yener, G. Importance of solid lipid nanoparticles (SLN) in various administration routes and future perspectives. Int. J.Nanomed. 2007, 2, 289–300.
(15) Weaver, E.; Uddin, S.; Cole, D.K.; Hooker, A.; Lamprou, D.A. The Present and Future Role of Microfluidics for Protein and Peptide-Based Therapeutics and Diagnostics. Appl. Sci. 2021, 11, 4109.
(16) Guimarães Sá Correia, M.; Briuglia, M.L.; Niosi, F.; Lamprou, D.A. Microfluidic manufacturing of phospholipid nanoparticles: Stability, encapsulation efficacy, and drug release. Int. J. Pharm. 2017, 516, 91–99.
(17) Arduino, I.; Liu, Z.; Rahikkala, A.; Figueiredo, P.; Correia, A.; Cutrignelli, A.; Denora, N.; Santos, H.A. Preparation of cetylpalmitate-based PEGylated solid lipid nanoparticles by microfluidic technique. Acta Biomater. 2021, 121, 566–578.
Features of the cell encapsulation platform for flow cytometry
➤ Fluigent Precision and Flexibility
Produce robust and highly monodisperse emulsions using Fluigent’s pressure-based flow controllers and the Raydrop while accurately controlling the size of your droplets and shell thickness.
➤ Start emulsions production right away
The system is a fully equipped, mounted and controlled tool to create double emulsions with little setup time.
➤ Complete and easy-to-use engineered system
Our organized system requires easy priming and cleaning processes for better performance. It includes dedicated optics for optimized droplet visualization at a high frequency.
➤ Innovative and much sought-after application for cell analysis
The system is an easy-to-use platform for creating single-cell encapsulations into double emulsion droplets compatible with high-throughput screening and FACS experiments.
The encapsulation platform for FACS allows:
- Elimination of the risk of cross-contamination.
- Fast and efficient mixing of the reagents that occurs inside droplets.
- Ability to work with cells of limited availability.
Description
The cell encapsulation platform includes an organized flow path with pressure controllers, filters, flowmeters, and valves to facilitate the start-up, shut-down, and cleaning of the system between runs. Dedicated optics are included to optimize the visualization of droplet production at high generation frequency.
What is FACS technology?
In the field of biology, FACS stands for Fluorescence-Activated Cell Sorting. It is a technology aimed at separating and isolating certain cells from a heterogeneous population based on their fluorescent properties. This fluorescent signal allows users to translate the production of certain molecules of interest or the expression of specific phenotypes. The process of this technique is to label the cells with fluorescent markers such as antibodies or dyes which have the role of binding to certain structures or molecules of the cell in a specific way. These labeled cells can then be passed through the flow cytometer, an instrument allowing for the detection and measurement of the fluorescence emitted by the cells. This fluorescence detection will allow the flow cytometer to sort the cells into different populations, allowing the researcher to collect and analyze these different groups of cells separately.
Using FACS enables the scientist to perform an efficient sorting with a limited number of doublets or multiple cells being sorted together and from there, the characterization of cellular heterogenicity and the rare cell subpopulation.
Through our microfluidic technology, the encapsulation platform for FACS allows for the generation of samples, which can be processed in flow cytometry.
How to use the platform for FACS experiments?
Microfluidics can be associated with FACS technology to improve results. Working with microliter volumes allows for a considerable reduction in cost since the volume of samples required is greatly reduced. For instance, it minimizes the need for large cell cultures.
In addition, the use of encapsulation methods in droplets allows users to confine the fluorescence signal, avoiding the use of chemicals, and to perform bioassays at the single cell scale. Single cell or bacteria growth in drops can also be considered.
As the encapsulation platform for droplet sorting produces double emulsions to encapsulate cells (from 5 µm to 40 µm), the limitations of the FACS are overcome. The first limitation is that the secreted molecules are generally detectable only if a binding protocol has been performed. In the opposite case, they remain dissolved in the surrounding medium. The second is that the secreted cells significantly impact each other. Combining droplet microfluidics and FACS technology then appears to be a major advance for research and allows users to obtain more reliable and reproducible data.
Our selection of webinars on double emulsion for FACS
Encapsulation of fluorescent bacteria in double emulsions for FACS
Learn how to use our double emulsion platform to encapsulate bacteria in W/O/W DE and discover the DE sorting efficiency using FACS.
Webinar summary:
- Introduction to FACS sorting: technology, advantages, and drawbacks
- Presentation of the platform: encapsulation of single bacteria in DE
- Microscopy characterization and FACS sorting
- Interactive Q&A session
Single cell encapsulations compatible with FACS sorting, API encapsulations in biocompatible polymers, and more.
Until now, performing high efficiency encapsulation processes continuously has been challenging, as existing technologies have limitations (low reproducibility, the need for multiple surface coatings, poor size distribution, etc). The Raydrop encapsulation platform from Secoya solves these issues by providing an easy-to-use system that produces highly monodisperse W/O/W or O/W/O single or double emulsion from 15 µm to 400 µm.
Webinar Summary:
- Theoretical presentation of the Raydrop’s encapsulation process
- Case study: encapsulation of cells, yeast, and bacteria in a small double emulsion for FACS sorting
- Case study encapsulation of API in microbeads and microcapsules
- Q&A
Cell encapsulation platform: Examples of use
Standard cell encapsulation platform
The standard cell encapsulation platform allows for the encapsulation of single cells for high throughput screening applications involving drug discovery, toxicity testing, various ‘omics’ studies and rare cell analysis1,2.
How to inject a small sample?
The encapsulation platform for flow cytometry with an injection loop is a tool capable of greatly improving working conditions as it handles fluid volumes, as low as a few dozen microliters. Adding the L-Switch is advantageous in many scientific and medical applications, making it possible to work with rarer cells.
For instance, researchers working on stem cells are often challenged by their low availability. Indeed, because of their limited distribution in time (available only during a period of human life, difficulties to self-renew into stem cells after differentiation) and space (often nested in environments favorable to their functioning, protection and regulation, which limit their dispersion in the organism) and their delicate and expensive conditions of culture, their large-scale production is not possible. Thus, to be able to work with small volumes of culture and thus a low number of cells is advantageous.
Another example is the use of primary cells, directly collected from the patient and useful for personalized medicine. Acquiring this type of cells can be logistically challenging and the available volume can also be limited. The cell encapsulation platform’s injection loop solves this predicament by enabling researchers to work with small volumes of patient-derived cells.
Finally, using this platform for FACS also allows users to limit the use of expensive reagents and to limit wastes, unlocking new possibilities in the field of biomedical research and personalized medicine.
How to add a reagent?
In many microfluidic procedures, an integrated tooling for mixing is essential for appropriate functionality in a broad range of applications.
The usual encapsulation platform for drop allowing users to integrate a new phase for optimal agent mixing. It is recommended to mix two solutions in a fluidic path before encapsulation. After the precise formation of self‐ assembled lipid or polymeric particles by microfluidic mixing, the encapsulation of active molecules is loaded into synthesized particles.
The efficiency of many biosensors and the possibility of investigating reaction kinetics with quick time resolution depends on controlled mixing. It can be also used to encapsulate cells with barcoded beads for Next generation sequencing (DropSeq), for enzymatic tests, or phenotyping5.
Which applications can use the encapsulation platform for FACS?
High-throughput screening: Cell sorting
Cell encapsulation can improve high-throughput screening outcomes since the highly monodisperse droplets provide homogeneous reaction conditions. It can be combined with cell sorting tools such as Fluorescence Activated Cell Sorting (FACS), to achieve higher speed and efficiency compared to conventional approaches. Benefits include:
- Lower costs while increasing sequencing accuracy and depth:
- Sorting and sequencing only cells containing droplets could vastly reduce reagent costs and increase accuracy and depth for downstream Next Generation Sequencing, including eliminating common issues with single-cell droplet sequencing such as reads from empty droplets due to the encapsulation of free-floating transcripts.
- Increased range of measurement:
- Encapsulated cells may be sorted based on phenotypes not currently measurable with standard cell sorting, such as enzymatic turnover, presence of secreted molecules, or quantification of proteins lacking cell surface markers.
