Long-term perfusion studies
Aria, Automated Sequential Injection System[OAR]
Automated system for sequential injection experiments
Fluigent introduces the Aria, our Automated sequential injection system for cellular perfusion or timed injection protocols. The Aria allows one to automate delivery of up to 10 different solutions into a chamber or microfluidic chip by following user defined protocols.
For cellular perfusion, long-term imaging of cultured cells requires controlled environmental conditions. Continuous perfusion of the medium allows maintaining cells under optimal physiological conditions, with constant nutrient supply and controlled pH environment while avoiding cell debris accumulation.
Our perfusion system generates minimal shear stress for cells and allows users to achieve medium flow continuously through pressure variations. Additional components in our perfusion 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
What are the features of Aria’s Automated sequential injection system?
Deliver up to 10 solutions
The Aria covers any applications as the volumes injected range from 40µL to hundreds of mL deliver over several days. To minimize carryover between successive solutions, the tubing can be cleared. The user is informed of the delivery time of 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 Aria Automated sequential injection system comes with intuitive software that enables to design protocols in 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 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 down to 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 preserve samples compared to traditional pipetting.
Ideal for imaging studies
The Aria can synchronize with various microscopes using TTL signalling. 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 LED’S to facilitate use in dark areas.
Why Aria, Automated sequential injection system?
Applications like DNA hybridization, high resolution spatial transcriptomics, DNA paint, labelling 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 various probes, a sequence of washing steps is performed to remove the previous probe and avoid contamination with the consecutive one.
The power of this perfusion/injection technique compared to traditional immunolabeling is that the number of targets per sample are not limited to the four traditional colours of the microscope as DNA can be easily detached from their target as opposed to antibody that remain strongly attached when bounded.
Watch the webinar by our team about Automate Cellular Studies with the Aria Automated sequential injection.
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 an Automated sequential injection system, we will have to perform successive solutions replacements that can be long (up to 4-5 days for an experiment) and require the physical presence of a scientist.
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 both 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 Aria Automated sequential injection system.
- Aria is straightforward and ready to use. No specific training is required, and no time is lost trouble shooting incidents or manually priming solutions before the experiment as is the case with home-made systems.
- Compared to fully integrated systems that are bound to one chip type and a set of solutions for a given application, Aria offers flexibility. Aria can automate different applications in one lab as it’s compatible with any chip and any type of reagent. Aria is transportable and can be easily moved between microscopes.
- Aria Automated sequential injection system is a cost-effective alternative to an all-in-one system. It is perfect to investigate and set all the parameters to fully optimize perfusion culture for live-cell imaging.
“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.
“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|
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 signalling and can send and receive external TTL signals.
- Sending TTL
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 signal either when the step starts (example below) or ends or both.
Fluigent recommends using this function to synchronize perfusion and imaging.
- Receiving TTL
The function ‘wait for TTL’ puts Aria on hold until it gets an external TTL signalling to proceed with the next steps of the protocol. If the Aria does not get any signalling it automatically proceeds with the next steps after 12h.
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
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
User manuals Aria User Manual Download
Microfluidic Application Notes Capture and Labeling of Cancer Cells Using Aria Read more
Microfluidic Webinars Automate Cellular Studies with Aria Read more
Microfluidic Webinars How to turn your fluorescence microscope into a spatial omics platform Read more
Technical datasheets Aria datasheet Download
Expert Reviews: Basics of Microfluidics Passive mechanical stimulation induced by laminar and pulsatile shear stress Read more
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