Long-term perfusion studies
Automated Sequential Injection System[OAR]
Fluigent introduces the Aria, our automated sequential injection system for cellular perfusion or timed injection protocols. The Aria allows users to automate delivery of up to 10 different solutions into a chamber or microfluidic chip following user-defined protocols.
For cellular perfusion, long-term imaging of cultured cells requires controlled environmental conditions. Continuous fluid flow injection of the medium makes it possible to maintain cells under optimal physiological conditions, with constant nutrient supply and a controlled pH environment while avoiding cell debris accumulation.
Our multiple fluid injection system generates minimal shear stress for cells, allows for multiple fluid switching and enables users to achieve medium flow continuously through pressure variations. Additional components in our fluid injection system ensure a highly controlled flow rate through flow sensors, and the possibility to perform fast-medium switches.
- Save time
- Easy to use
All in one instrument and software
Fits any experimental design
Features of Aria
Deliver up to 10 solutions
The Aria can be used for any application, as the volumes injected range from 40µL to hundreds of mL delivered over several days. The tubing can be cleared to minimize carryover between successive solutions. The user is informed of the delivery time for each solution thanks to Aria’s intelligent software. The software also notifies the user of the minimum volume required in each reservoir to run their protocol.
Automate any protocol
The Sequential Injection System comes with intuitive software that lets users design protocols in just a few clicks. Incubation time, flow rate, and volume dispensed are all parameters that can be easily set by the operator for each step of the injection protocol. Protocols are recorded and can be shared among users. Flow rate and pressure are recorded for each experiment.
The Aria drastically reduces variability to as little as 0.5% between experiments, compared to 5.1% intra-operator variability and 8.1% inter-operator variability using a pipette.
Preserve sample integrity
Contamination free: the sample is delivered at the beginning of the experiment and is not touched during the protocol, minimizing contamination risks from manual operations. This results in controlled and smooth flow, preserving samples more effectively than with traditional pipetting.
Ideal for imaging studies
The Aria can synchronize with various microscopes using TTL signaling. The device can send and receive TTL signals to start an imaging cycle, or to resume the Aria injection protocol when an imaging cycle is completed. Aria’s software comes with a dark mode to work in an imaging room, and is equipped with LEDs to facilitate use in dark areas.
Radtke, AJ, et al. IBEX: an iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues. Nature Protocols (2022) doi: 10.1038/s41596-021-00644-9
Huang, K. et al. A Novel Method to Map Small RNAs with High Resolution. Bio-protocol (2021) doi: 10.21769/BioProtoc.4128.
Radtke, AJ, et al. IBEX: An open and extensible method for high content multiplex imaging of diverse tissues, Presented at CZI HCA network and HuBMAP consortium
Why use an instrument for automating sequential injections?
Applications like DNA hybridization, high resolution spatial transcriptomics, sequential imaging, DNA paint, labeling or tissue profiling require high resolution microscopy, as they target nanometric structures inside the cells.
For each of these applications, the sample is successively incubated with specific probes to bind to a target inside the cells. Between the different probes, a sequence of washing steps is performed to remove the previous probe and avoid contamination with the next one.
The power of this fluid/injection technique compared to traditional immunolabeling is that the number of targets per sample are not limited to the four traditional colors of the microscope, as DNA can be easily detached from its target, as opposed to antibodies that remain strongly attached when bonded.
WEBINAR: Enhancing Microfluidic Cell immunolabeling with Aria Technology
Discuss approaches to automating the cellular immunolabeling process using microfluidic devices, thereby increasing efficiency and reproducibility.
- Introduction to Fluigent’s expertise in the field of microfluidics and Organ-on-chip
- Aria: Fluigent’s automated sequential injection system
- Success story using Aria for neuron immunolabeling
- Have a live discussion with our experts and the option to discuss specific applications
WEBINAR REPLAY – Automating Cellular Studies with Aria
Watch the webinar by our team about Automating Cellular Studies with the Aria Automated sequential injection system.
Get the success you deserve with Fluigent’s new perfusion system, Aria, that takes care of fluid delivery for you.
If your experiments require precise timed delivery of reagents, probes, fluorophores, etc. while minimizing operator interaction, look no further than Aria, our next-generation instrument for automating cellular perfusion studies.
Based on input from our customers, protocols requiring delivery of more than one solution require large amounts of time to set up, as well as significant operator interaction.