- Multi-omics profiling:
- In contrast with conventional methods, cell sorting using microfluidics provides a unique tool to link genotypes with phenotypes through compartmentalization.
- Our cell encapsulation platform allows for the combination of single-cell phenotyping and genome-wide sequencing, enabling multiomic measurements and directly linking cellular phenotypes to their underlying genetic mechanism2.
Drug delivery
Cell or drug encapsulation for drug delivery consists of the immobilization of bioactive materials, mainly cells, within a double emulsion, generally surrounded by a polymeric membrane. The latter permits the free passage of nutrients and oxygen and the egress of therapeutic protein products. Encapsulation allows for the protection of the cell content from mechanical stress and in the case of allogeneic tissue also from the host’s immune response. Microcapsules are commonly used in pharmaceutical or medical processes as drug carriers or for the encapsulation of organic cells.
Locally-implanted droplets may represent a minimally invasive manner of delivering therapeutic proteins such as growth factors and cytokines. It could also allow for mosaic injections containing several droplets to provide complex release profiles or multi-protein deliveries3.
Transplants
The microencapsulation of living cells may serve as an alternative therapy for patients requiring organ transplants. Researchers can control droplets size to mitigate the immune infiltration of transplanted cells while keeping oxygen and waste products transported. Droplets can also immobilize cells at the desired transplant size, which is essential as many cell therapies rely on systemic cell administration.
Cell encapsulation for cell-based therapy is emerging as a promising strategy for treating a wide range of human diseases, such as diabetes, blood disorders, acute liver failure, spinal cord injury, and several types of cancer through transplants. Pancreatic islets, blood cells, hepatocytes, and stem cells are among the many cell types currently used for this strategy. The encapsulation of these “therapeutic” cells is intended to prevent immune rejection, to provide a controlled and supportive environment, and to enable a complete retrieval of the graft in the case of an adverse body reaction4.
Specifications
We propose 4 different platforms to target different droplet sizes.
Description | Product | P.N |
Cell encapsulation device | Cell encapsulation platform | O-FACS1-PTF |
Double Emulsion production device | – RayDrop Double Emulsion 30-70-45 | – O-DE-RDRPC05- EUP |
Fluid handling system | – LINK Module – Flow EZ 7 bars for all 3 phases | – LU-LNK-0002 – LU-FEZ-7000PCK |
Reservoirs | – Continuous phase: 50 mL Pcap with 50 mL Falcon tube Tubing: 500 µm – Shell phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm – Core phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm | – P-CAP50-HP-PCK – P-CAP15-HP-PCK – P-CAP15-HP-PCK |
Flow Meters | – Continuous phase: Flow Unit L – Shell phase: Flow Unit M – Core phase: Flow Unit M | – FLU-L-D – FLU-M-D – FLU-M-D |
Optical System | – Light source – Microscope objective (x10) – Specific colour camera (up to 400 fps, 1µs integration time) – XYZ translation stages | N/A |
Tubing & Fittings | – Tubing: OD: 1/16 and 1/32 OD ID: 250 µm & 500 µm Materials: PFA – Manual valves: 4 way valves 2 way valves – Filters: 2 µm filter for continuous phase 2 µm filter for dispersed phase | N/A |
Wetted materials | – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS – Sealing: FFKM | N/A |
Unit dimensions | – 61 x 46 x 43cm 3 (L x W x H) | N/A |
Weight | – 4 kg without the protective wood – 21 kg with the protective wood | N/A |
Droplet Size Range* | 25-45 µm | N/A |
Description | Product | P.N |
Cell encapsulation device | Cell encapsulation platform | O-FACS2-PTF |
Double Emulsion production device | – RayDrop Double Emulsion 30-70-60 | – O-DE-RDRPC06- EUP |
Fluid handling system | – LINK Module – Flow EZ 7 bars for all 3 phases | – LU-LNK-0002 – LU-FEZ-7000PCK |
Reservoirs | – Continuous phase: 50 mL Pcap with 50 mL Falcon tube Tubing: 500 µm – Shell phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm – Core phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm | – P-CAP50-HP-PCK – P-CAP15-HP-PCK – P-CAP15-HP-PCK |
Flow Meters | – Continuous phase: Flow Unit L – Shell phase: Flow Unit M – Core phase: Flow Unit M | – FLU-L-D – FLU-M-D – FLU-M-D |
Optical System | – Light source – Microscope objective (x10) – Specific colour camera (up to 400 fps, 1µs integration time) – XYZ translation stages | N/A |
Tubing & Fittings | – Tubing: OD: 1/16 and 1/32 OD ID: 250 µm & 500 µm Materials: PFA – Manual valves: 4 way valves 2 way valves – Filters: 2 µm filter for continuous phase 2 µm filter for dispersed phase | N/A |
Wetted materials | – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS – Sealing: FFKM | N/A |
Unit dimensions | – 61 x 46 x 43cm 3 (L x W x H) | N/A |
Weight | – 4 kg without the protective wood – 21 kg with the protective wood | N/A |
Droplet Size Range | 45-60 µm | N/A |
Description | Product | P.N |
Cell encapsulation device | Cell encapsulation platform | O-FACS3-PTF |
Double Emulsion production device | – RayDrop Double Emulsion 60-120-60 | – O-DE-RDRPC07- EUP |
Fluid handling system | – LINK Module – Flow EZ 7 bars for all 3 phases | – LU-LNK-0002 – LU-FEZ-7000PCK |
Reservoirs | – Continuous phase: 50 mL Pcap with 50 mL Falcon tube Tubing: 500 µm – Shell phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm – Core phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm | – P-CAP50-HP-PCK – P-CAP15-HP-PCK – P-CAP15-HP-PCK |
Flow Meters | – Continuous phase: Flow Unit L – Shell phase: Flow Unit M – Core phase: Flow Unit M | – FLU-L-D – FLU-M-D – FLU-M-D |
Optical System | – Light source – Microscope objective (x5) – Specific colour camera (up to 400 fps, 1µs integration time) – XYZ translation stages | N/A |
Tubing & Fittings | – Tubing: OD: 1/16 and 1/32 OD ID: 250 µm & 500 µm Materials: PFA – Manual valves: 4 way valves 2 way valves – Filters: 2 µm filter for continuous phase 2 µm filter for dispersed phase | N/A |
Wetted materials | – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS – Sealing: FFKM | N/A |
Unit dimensions | – 61 x 46 x 43cm 3 (L x W x H) | N/A |
Weight | – 4 kg without the protective wood – 21 kg with the protective wood | N/A |
Droplet Size Range | 50-60 µm | N/A |
Description | Product | P.N |
Cell encapsulation device | Cell encapsulation platform | O-FACS4-PTF |
Double Emulsion production device | – RayDrop Double Emulsion 60-120-90 | – O-DE-RDRPC08- EUP |
Fluid handling system | – LINK Module – Flow EZ 7 bars for all 3 phases | – LU-LNK-0002 – LU-FEZ-7000PCK |
Reservoirs | – Continuous phase: 50 mL Pcap with 50 mL Falcon tube Tubing: 500 µm – Shell phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm – Core phase: 15 mL Pcap with 15 mL Falcon tube Tubing: 125 µm | – P-CAP50-HP-PCK – P-CAP15-HP-PCK – P-CAP15-HP-PCK |
Flow Meters | – Continuous phase: Flow Unit L – Shell phase: Flow Unit M – Core phase: Flow Unit M | – FLU-L-D – FLU-M-D – FLU-M-D |
Optical System | – Light source – Microscope objective (x5) – Specific colour camera (up to 400 fps, 1µs integration time) – XYZ translation stages | N/A |
Tubing & Fittings | – Tubing: OD: 1/16 and 1/32 OD ID: 250 µm & 500 µm Materials: PFA – Manual valves: 4 way valves 2 way valves – Filters: 2 µm filter for continuous phase 2 µm filter for dispersed phase | N/A |
Wetted materials | – Platform: PEEK, PFA, PCTFE, PTFE, SS316L, GLASS – Sealing: FFKM | N/A |
Unit dimensions | – 61 x 46 x 43cm 3 (L x W x H) | N/A |
Weight | – 4 kg without the protective wood – 21 kg with the protective wood | N/A |
Droplet Size Range | 70-90 µm | N/A |
OxyGEN
Control in real-time, protocol automation, data record and export |
ver. 2.2.0.0 or more recent |
Software Development Kit
Custom software application |
ver. 22.2.0.0 or more recent |
References
- Steele, J.A.M. et al. (2014) “Therapeutic cell encapsulation techniques and applications in diabetes,” Advanced Drug Delivery Reviews, 67-68, pp. 74–83. Available at: https://doi.org/10.1016/j.addr.2013.09.015.
