How to conduct a thrombosis assay using Mirus Evo or ExiGo Pump?
INTRODUCTION
Cellix offers precision microfluidic pumps for a wide range of application. Key features and benefits of these microfluidic pumps include:
Pulse free syringe pumps
Single and multichannel control
Multiple independent channel pumping
Patented active flow control for accurate sample delivery
Simple, easy-to-use control via iPad, PC
Ideal for microfluidics, shear stress , precision mixing and cell culture studies.
MIRUS EVO PUMP
Mirus Evo pump: accurate shear stresses / flow rates for cell analysis
Mirus Evo is a patented, precision, microfluidic, 8-channel syringe pump for cell analysis under shear flow mimicking physiological flow in the human vasculature. Includes MultiFlow8, a manifold that enables the flow from the Mirus Evo to be split equally into 8 separate tubes to conduct 8 assays simultaneously in the Vena8 range of biochips, resulting in higher throughput with this 8-channel syringe pump. Mirus Evo is PC- controlled by VenaFluxAssay software.
Features :
Includes MultiFlow8 for precision flow splitting with equal flow rate in each channel
MultiFlow8 contains 8 valves which can be switched on/off independently
Higher throughput enabling 8 assays in parallel
Patented flow damper to decrease syringe pump pulses
Flow rate: 100nl/min - 20µl/min with 100µl syringes. Increased flow rates possible with larger size syringes.
Dead volume: -600 µl
Flow direction reversible
PC controlled via VenaFlux assay software
EXIGO PUMP
ExiGo pump is a microfluidic syringe pump controlled by an iPad or a PC with SmartFlo. Suitable for numerous microfluidic applications, precise multichannel mixing, electrophysiology, single cell analysis , analytical biochemistry and RNA/ DNA analysis.
Features:
Precise flow control with active feedback via plug-and-play flow sensor (optional add-on)
Flow rate: 10 nl/min - 13ml/min
Standard syringes: l 00 µL-5 ml
Wash mode or programmable perfusion mode (constant, ramp, step, sine) with reversible flow direction
Rapid flow change (ms range)
Excellent long-term flow stability
iPad or PC (LabVIEW, Matlab, Python etc.) control which can contro l/program up to 4 pump modules independently
Wi-Fi communication
Use standard tubing for connection to any microfluidic biochip
Thrombosis Assays with Mirus Evo or ExiGo Pump Protocol: to coat channels of Vena8 Fluoro+ biochip with collagen and to perform thrombosis experiment using Mirus Evo or ExiGo pump
STEP 1
Cellix’s Vena8 Flouro+ biochip is coated using a standard yellow tip pipette, by dispensing approximately 12 μL of fibrillar collagen (200 μg/mL, NYCOMED 1130630) into each channel. Note the excess of liquid on the entrance and exit ports.
STEP 2
The Vena8 Fluoro+ biochip is then placed in a humidified box, which should be placed at 4°C for overnight coating.
STEP 3
After the incubation period, add 10 μL of 10% BSA into each channel to ensure specificity of binding during the adhesion assay. The biochip is kept in the humidified box for a further 15 minutes at room temperature.
Performing Thrombosis Assays with Mirus Evo or ExiGo Pump
STEP 1
Add 40 μL of media/buffer into the reservoir of the biochip before connecting the out cable from the pump.
STEP 2
Connect one end of thrombosis cable to the output cable of Mirus Evo pump/ExiGo pump and other end to the biochip inlet cable.
STEP 3
Using the Cellix Mirus Evo or the ExiGo pump, 10 μL of media is dispensed from the pump output cable. Following this, the output cable is inserted into a specified channel on the Vena8 Fluoro+ biochip.
STEP 4
Then using the Cellix https://www.wearecellix.com/mirusevopumpMirus Evo or the ExiGo pump, 40 μL of the media is injected through the channel at a shear stress of 40 dynes/cm2. This is done to wash the biochip of excess collagen and BSA used during coating. The waste comes out on the other side of the biochip and is collected in an Eppendorf tube.
STEP 5
Sample (e.g. whole blood, PRP, etc.) is introduced into the channel, by specifying the desired shear stress using VenaFlowAssay software or SmartFlo. The flow rate will be automatically calculated.
STEP 6
Disconnect the tubing from the blood sample tube after infusion of sample. Add buffer, e.g. PBS (100 μL) into the reservoir using a pipette and pull (aspirate) the buffer at appropriate shear stress Using Mirus Evo pump or the ExiGo pump. This is to wash whole blood from the channel.
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