The Aria system provides rapid experimental setup, ease of use, and can automate even multi-day protocols. It can interface with many microscopes to further simplify the process of reagent delivery and imaging. Aria is the best solution to automate your lab work. Highly flexible, it can adapt to any perfusion chamber and any protocol thanks to its intuitive software. Aria is also a tool for automating complex protocols for live-cell imaging (immunostaining, omics applications, radiometric imaging).
Webinar hosted by:
Marine Verhulsel: Product Manager, Fluigent
William César: R&D Project Manager, Fluigent
Mathieu Steiner: Software Engineer, Fluigent
WEBINAR REPLAY – Automated fluid delivery for cell and tissue imaging
Want to gain time, precision and reproducibility for your immunofluorescence assay, or any other assay requiring injection of multiple solutions on your sample? Watch our webinar to learn how to interface a flow chamber (or microfluidic chip) to our automated fluid delivery device ARIA. This will allow you to deliver up to 10 different solutions in a sequential and autonomous manner. In addition, ARIA can be synchronized with any microscope to launch an image acquisition cycle and resume the perfusion protocol once the imaging cycle has been completed.
This all-in-one workflow facilitates alternation between cycles of injection/incubation time with reagents and image acquisition, a feature particularly well-adapted to complex cell and tissue imaging. Here we will present some example applications such as multiplexed tissue imaging, DNA-PAINT and seqFISH, as well as cell capture and staining.
00:00 – Reasons for & advantages of using automated sequential fluid delivery
08:30 – Demo video
10:27 – Example applications:
- Automated multiplexed tissue imaging
- Sequential fluorescence in situ hybridization (SeqFISH)
- Capture and characterization of circulating tumor cells
26:40 – Q&A session
Webinar hosted by:
Mélanie Chabaud (Fluigent – Application Engineer)
William César (Fluigent – R&D project manager)
Noé Viovy (Fluigent – R&D software developer)
WEBINAR REPLAY – How to turn your fluorescence microscope into a spatial omics platform
Current approaches in genomics, transcriptomics, and proteomics yield quantitative abundance analysis of biomolecules on an almost routine basis, with a critical impact in the life sciences.
However, coupling this high content to spatial information in a single-cell and tissue context is still a challenge, and this is where our efforts are presently focusing.
In this webinar, I will share my facility’s experience in building spatial omics platforms.
First, I will provide an overview and comparison of microscopy-based methods for spatial omics. Then, I will present details and resources on how to build such a platform.
Finally, I will show the existing tools available for image and data analysis associated with these methods.
It is my hope that our experience can serve others in the road ahead to help decide which technique is best for their applications and to assist in their implementation.
Webinar hosted by:
Alvaro Crevenna: Head of Microscopy, EMBL Rome
Dale Clark: Chief Operating Officer, Bioptechs Inc.
Reduce experimental error with our Automated Sequential Injection System
Be aware of the following challenges when manually changing solutions from washing buffers to probes on the microscope.
- A touch of a pipette tip can displace the dish and recorded positions
- Samples can be flushed away during pipetting
- The sample can get contaminated
- Liquid can spill over the microscope
- Errors in the sequence of solutions due to the number of steps
In addition, without a Sequential Injection System, we will have to perform successive solution replacements that extend over long periods (up to 4-5 days for an experiment) and require the physical presence of a scientist.
|Type of injections||Abrupt injection (up to 1mL in few seconds)|
|Smooth & controlled injections|
|Geometry at injection tip||Conic shape: important shear strain|
Unhomogeneous fluid velocity
|Straight shape: No modification at the injection tip|
Automate any protocols
Aria is the only instrument that can automate the delivery of up to 10 different solutions to a flow cell. It’s the perfect compromise between manual pipetting and an all-in-one system dedicated to one specific application. Any protocol with multiple solution delivery can be automated, saving the scientist time and reducing variability between experiments compared to procedures that do not include perfusion systems. Users can integrate their own microscope, specific chip type and solution sets using the automated injection platform.
- Aria is straightforward and ready to use. No specific training is required, and no time is lost troubleshooting incidents or manually priming solutions before the experiment as with custom-built systems.
- Compared to fully integrated systems that are bound to one chip type and one set of solutions for a given application, Aria offers flexibility. Aria can automate different applications in the same lab, as it is compatible with any chip and any type of reagent. Aria is transportable and can be easily moved between microscopes.