Collins, D.J. et al. (2015) “The Poisson distribution and beyond: Methods for microfluidic droplet production and single cell encapsulation,” Lab on a Chip, 15(17), pp. 3439–3459. Available at: https://doi.org/10.1039/c5lc00614g.
Headen, D.M., García, J.R. and García, A.J. (2018) “Parallel droplet microfluidics for high throughput cell encapsulation and synthetic microgel generation,” Microsystems & Nanoengineering, 4(1). Available at: https://doi.org/10.1038/micronano.2017.76.
Farina, M. et al. (2019) “Cell encapsulation: Overcoming barriers in cell transplantation in diabetes and beyond,” Advanced Drug Delivery Reviews, 139, pp. 92–115. Available at: https://doi.org/10.1016/j.addr.2018.04.018.
- Damiati, S. et al. (2018) “Microfluidic devices for drug delivery systems and drug screening,” Genes, 9(2), p. 103. Available at: https://doi.org/10.3390/genes9020103.
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Expertise & resources
Introduction
Despite considerable progress in recent years, various disease diagnostics and treatments continue to present constraints such as low sensitivity or specificity, drug toxicity, and severe side effects [1]. Cancer represents one of the best examples of a disease where localized delivery of therapeutics is of great importance, as the potent yet toxic mechanisms of action of such compounds can lead to either an effective response or side effects. Today, most drug formulations are not capable of targeting specific sites of interest. Nanoparticle-based drug delivery platforms have emerged as suitable vehicles for overcoming these limitations [2]. Nanoparticles, such as liposomes, have proven advantageous at preserving therapeutic material and allowing for extended half-lives of drugs within the body [3].
Liposome nanoparticles are specialized delivery vehicles that serve multiple roles in enhancing the capabilities of active pharmaceutical ingredients (APIs). They can shield a drug from detection by the body’s immune system, and they serve to help solubilize highly lipophilic drug molecules or modulate the pharmacokinetics and biodistribution of the API.
What are liposome nanoparticles?
Liposome nanoparticles were discovered in the 1960s. These hollow nanoparticles are phospholipid vesicles consisting of at least one lipid bilayer (figure 1). This bilayer is usually composed of amphiphilic phospholipids that have a hydrophilic phosphate head and a hydrophobic tail consisting of two fatty acid chains. This structural feature has facilitated liposome applications, including their use as artificial cell membranes, carriers for drug delivery systems, encapsulating agents for food ingredients, and analytical tools [4–8].
Recent state-of-the-art applications
During the COVID-19 pandemic, the first vaccines to reach clinical trials were based on viral vector and nucleic acid technologies. One of the most promising vaccine candidates was based on nucleoside-modified mRNA and encapsulated within lipid nanoparticles (LNP) [9]. This confirms the need for lipidic nanoparticles for present and future drug delivery applications.
In recent years, liposome nanoparticles have attracted significant attention as a trusted class of drug delivery vehicles. Their self-closed structures can encapsulate multiple drugs at once, protecting the enclosed cargo from hydrolysis and breakdown. Additionally, targeting proteins and surface functional ligands on the outer shell of the lipid bilayer can add novel functionality—enabling targeted entry of liposomes into cells, either via antibodies or receptor-targeted ligands. These ligands attach to cell receptors that are over-expressed in certain diseased cells, allowing entry of the drug through the cell membrane.
What is the difference between Liposome and Lipid Nanoparticles?
Liposome and lipid nanoparticles are lipid-based structures used for drug delivery. Liposomes are spherical vesicles with lipid bilayers surrounding an aqueous core, while lipid nanoparticles are solid particles composed of solid lipids or a mixture of solid and liquid lipids. Liposomes have a hydrophilic outer layer and can encapsulate both hydrophilic and hydrophobic molecules, offering versatility but potentially reduced stability.
In contrast, lipid nanoparticles provide improved stability and are relatively easier to manufacture at scale. They are especially suitable for encapsulating poorly water-soluble drugs or nucleic acids. Liposomes have been widely used, while lipid nanoparticles are gaining in popularity, particularly in mRNA-based vaccines like those for COVID-19. Both lipid-based systems contribute to advanced drug delivery strategies, with liposomes offering flexibility and lipid nanoparticles providing enhanced stability and ease of production.
Comparison with Another Production Method
Batch method | Fluigent microfluidic method | |
Particle size distribution | Low | High |
Reproducibility | Low | High |
Live particle size control | No | Precise |
Range of particle size | Limited size range | Wide size range |
Continuous (/in line) production | No | Yes |
How to produce liposome nanoparticles
Equipment
Liposome nanoparticle generation
A stream of lipid in alcohol solution is surrounded by an aqueous phase within a glass capillary. The alcohol solution containing lipids diffuses into the aqueous solution (and reciprocally the water diffuses into the alcohol), until the alcohol concentration decreases to the solubility limit of the lipids. Consequently, this diffusion triggers the formation of liposomes by a mechanism described as “self-assembly”, where lipids assemble into a more energetically favorable structure.
The liposome nanoparticle production system is illustrated in figure 2.
Partial results
Conclusion
Liposome nanoparticles have proven advantageous for solubilizing therapeutic substances. Macroscale batch methods widely employed for liposome production lack control over liposome morphology, size, and distribution. Microfluidic systems allow for the production of highly monodisperse liposome nanoparticles. We have demonstrated the production of liposomes using a microfluidic system consisting of pressure-based flow controllers and the Raydrop™ microfluidic device with standard configuration. Liposomes ranging from 30 to 150 nm were generated. Sizes can be adjusted by controlling the device flow input parameters, particularly the flow rate ratio (FRR). The polydispersity index (PDI) ranges from 0.07 to 0.15. This system enables synthesis of liposomes for drug delivery applications, as encapsulating agents for food ingredients, or for other applications requiring nano-sized and spherical liposomes.
A complete, cost-effective and commercially-available platform for on-demand production of monodisperse liposome nanoparticles is now available. This allows for control of liposome size and frequency by adjusting flow parameters.
Related Resources
- 논문 하이라이트
Microfluidic technology for engineered nanoparticles in nanomedicine
Read more - 사용 설명서
Liposome nanoparticles production station Protocol
Download - 기술 데이터시트
Liposome nanoparticles production station Datasheet
Download - 사용 설명서
Nanoparticle Production Station User Manual
Download - 기술 데이터시트
PLGA Nanoparticle Production Station Datasheet
Download - 전문성 검토
Microfluidics for vaccine development
Read more - 기술서
Droplet-based Microfluidics
Read more
References
- Bozzuto, G. & Molinari, A. Liposomes as nanomedical devices. Int. J. Nanomedicine 10, 975–999 (2015).
- Blanco, E., Shen, H. & Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 33, 941–951 (2015).
- Torchilin, V. P. Recent advances with liposomes as pharmaceutical carriers. Nat. Rev. Drug Discov. 4, 145–160 (2005).
- Bally, M. et al. Liposome and lipid bilayer arrays towards biosensing applications. Small 6, 2481–2497 (2010).
- Fathi, M., Mozafari, M. R. & Mohebbi, M. Nanoencapsulation of food ingredients using lipid based delivery systems. Trends Food Sci. Technol. 23, 13–27 (2012).
- Grimaldi, N. et al. Lipid-based nanovesicles for nanomedicine. Chem. Soc. Rev. 45, 6520–6545 (2016).
- Andrew Pohorille & David Deamer. Artificial cells: prospects for biotechnology. Trends Biotechnol. Biotechnol. 31- (2002).
- Rongen, H. A. H., Bult, A. & Van Bennekom, W. P. Liposomes and immunoassays. J. Immunol. Methods 204, 105–133 (1997).
- Vogel, A. B. et al. A prefusion SARS-CoV-2 spike RNA vaccine is highly immunogenic and prevents lung infection in non-human primates. bioRxiv 2020.09.08.280818 (2020).
- Pattni, B. S., Chupin, V. V. & Torchilin, V. P. New Developments in Liposomal Drug Delivery. Chem. Rev. 115, 10938–10966 (2015).
- Mui, B., Chow, L. & Hope, M. J. Extrusion Technique to Generate Liposomes of Defined Size. Methods Enzymol. 367, 3–14 (2003).
- Carugo, D., Bottaro, E., Owen, J., Stride, E. & Nastruzzi, C. Liposome production by microfluidics: Potential and limiting factors. Sci. Rep. 6, 1–15 (2016).
- Hood, R. R., Devoe, D. L., Atencia, J., Vreeland, W. N. & Omiatek, D. M. A facile route to the synthesis of monodisperse nanoscale liposomes using 3D microfluidic hydrodynamic focusing in a concentric capillary array. Lab Chip 14, 2403–2409 (2014).
- Jahn, A., Vreeland, W. N., Devoe, D. L., Locascio, L. E. & Gaitan, M. Microfluidic directed formation of liposomes of controlled size. Langmuir 23, 6289–6293 (2007).
- Jahn, A. et al. Microfluidic mixing and the formation of nanoscale lipid vesicles. ACS Nano 4, 2077–2087 (2010).
Microfluidics in Pharmaceutics
Drug delivery
Microparticles and emulsions are used for a wide variety of pharmaceutical products including intravenous, intramuscular, ocular, or orally delivered compounds. Emulsions are also used as templates for polymer microparticles, lipid nanoparticles, or microcapsules. These are later used for drug delivery, with the emulsion being the active pharmaceutical ingredient (API) itself, or as an adjuvant for co-administration.
Droplet microfluidics technology produces multiunit drug delivery systems with precise dosage control, targeted release, and homogeneous distribution. Conventional methods struggle to achieve monodispersity (<5%) required for efficient drug delivery. Our technology revolutionizes drug delivery, offering improved treatment options and personalized therapies and advancing pharmaceutical development and patient care.
To learn more about droplet microfluidics for pharmaceutical applications, read our white paper about droplet microfluidics.
Disease modeling and characterization
Animal studies are the current standard for evaluating potential treatments, but they often struggle to correlate with human outcomes due to physiological differences and genetic variations. These studies are costly, time-consuming, and raise ethical concerns. Human clinical studies face challenges due to individual diversity and research complexity. Microfluidic models in an organ-on-chip format offer a promising alternative, providing an efficient, cost-effective, and ethical approach for drug discovery and personalized medicine. This technology advances the field of pharmaceutics by enabling the development of predictive methods to evaluate new compounds and therapeutics. It then helps in disease modeling and characterization, enhancing the drug discovery and development process.
To delve deeper into the topic of microfluidics for pharmaceutical applications using organ-on-chip systems, read our OOC white paper.
Industrial applications
Research applications
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Industrial field
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From the life sciences to the food industry, many applications require the use of fluids driven at flow rates from nanoliters to milliliters per minute. At such low flows, the success of these applications strongly depends on the level of control and automation of the fluidic operations.
These applications require flow control systems that are adapted for ensuring their success.
What’s included in the pack
- Fluigent’s MFCS-EX: A microfluidic flow controller with 8 customizable channels with different pressure ranges for high precision operations in microfluidic experiments.
- Fluigent’s Flow Unit : Bidirectional microfluidic flow sensors available in multiple low-rate ranges – up to 8 flow units can be used in one platform.
- Incubator grid: An on-demand, customizable grid compatible with the incubator along with reservoir holders.
- Fluigent connectors and tubing kit.
Features of the cell perfusion system
Stable & Complex Flow Patterns
Fluigent products have unprecedented performances in terms of precise control, stability,and responsiveness. High stability is particularly useful for applications with constant shear stress, like vascular perfusion.
Our Cell Perfusion Pack creates high responsiveness in the reproduction of complex flow patterns such as aortic pressure variations. This level of control ensures consistent and reproducible experimental conditions, minimizing experimental variability.
Protocol automation & user-friendly interface
Once the parameters are set and optimized, protocol automation is the key to saving time, limiting contamination, and decreasing variability. Using any protocol, valves, or pressure, Fluigent flow controllers can be assembled to automate protocols using user-friendly software (OxyGEN).
Researchers can quickly become proficient in using the system, reducing the learning curve, and maximizing productivity.
Versatility
This package can be used for diverse applications with any microfluidic chip.
Enhanced High-Throughput Capabilities
With the ability to run multiple assays in parallel, our perfusion pack is specifically designed to accommodate high-throughput experimentation while efficiently generating data. Compatible with Various Applications
The package is versatile and suitable for a wide range of applications, including drug screening, disease modeling, toxicity testing, and personalized medicine studies.
Customization
Our high throughput perfusion pack is flexible and adaptable to various organ-on-a-chip models and research needs (specific flow rates, pressure, etc.) The cell perfusion package’s modularity allows researchers to design experiments tailored to their unique research questions.
Microfluidic perfusion for cell culture
Microfluidictechnology has revolutionized cell culture systems by enabling precise control over fluid flow and creating microenvironments that mimic physiological conditions. One key technique in microfluidics is cell perfusion, which involves the controlled flow of fluid over cells or tissues to maintain their viability and functionality.
Cell perfusion begins with the design of a microfluidic device that incorporates intricate channels or networks, typically on a micrometer scale. Cells are then seeded onto the device, either as a monolayer or in 3D structures, and a continuous flow of culture media or other desired fluids is established within the device. This flow is carefully regulated by a pressure-based controller to provide nutrients, remove waste products, and maintain a stable microenvironment.
The ability to precisely control the flow rate and duration of perfusion allows researchers to mimic physiological conditions, such as blood flow rates in blood vessels. Real-time monitoring and analysis of parameters like cell viability, proliferation, and metabolic activity provide valuable insights into cellular behavior within these microenvironments.
What are possible applications for our cell perfusion pack?
Drug Screening and Development
This setup facilitates the testing of potential drug candidates under more realistic physiological conditions. High-throughput capabilities enable the screening of numerous drug compounds simultaneously, accelerating the drug discovery process and reducing costs.
In this example of the application, Chakrabarty et al. (1) developed a novel microfluidic Cancer-on-a-Chip platform to evaluate patient treatment responses using controlled growth conditions for tumor tissue slices. This system allows for the effective prediction of treatment responses for breast and prostate tumor models, and the culture period could be extended up to 14 days without significant changes in tissue quality.
Disease Modeling and Pathophysiology Studies
By investigating cellular responses to disease-related factors, this cell perfusion pack could provide valuable insights into disease pathophysiology, facilitating the development of targeted therapies.
In this example, Messelmani et al. (2) developed a new liver-on-a-chip model integrating a hydroscaffold, allowing cells to organize into complex 3D spheroid architecture. This model could play a role in producing a promising device for disease modeling, drug screening,and risk assessment.
Another example of disease modeling using Fluigent’s MFCS pack is the work of Paloschi et al. (3). They developed an Artery-on-a-Chip model mimicking the arterial vessel wall by incorporating structural aspects of the vasculature (lumen-intima-media) as well as hemodynamic forces impacting the luminal cells. They were able to characterize novel targets previously unrelated to vascular diseases, and to demonstrate that this model system could be used as a platform for testing novel therapeutic agents.
Toxicity Testing and Safety Assessment
High-throughput capabilities of our high throughput cell perfusion pack enable researchers to screen multiple compounds for potential toxicity, helping identify safe and effective substances for further development.
Tissue Engineering and Regenerative Medicine
Researchers can employ the cell perfusion pack to create dynamic microenvironments that promote tissue growth and regeneration.
The platform enables the study of tissue development, cell behavior, and the interactions between different cell types, aiding tissue engineering efforts.
Understanding how tissues respond to different growth factors and conditions can enhance regenerative medicine approaches and tissue transplantation strategies.