- Aria is a cost-effective alternative to an all-in-one system. It is perfect for investigating and setting all the parameters to fully optimize perfusion culture for live-cell imaging.
Seamless integration through our dedicated SDK
In addition, an SDK library consisting of all available software functions is available for users that wish to integrate our liquid handling functions into their own software for OEM automation workflows. This allows users to have a single interface to actuate all components that are part of their fluorescence microscopy system (fluid management unit, imaging unit, heating devices, incubator, etc.).
“I got to test Aria automated sequential injection system in my research project in a collaboration with Fluigent. More precisely, Aria injection system helped me automatize the capture process and immunostaining of breast cancer cells under a very precise and controlled flow rate. The software interface is so user-friendly that I was being able to follow in real-time the progression of my experiment. The amazing part is that ARIA even calculated the total amount of time required for each step and helped me avoid the waiting time in front of my setup! It made my experiment go as smoothly as possible.”
Emile Lakis / Curie Institute, IPGG / Paris
Choose the version that best meets your needs
Single output version
Perfect for high quality multiplexed imaging experiments.
Aria is the perfect compromise between manual pipetting and all-in-one system that are dedicated to one specific application, integrate a microscope, specific chip type and a given set of solutions: Any protocol with multiple solution delivery can be automated, saving the scientist both time and reducing variability between experiments compared to manual procedures.
Serial output version (perfuse up to 9 samples)
Perfect benchtop workstation to automate staining or any routine protocols with multiple fluid delivery.
Aria is designed to fully automate your protocols, gain productivity, reliability, reduce manual operation and save time. Aria is the only workstation on the market compatible with any perfusion chamber.
Perfect for high-quality multiplexed imaging experiments
Aria is the perfect compromise between manual pipetting and all-in-one systems that are dedicated to one specific application, integrating a microscope, specific chip type and a given set of solutions. Any protocol with multiple solution delivery can be automated, saving the scientist time and reducing variability between experiments compared to manual procedures.
“Just wanted to say thanks again. We were able to run a 50 step Aria protocol on four separate occasions this weekend. Saved us more than a full day of work (~28 hours).”
M. Serrata / Wyss Institute / Boston, Massachusetts
|Flow rate control||Over the range of 40 µL/min to 1 mL/min for water|
|Pressure control of flow rate||to a maximum of 2 bar|
|Valves||ten position switching valve – two position switching valve (2)|
|Fluid reservoirs (8)||15 mL standard, 2 mL available|
|Flushing solution reservoir (2)||100 mL|
|Tubing||FEP with OD of 1/16 inch and ID of 250 µm|
|Wetted surfaces||Polypropylene, FEP, Glass, PEEK|
|Compressed air source||Requires non-corrosive, non-explosive, biocompatible compressed air (lab line, gas|
cylinder, compressor or Fluigent FLPG)
|PC specifications||Windows 7 or higher|
|Dimensions||382 mm x 240 mm x 265 mm|
|Power supply voltage||24V DC|
|Gas compatibility||Dry, oil-free gas, air, any non corrosive or non explosive gas|
|Liquid compatibility||Aqueous solutions only|
|Control in real-time, protocol automation, data record and export|
|The SDK library collects the available software functions for users that wish to integrate our liquid handling functions into their own software for automated workflows, allowing them to have a single interface to actuate all components that are part of their fluorescence microscopy system (fluid management unit, imaging unit, heating devices, incubator, etc.).|
To save time and minimize reagents consumption, Aria loads its internal and external tubing with the successive solutions to inject in the chip.
If the protocol commands to inject 40 µL of solution A, incubate for 1 hour, inject 50 µL of solution B, incubate for 1 hour then wash with buffer for 5 minutes and the total volume of tubing ( internal + external) is of 300 µL Aria will prefill tubing as follows:
As a consequence, calibration of the tubing length and related volume is necessary.
Here is a description of all the functions available to optimize protocol writing in Aria
Calibration is automated and the software assists the user to determine the total volume of his step up (internal volume + L1+L2). Calibration values are recorded for future experiments.
Initially all tubing inside Aria are dry. The “Prefill” function is recommended as a first step to load all solutions inside Aria without injecting air in the chip or chamber. This function can be deactivated if the user performed the loading manually. Fluigent strongly recommends using this function.
Perfusion can be set in terms of volume (ex: step 1) or duration (ex: step 2) of injection.