Specifications
Fluigent package
Fluigent’s MFCS-EX (up to 8 channels) |
Fluigent’s Flow Unit (up to 8 flow unit packed in one platform) |
Incubator grid (On demand custom grid with reservoir holders) |
BEOnChip microfluidic chip |
Fluigent connectors and tubing kit |
OxyGEN
Control in real-time, protocol automation, data record and export |
ver. 2.2.0.0 or more recent |
Software Development Kit
Custom software application |
ver. 22.2.0.0 or more recent |
Expertise & resources
Related products
Accessories
References:
- Chakrabarty S, Quiros-Solano WF, Kuijten MMP, Haspels B, Mallya S, Lo CSY, Othman A, Silvestri C, van de Stolpe A, Gaio N, Odijk H, van de Ven M, de Ridder CMA, van Weerden WM, Jonkers J, Dekker R, Taneja N, Kanaar R, van Gent DC. A Microfluidic Cancer-on-Chip Platform Predicts Drug Response Using Organotypic Tumor Slice Culture. Cancer Res. 2022 Feb 1;82(3):510-520. doi: 10.1158/0008-5472.CAN-21-0799. Epub 2021 Dec 6. PMID: 34872965; PMCID: PMC9397621.
- Messelmani T, Le Goff A, Souguir Z, Maes V, Roudaut M, Vandenhaute E, Maubon N, Legallais C, Leclerc E, Jellali R. Development of Liver-on-Chip Integrating a Hydroscaffold Mimicking the Liver’s Extracellular Matrix. Bioengineering (Basel). 2022 Sep 5;9(9):443. doi: 10.3390/bioengineering9090443. PMID: 36134989; PMCID: PMC9495334.
- Utilization of an Artery-on-a-chip to unravel novel regulators 2 and therapeutic targets in vascular diseases 3 4 Valentina Paloschi1,2*, Jessica Pauli1,2, Greg Winski3 , Zhiyuan Wu1,4, Zhaolong Li1 5 , Nadiya Glukha1 , Nora Hummel1 , Felix Rogowitz5 , Sandro Meucci6 , Lorenzo Botti7 , Albert Busch1,8 6 , Ekaterina Chernogubova4 , Hong Jin4 , Nadja Sachs1 , Hans-Henning Eckstein1 7 , Reinier A. Boon9,10,11, Andreas R. Bausch12, Lars Maegdefessel1,2,3 https://www.biorxiv.org/content/10.1101/2022.11.29.517312v1
MIMLIVER ON CHIP
MIMLIVER on CHIP project coordinated by the laboratory of Biomechanics and Bioengineering from UTC/CNRS aims at developing a functional liver-on-chip system to evaluate the toxicity of drugs. It is motivated by the established observation that 90% of potential drug candidates fail in clinical trials due to the lack of relevant models in the early drug development stages. The Liver is a central organ for toxicology assessment as it metabolizes the drugs into compounds that can be more active than the parent drug itself. Prior to being metabolized by hepatocytes, drugs are first filtered across an endothelial barrier which is typical of the liver. This selective barrier loses its properties in pathological conditions.
MIMLIVER on CHIP project Kick Off
- October 18, 2019
Fluigent and our collaborators are pleased to announce that the 1.5 M€ project: MIMLIVER on CHIP has been funded by the French National Research Agency.
This project coordinated by the laboratory of Biomechanics and Bioengineering from UTC/CNRS aims at developing a functional liver-on-chip system to evaluate the toxicity of drugs.
This project is motivated by the established observation that 90% of potential drug candidates fail in clinical trials due to the lack of relevant models in the early drug development stages. The Liver is a central organ for toxicology assessment as it metabolizes the drugs into compounds that can be more active than the parent drug itself. Prior to being metabolized by hepatocytes, drugs are first filtered across an endothelial barrier which is typical of the liver. This selective barrier loses its properties in pathological conditions.
In order to produce more reliable results in preclinical development, the MIMLIVERonCHIP project aims at developing the first in vitro model that reproduces both the liver and its associated vascular system.
Participating in this project are 4 partners with complementary expertise:
- The laboratory of Biomechanics and Bioengineering who have already developed and validated a liver on chip model. They will lead the development the endothelial cell chip and its connection to the liver chip.
- The Laboratory of Integrated Micro Mechatronic Systems who have strong expertise in endothelial cells, angiogenesis and vascular networks. They will grow primary endothelial cells and drive their growth into a relevant cell layer representative of the liver’s vascularisation.
- HCS Pharma is a CRO specialized in the development of innovative 2D and 3D cellular models for pharmacology and toxicology. They will validate the final liver-on-chip model by evaluating the effect of known substances and benchmark it with existing techniques.
- Fluigent will develop an automated instrument that will meet pharma standards in terms of throughput, automation, connection, compactness and ease of use.
The kick-off meeting took place on the 3rd of October. This project is very innovative and we expect to find significant results on the co-culture of the hepatocytes and LSEC (Liver Specific Endothelial Cells) on our way to the final report.
MYOCHIP FETOPEN PROJECT
The aim of the MyoChip project is to build a 3D human skeletal muscle irrigated by vasculature and innervated by neurons. The reconstituted 3D muscle will mirror the architecture and function found in vivo, namely in shape, contractility and microenvironment, while irrigation by a vascular network and innervation by human motor neurons will bring additional physiologic pertinence to it. This organ-on-a-chip technology will have numerous applications including but not limited to research on muscle building and aging, drug testing and screening, as well as prosthetics and biorobotics. The feasibility of the project relies on the interdisciplinary approach which joins a team of cell biologists, material engineers, experts in microfluidics and mathematical modellers. The architecture of skeletal muscle and its regenerative capabilities make muscle a prime candidate to push the 3D tissue engineering field. As such the project will lay the technical, material and methodological foundations to tackle the next generation of complex organ-on-a-chip systems that the MyoChip consortium can exploit for the generation of highly complex 3D in vitro systems of many organs.
BIO-ART LUNG 2020
The BIOART-LUNG 2020 project aims to develop a long term, autonomous, portable, artificial lung for patients suffering from acute respiratory distress. This innovative therapeutic approach will provide 2 major advantages compared to existing extracorporeal membrane oxygenation:
- Portable device that will increase mobility of patients while waiting for lung transplantation surgery
- Physiologically relevant device which would reduce blood activation and extended blood oxygenation above the current three weeks limitation.
To validate intermediate steps, Fluigent has conceived, developed and made a customized fluidic platform connected to the device that integrates all the functions the final system will have. This fully automated platform allows for testing of the devices under realistic conditions.
HOLIFAB
Microfluidics is currently faced with 2 main difficulties:
- A considerable hindrance to fast prototyping and industrialisation, because system performances rely on complex selection and assembly of numerous fluidic, mechanic, optical, electronic (etc…) components,
- A complex process for setting the system requirements, because constraints and specifications of microfluidic systems cover a huge range in terms of complexity level, materials, dimensions, acceptable costs, without any standardisation and rationalisation of design and production.
The HoliFAB project aims at overcoming these difficulties with a holistic system-oriented and problem-solving approach, starting from the customer’s need, optimising the chip and addressing the question of its environment.
HoliFAB change of paradigm implies:
- A holistic approach at the instrument level: Providing new methods and production tools to instruments manufacturers and new generation of applications to users,
- A holistic approach for the chip production: Using a technology-agnostic strategy integrating both 3D printing and injection moulding technologies for the design of the microfluidic chip.
INDEX
The aim of Project INDEX is to isolate and characterize nanoparticles available in bodily fluids through development and integration of novel technological breakthroughs. The technology will enable the analysis of clinically valuable nanoparticles called exosomes towards new generation diagnostics. Exosomes are known to mediate communication between cells and their effective utilization holds a great promise of revolutionizing the standard of clinical care. However, their detection and molecular profiling is technically challenging. The proposed technology will isolate exosomes that are as small as 30nm in diameter from human plasma with high purity, and provide in-depth, multi-parameter characterization of the particles through digital counting, size determination, and biological phenotyping.