The user selects the reservoir of the solution to inject (ex: reservoir 1), sets the flow rate (ex: 100 µL/min) at which the solution will enter the flow cell and the volume (ex: 100 µL) or duration (ex: 1min) of injection. The flow rate range is from 40 µL/min to 1 mL/min. The software automatically calculates the time at which the solution should enter the flow cell.
Incubation time can be easily set by entering the duration of the incubation (ex: 1h30) in the ‘wait’ function.
The ‘wait for user’ function is a variation of the ‘wait’ function. It is particularly useful if the user has to perform a manual operation like preparing cells before injection. The system waits until the user notifies it can proceed to the next step. In absence of notification by the user, the system proceeds to the next steps after 12 h.
To prevent contamination between successive injections, the tubing can be cleared with buffer using ‘Flush tubing’ function. The user selects the reservoir containing the wash solution (ex: reservoir 10) and the flow rate at which the tubing is flushed (ex: 100 µL/min).
Aria pushes all the residual fluids contained in the L1 tubing to waste, and fills the L1 with wash solution. Buffer contained in L1 will also be directed to the waste. This operation does not involve flow to the chip as neither residual fluids nor buffer enter the chip. However, the residual fluid contained in L2 will not be cleared. For this reason, Fluigent recommend the user to keep L2 as short as possible.
Aria is equipped with TTL and TCP signalling and can send and receive both signals.
- Sending TTL or TCP
Each step is flanked by two bells () : at the front and back of the settings (see below for ‘volume injection step).
By clicking on each of both of the bells they get activated (). Aria will then send a TTL or TCP signal either when the step starts (example below) or ends or both.
Fluigent recommends using this function to synchronize perfusion and imaging.
- Receiving TTL or TCP
The ‘Wait for Signal’ function puts the Aria on hold. The Aria then waits for an external signal, either TTL on input port or TCP, to run the next steps. If the Aria does not receive signaling after 12 hours, it automatically proceeds with next steps.
Minimum volume reservoir
Aria is designed to minimize reagent consumption. A small additional volume of liquid (36µL) is necessary to preload the system and to ensure that some residual liquid is still present in reservoir after injection to prevent delivery of air to the system.
As the user writes his protocol, the software calculates the minimum volume that should be placed in each reservoir to successfully run the protocol.
Write, load, save sequence
To design a new sequence, click on .
Before running a sequence, the software will automatically ask the user to save it. All saved sequences are accessible and can be loaded by clicking on .
Sequences can be saved at any time by the user by clicking on the saved data .
For each experiment, Aria automatically records and saves the flow rate, pressure, the reservoir from which the solution is withdrawn if solution is delivered to the chip or waste. All data is accessible by clicking on .
Expertise & resources
Fluigent products manual Aria SDK User Manual Download
Microfluidics case studies University of Rochester: A tissue chip platform for real-time sensing of secreted inflammatory markers using ARIA Read more
Fluigent Products Datasheets Aria Technical Specifications Download
Tutorial videos ARIA Tutorial Episode 1 : How To Plug An ARIA [automated cell perfusion] – Fluigent Read more
Tutorial videos ARIA Tutorial Episode 2 : Connect To Set The Fluidic Path [automated cell perfusion] – Fluigent Read more
Tutorial videos ARIA Tutorial Episode 3 : How To Fill Up The Reservoirs [automated cell perfusion] – Fluigent Read more
Tutorial videos ARIA Tutorial Episode 4 : How To Use ARIA Manually [automated cell perfusion] – Fluigent Read more
Tutorial videos ARIA Tutorial Episode 6 : Global Handling Of ARIA [automated cell perfusion] – Fluigent Read more
Tutorial videos ARIA Tutorial Episode 5 : Perform A Calibration With ARIA [automated cell perfusion] – Fluigent Read more
Tutorial videos ARIA Tutorial Episode 7 : Software Presentation [automated cell perfusion] – Fluigent Read more
Fluigent products manual Aria User Manual Download
Microfluidic Application Notes Cancer Cell Analysis Made Easy with Aria: cell Capture and Labeling Read more
Microfluidic Webinars Automate Cellular Studies with Aria Read more
Fluigent Products Datasheets Aria datasheet Download
Expert Reviews: Basics of Microfluidics Passive and active mechanical stimulation in microfluidic systems Read more
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