Towards this goal: (1) Novel microfluidics will be developed and used for efficient magnetic enrichment; (2) Isolated particles will be detected and analyzed with a novel biological nanoparticle (BNP) sensor (3) Immune-capture and release chemistries as well as phenotyping assays will be developed; (4) Critically, complete on-chip integration of isolation, detection and analysis will be accomplished; (5) Utility of in-depth exosome characterization will be demonstrated with clinical samples for lung cancer.
Project INDEX requires successful integration of multiple sub-units and assays that each represents technological frontiers, which is extremely challenging. However, the breadth of information on exosomes that will be available with the integrated system is unmatched. Although, the clinical utility of exosomes is still developing, the uncertainty can only be clarified through automated technologies that provide latitude of information. Once completed, Project INDEX can demonstrate a new paradigm in cancer diagnostics, and also present a potential future technology for other applications involving nanoparticles.
MFMANUFACTURING
The objective of the MFManufacturing project is to bring the manufacturing of microfluidic devices to the same level of maturity and industrialisation of electronic devices, enabling them to address more widely in the healthcare needs. Electronic devices, which have been on the market for many years, have benefited from the long going standardization of electronic components, and were therefore easily integrated in the production process of the major foundries.
The anticipated standardization in the microfluidics field – first of all aimed at strengthening Europe’s position will focus on increasing maturity of both functional and fabrication process aspects:
- Gain of maturity in MF functions focusing both on novel functional modules and their interoperability.
- Gain of maturity in manufacturing process: focusing on a distributed pilot line, on novel hybrid integration processes and on increasing maturity of some selected manufacturing processes, which have a good short term commercial perspective.
This will have an effect on availability, reliability as well as accessibility and will result in device cost reduction and improved time-to market. These conditions will enable large scale uptake of microfluidic devices in the markets identified.
What is microfluidics?
Microfluidics is a field at the intersection of physics, engineering, and biology that has evolved over several decades. Microfluidics is both the science that studies the behavior of fluids flowing through micro-channels, and the technology of systems that process or manipulate small (10-6 to 10-12 L) amounts of fluids using microminiaturized devices containing chambers and channels through which fluids flow or are confined.
This discipline has been growing exponentially since the 1990s, and is viewed as an essential tool for life science research or biotechnologies more generally. It is a very attractive technology for both academic researchers and industrial groups because it considerably decreases sample and reagent consumption, shortens experiment times, and reduces the overall costs of applications.
Working principle of microfluidics
Microfluidics deals with very precise fluid control usings small volumes and spaces, with the “micro” prefix referring to one or more of the following features:
- Small volumes (µL, nL, pL, fL)
- Small size (mm, µm)
Microfluidic chips are the devices used in microfluidic studies in which microchannels have been molded or patterned. The microchannels are connected to allow fluids to pass through different channels, moving from one location to another.
Active microfluidics refers to fluid handling performed by active components such as microfluidic pumps or microfluidic valves. Microfluidic pumps, such as pressure-driven controllers, peristaltic or syringe pumps, supply fluids in a continuous way or are used for dosing, whereas microfluidic valves can inject precise volumes of sample or buffer.
Components of a microfluidic setup
A microfluidic system typically consists of various components designed to manipulate and control the flow of fluids on a microscale. Here are some common components found in microfluidic systems:
- Microchannels: Small, intricate pathways on a chip through which fluids flow. These channels are often fabricated using microfabrication techniques like photolithography.
- Reservoirs: The points where fluids are loaded into or collected from the microfluidic system. They act as source and sink locations for the liquids being manipulated.
- Microfluidic valves: They regulate the flow of fluids within microfluidic channels. They can be passive (relying on the geometry of the channels) or active (electronically controlled). Valves are crucial for directing and stopping fluid flow as needed.
- Microfluidic pumps: They generate pressure or flow to drive the movement of fluids through the microchannels. Various types of pumps, including syringe pumps or peristaltic pumps, may be integrated into microfluidic systems.
- Sensors: Optical or electrochemical sensors are integrated to detect and analyze the properties of the fluids, allowing for real-time monitoring and feedback.
- Detectors: They identify specific signals or changes in the fluids. Common detectors include photodetectors for optical analyses or electrodes for electrochemical sensing.
- Microfluidic chip: The microfluidic chip itself is a key component, acting as the physical platform on which all the channels, valves, and other elements are integrated. It is often made from materials like glass or polymers.
- Software and control system: Used to program and monitor the operation of the microfluidic system. It controls the various components, such as pumps and valves, to execute specific fluidic processes.
Some systems have additional components such as mixers, microvalves, detectors, temperature controllers, etc. These components work in harmony to create a microfluidic system capable of performing a wide range of tasks, from chemical analyses and synthesis to biological studies and diagnostics.
WEBINAR REPLAY – Overview of Microfluidics
This webinar aims to provide a broad introduction to microfluidics and related physical principles. The webinar will also feature some typical applications in microfluidics.
What you will learn:
- Introduction to microfluidics: Definitions and terminology, history and areas of application.
- Overview of common microfluidic systems:
- Microfluidic chips
- Microfluidic flow controllers
- Physical principles related to microfluidics:
- Laminar flow
- Diffusion in microfluidics and how to control mixing
- Droplet microfluidics:
- Definitions and related physics
- Droplet chip designs and droplet generation regimes
- Surfactants in droplet microfluidics
Webinar Replay – LineUp Series, the new generation of microfluidic controllers
Discover the LineUp™ Series, Fluigent’s leading fluidic delivery platform.
A complete solution, including software for automation, to easily adapt to any microfluidic setup and provide unmatched accuracy, simplicity, and versatility.
What will be covered?
Introduction to microfluidic systems and pressure-based flow controller.
Discover Fluigent’s revolutionary pressure-based systems
A complete overview of the LineUp™ series features and capabilities
Why you should attend:
Challenge your current microfluidic setup with Fluigent revolutionary solutions
Discover unsuspected features and modules of the LineUp series and its future developments
You are developing a new microfluidic experiment and wonder how Fluigent solutions can help you
Advantages and Key Principles of Microfluidics
Microfluidics is a very attractive technology offering an array of advantages for both academic researchers and industrial groups
At a micrometric scale, the behavior of fluids changes and presents several advantages: rapid heat transfer, an increased surface-to-volume ratio, laminar flow, and possible diffusive mixing. In addition, microfluidics considerably decreases sample and reagent consumption, shortens experiment times, and reduces the overall costs of applications.
The overall result is a significant increase in efficiency, driving a substantial reduction in both resource usage and overall application costs.
Microfluidics for Miniaturized Laboratories
A key concept related to microfluidics is the ability to integrate operations that would typically require a whole laboratory into a simple micro-sized system. Currently, a traditional scale-up is substituted in microfluidic systems by multiplexing, drawing on the compact size of the device to dramatically shorten the time from formulation to production. This leads to the adoption of microfluidic technologies not only for analytical purposes but also for large-scale manufacturing in process industries, particularly nanomedicine, fine chemistry, and the food, environmental, and pharmaceutical industries. [1]
Through the dynamic field of microfluidics, a new era of possibilities unfolds.
- Experimental accuracy: Microfluidics empowers researchers to refine and elevate the precision of scientific challenges, pushing the boundaries of detection to new lows and unraveling insights at the molecular level that were once unimaginable.
- High efficiency: Microfluidics allows for parallelized analyses, unlocking the ability to run multiple experiments simultaneously to achieve remarkable results with unparalleled efficiency.
- Cost reduction: Microfluidics offers a gateway to cost reduction without compromising the quality of the work.
- Time reduction: Often the most precious commodity in research, time becomes a valuable ally with microfluidics and its potential to accelerate the pace of discovery by significantly shortening experimental timelines.
- Precision meets practicality: Microfluidics is reshaping the landscape of research and industry where efficiency becomes the hallmark of progress. It has limitless possibilities and applications in various fields.
Precise control and automation
Microfluidic systems also offer excellent data quality and improved parameter control, allowing for process automation while maintaining high performance. They have the ability to both process and analyze samples with only minor sample handling. The microfluidic chip is combined with a fluid handling system to achieve incorporated automation that allows users to generate multi-step reactions requiring a low level of expertise and a wide range of functionalities.
Origins of microfluidics, a fluid handling system for printers
The history of microfluidics dates to the 1950s, principally in inkjet printer manufacturing. The mechanism behind these printers is based on microfluidics, and involves the use of very small tubes carrying the ink for printing.
In the 1970s, a miniaturized gas chromatograph was constructed on a silicon wafer. By the end of the 1980s, the first microvalves and micropumps based on silicon micro-machining had also been presented. In the following years, several silicon-based analysis systems were presented.
All of these examples represent microfluidic systems, since they enable the precise control of decreasing fluid volumes on one hand, and involve miniaturization of fluid handling systems on the other.
A major contribution in this field has been the development of soft lithography in a fast prototyping polymer, polydimethylsiloxane (PDMS), as a method for fabricating prototype devices and testing new ideas.
The expansion of microfluidics and development of microfluidic components
In the 1990s, advancements in microfabrication technologies expanded the possibilities of microfluidic systems. Researchers began designing lab-on-a-chip devices for applications ranging from chemical analysis to medical diagnostics. This era saw the emergence of the first microfluidic devices with integrated sensors and valves.
As the 21st century unfolded, microfluidics experienced a surge in popularity. The technology found applications in genomics, proteomics, drug discovery, and point-of-care diagnostics. Researchers explored the potential of organ-on-chip models, replicating human physiological conditions for more accurate testing.
Today, microfluidics continues to push boundaries, with ongoing research focusing on enhancing precision, scalability, and integration with other scientific disciplines.
Over the years, researchers spent a lot of time developing new microfluidic components for fluid transport, fluid metering, fluid mixing, valving, or concentration and separation of molecules within miniaturized quantities of fluids.
In 2006, for example, Fluigent was the first company to introduce a disruptive new way of handling fluids in microfluidics: microfluidic pressure pumps.
The use of a pressure-based pump instead of a syringe pump allows for very rapid response times and pulseless flow. At first, these pumps could only control the pressure of liquids in microfluidic chips, but later, by adding a flow sensor and a unique feedback control loop, Fluigent enabled control of both pressure and flow rate. The precise control of fluids in a microfluidic device allows for sophisticated new applications which were not possible before.
Recently, an increasing number of microfluidic-based devices, developed by both small start-ups and large pharmaceutical and biomedical companies, have been released and are entering the market.
Comparative example: Why choose a microfluidic device instead of a robot?
Due to the low volumes required, microfluidic technologies represent a promising alternative to conventional laboratory techniques. They allow for complete laboratory protocols on a single chip of a few square centimeters. Table 1 shows the main advantages of using microfluidics instead of conventional laboratory assays for a given experiment (ultra-high throughput screening of a typical enzyme).
Robot | Microfluidic droplets | |
---|---|---|
Total reactions | 5 × 107 | 5 × 107 |
Reaction volume | 100 µL | 6 pL |
Total volume | 5,000 L | 150 µL |
Reactions/day | 73,000 | 1 × 108 |
Total time | 2 years | 7 hours |
Number of plates/device | 260,000 | 2 |
Cost of plates/device | $ 520,000 | $1.00 |
Cost of tips | $10 millions | $0.30 |
Amortized cost of instruments | $ 280,000 | $1.70 |
Substrate | $4.75 million | $0.25 |
Total cost | $15.81 million | $2.50 |
Table: Comparison using traditional methods and in microfluidic emulsions. Adapted with permission from Agresti J. J. et al, Ultrahigh-throughput screening in drop-based microfluidics for directed evolution, PNAS 2010, 107:4004-4009. Copyright 2010 National Academy of Sciences, U.S.A [3]
To better appreciate the impact of microfluidics, we can draw an analogy with the evolution of computers. In the 1960s, an entire room was needed to run a computer. Since then, every component has been reduced in size, and laptop products have appeared. Now a simple smartphone is more powerful than any computer built before, leading to reduced prices and a much more user-friendly experience. It’s the same with microfluidics!
Overview of microfluidics applications
Microfluidics is an attractive technology for multiple fields.
Microfluidics extends its capabilities in precise liquid injection to the fields of cell perfusion applications and droplet generation, breaking free from the confines of traditional “lab on a chip” and “organ on a chip” technologies.
Microfluidics, renowned for its intricate control over small fluid volumes, overcomes conventional boundaries and finds applications across a range of fields.
- In cosmetics, microfluidics takes center stage, enabling precise crafting of emulsions and formulations and revolutionizing product development through increased efficiency and accuracy.
- In pharmaceuticals, especially in drug discovery, microfluidics accelerates experimentation, providing a sophisticated and resource-efficient approach.
- The healthcare sector benefits significantly from microfluidics, where the technology contributes to personalized medicine and diagnosis. Its precision in manipulating small fluid quantities opens the door to novel diagnostic approaches and tailored medical interventions.
- In chemistry, microfluidics has emerged as a tool for flow synthesis and stoichiometry, optimizing chemical processes.
- Biologists leverage microfluidics for cell culture and 3D printing, creating environments that closely mimic physiological conditions.
- Droplet generation applications further extend its versatility, with microfluidics emerging as a pivotal tool for creating and manipulating droplets with high precision.
- In energy applications, microfluidics plays a crucial role in Enhanced Oil Recovery (EOR) models and plasma confinement studies, showcasing its adaptability across different scientific disciplines.
- In industrial applications, microfluidics provides a platform for enhanced efficiency, precision, and cost-effectiveness.
Draw on microfluidic technology for your industrial application
- High-Throughput Screening: Microfluidic devices excel in conducting rapid and parallelized experiments, making them ideal for high-throughput screening in industries like pharmaceuticals and biotechnology. This enables quick testing of multiple conditions, reducing time and resource requirements.
- Process Miniaturization: Microfluidic systems allow for miniaturization of processes, leading to reduced sample and reagent consumption. This not only cuts costs, but also facilitates handling of scarce or expensive materials.
- Point-of-Care Diagnostics: Microfluidics plays a crucial role in developing portable and rapid diagnostic tools for on-site testing. These devices can be utilized in industrial settings for real-time monitoring, ensuring quality control and minimizing downtime.
- Customized Manufacturing: Microfluidics allows for the creation of tailored microenvironments to facilitate customized manufacturing processes. This is particularly beneficial in applications where specific conditions are required for optimal product development.
Custom box: Engineering solutions
- Automation and Integration: Microfluidic components can be easily integrated into automated systems, streamlining processes and reducing the need for manual intervention. This enhances overall workflow efficiency in industrial applications.
By leveraging the capabilities of microfluidics in these ways, industries can enhance their processes, reduce costs, and improve overall productivity.
The development of microfluidics has just begun!
Conclusion
Microfluidics offers revolutionary new capabilities. It is still quite a new technology, and there is a lot of work to be done so that it can solve problems for users who are not experts in fluid physics, such as clinicians, cell biologists, and public health officials.
Microfluidic applications and products are already present in the marketplace, especially in nanomedicine, allowing for more precise analysis of molecules like DNA and proteins, bacteria, or analysis at the scale of a single cell. The continuing development of high-throughput screening and organ-on-chip technology will lead to faster and better drug development. With the development of lab-on-a-chip and microTAS and the possibility of combining microfluidics with automation, new diagnostic products will be cheaper and faster, bringing benefits to developing countries.
Expertises & Resources
References:
- Fluigent, White paper: An exploration of microfluidics and fluid handling, 2020, https://www.fluigent.com/resources-support/expertise/white-papers/
- Bahnemann, J.; Grünberge, A. Microfluidics in Biotechnology: Overview and Status Quo. Advances in Biochemical Engineering/Biotechnology book series, 2022, ABE,volume 179.
- Agresti, J. J.; Antipov, E.; Abate, A. R.; Ahn, K.; Rowat, A. C.; Baret, J.-C.; Marquez, M.; Klibanov, A. M.; Griffiths, A. D.; Weitz, D. A. Ultrahigh-Throughput Screening in Drop-Based Microfluidics for Directed Evolution. Proc. Natl. Acad. Sci. U.S.A. 2010, 107 (9), 4004–4009.
About The Keyser lab – University of Cambridge
The Keyser Lab is a group of researchers at the Cavendish Laboratory, University of Cambridge, UK. Since its founding in 1874, the Cavendish Laboratory has been at the forefront of discovery in physics, with a core focus on experimental physics supported by excellence in theory. The department promotes world-leading experimental and theoretical physics in all its diversity. Scientists from the Keyser Lab study the physics of ions, macromolecules and particles, with a particular focus on particles in confined geometries at the single molecule/particle level. To exert maximum control over all parameters for their experiments, they make use of several cutting-edge techniques such as DNA self-assembly (origami), optical trapping, electrophysiology, and microfluidics and nanofluidics.
The team includes researchers with expertise in physics, engineering, physical chemistry, biochemistry/biology, and micro- and nanofabrication.
What are Giant Unilamellar Vesicles (GUVs)?
Giant Unilamellar Vesicles (GUVs) are micron-sized compartments composed of lipid bilayers, serving as models for cell membranes or as encapsulating structures for biological materials within cell-like environments. These vesicles have dimensions ranging from 1 to 100 µm, mimicking the size of cells. Like natural cell membranes, GUVs are constructed from lipids, predominantly phospholipids and cholesterol. The amphiphilic properties of these lipids enable them to spontaneously arrange into spherical compartments when immersed in an aqueous solution.
They offer the advantages of having well-defined lipid compositions, being easy to image, and being controlled systems for studying transport processes. These characteristics open the door to a variety of applications in biology and biomedicine, especially membrane biophysics and synthetic biology.1,2
Working principle of GUV production
In 2016, the Dekker Laboratory from the Delft University of Technology developed a novel microfluidics-based method called Octanol-Assisted Liposome Assembly (OLA) to create uniform, cell-sized (5–20 mm) GUVs with high encapsulation efficiency. Using Fluigent’s pressure-driven MFCS-EZ controller with the Oxygen software tool, an inner aqueous phase (IA) and an outer lipid-carrying 1-octanol phase (LO) were combined, resulting in double-emulsion droplets via hydrodynamic flow focusing. These droplets developed a side-connected 1-octanol pocket, which, due to interfacial energy minimization, separated to rapidly form fully assembled solvent-free liposomes. Microfluidic GUV production addresses the persistent issue of residual oil in vesicle bilayers.
This method allows researchers to generate GUVs with higher monodispersity and greater size control compared to traditional methods, and with a faster process than alternative microfluidic methods.3
Keyser Lab: Microfluidics for GUV generation for antimicrobial efficacy and biomimetic vesicle membrane testing
Scientists from the University of Cambridge integrated octanol-assisted liposome assembly into their microfluidic platforms (“lab on a chip” devices) for quantifying drug permeation and antimicrobial efficacy on biomimetic vesicle membranes.
In a first paper published in Lab on a Chip (2019), they reported on a microfluidic platform for testing antimicrobial peptides on artificial vesicle membranes. This platform produced vesicles with an encapsulated dye to assess the efficacy of antimicrobial peptides by measuring the time it takes for vesicles to lyse.4
This Giant Unilamellar Vesicle production technique also utilized Fluigent‘s MFCS-EZ and its accompanying software (Oxygen) for fluid control. Vesicle formation and perfusion inlets were connected, and the microfluidic chip was loaded with an inner aqueous phase base stock. The outlet was connected to Fluigent’s 2-Switch, which can switch between open or closed configurations for waste removal. Vesicles flowed through a channel to the connector chip, and 1-octanol pockets pinched off to form droplets. Density-based separation removed vesicle production waste, and the vesicles were distributed to trapping chambers. Peptide doses were controlled, replacing the inner aqueous phase buffer, while maintaining a constant input pressure. The entire process was monitored under an inverted microscope.
Validated with cecropin B on bacterial-mimicking membranes, the platform enabled the researchers to study over 1000 vesicles simultaneously. The results showed dose-dependent disruption of vesicle membranes, demonstrating the platform’s potential for controlled, quantitative assessments of the efficacy and selectivity of antimicrobial peptides. This microfluidic approach to GUV production is suggested as a new standard for pre-clinical development of membrane-active antimicrobials, offering advantages in cost efficiency and parallelization.
In another paper published in Biomembranes (2020), scientists from the University of Cambridge and the University of Exeter produced GUVs with tunable binary lipid mixtures to determine lipid diffusion in OLA vesicles.5
The focus was on expanding the capabilities of this microfluidic approach to form GUVs with tunable binary lipid mixtures. An MFCS-EZ equipped with a Fluiwell-4C reservoir kit with OLA solutions was connected to the microfluidic chip. The microfluidic GUV production method allowed for a high degree of control over vesicle sizes by adjusting pressures in different channels. This was possible due to the high responsiveness, stability and repeatability of the pressure generated by the MFCS-EZ. In addition, flow speed was successfully matched to the outlet channel length to prevent residual octanol attachment to vesicles.
This study employed fluorescence recovery after photobleaching to investigate lipid lateral diffusion coefficients in GUVs produced by the microfluidic approach, finding values within the expected range. Comparisons with electroformed vesicles indicated quantitative similarity in lipid diffusion coefficients. The results served as a quantitative biophysical validation of OLA-derived GUVs, enhancing the potential application of this versatile platform in drug discovery, artificial cell production, and lipid membrane studies.
Conclusion
To produce Giant Unilamellar Vesicles, fluids were controlled using Fluigent pressure-based flow controllers. Researchers from the Keyser Lab (University of Cambridge) typically operated the chip with input pressures of 40 mbar for the inner aqueous phase and dissolved-lipid phases and 100 mbar for the outer aqueous phase. For precise flow measurements, Fluigent Flow Units can be added on the fluidic path. Microfluidic GUV production also allowed researchers to adjust the sizes of the generated vesicles by adjusting the microfluidic pressures of the phases. This degree of control is difficult to achieve using standard methods such as electroformation or traditional syringe pumps.
Testimonials
“Microfluidics presents various advantages to researchers who need small volumes and high throughput in answering their scientific questions. In our lab, we use microfluidic devices for standardization and control of experimental parameters like concentration and timing. In the complex (biological) systems we are working on, the mentioned characteristics are fundamental in collecting reliable meaningful statistics, and microfluidics in combination with light microscopy offers just that. We also heavily rely on the ability to rapidly prototype devices, as we can design bespoke solutions at minimal production cost and time.”
“We use the pressure-based pumps from Fluigent for experiments that require swift responsiveness when manipulating fluids, and fine tuning at low flow rates. We use the Fluigent systems during fabrication and running of the microfluidic chips. The ability to pump in air at high precision makes the Fluigent pressure-based systems ideally suited to selectively coat and functionalize micro-channels within a microfluidic network. After coating we then fill the devices with the experimental solutions and use the pressure controls to move fluids around, open and close valves and carefully time the introduction of small molecules in the experiments.“
Kareem Al Nahas, University of Cambridge
Expertises & Resources
References
- Naziris, N., Demetzos, C. (2021). Liposomes: Production Methods and Application in Alzheimer’s Disease. In: Vlamos, P. (eds) GeNeDis 2020. Advances in Experimental Medicine and Biology, vol 1339. Springer, Cham.
- Pereira, David & Valentão, Patrícia & Andrade, Paula. (2014). Nano- and Microdelivery Systems for Marine Bioactive Lipids. Marine Drugs. 12. 6014. 10.3390/md12126014.
- Deshpande, S.; Caspi, Y.; Meijering, A. E. C.; Dekker, C. Octanol-Assisted Liposome Assembly on Chip. Nat Commun 2016, 7 (1), 10447.
- Al Nahas, K.; Cama, J.; Schaich, M.; Hammond, K.; Deshpande, S.; Dekker, C.; Ryadnov, M. G.; Keyser, U. F. A Microfluidic Platform for the Characterisation of Membrane Active Antimicrobials. Lab Chip 2019, 19 (5), 837–844.
- Schaich, M.; Sobota, D.; Sleath, H.; Cama, J.; Keyser, U. F. Characterization of Lipid Composition and Diffusivity in OLA Generated Vesicles. Biochimica et Biophysica Acta (BBA) – Biomembranes 2020, 1862 (9), 183359.