Introduction
Vena8 Fluoro+ biochips contain 8 parallel enclosed microcapillaries for continuous flow cell-based assays. Each micro capillary may be coated with a different adhesion molecule. Cell suspensions may then be injected using Cellix’s microfluidic pumps which supports a range of shear stresses for dynamic flow-based assays. Vena8 Fluoro+ biochips are particularly suited for applications requiring fluorescent immunostaining or confocal microscopy observation combined with flow-based experiments. Vena8 Fluoro+ biochips are supplied in packs of 10, facilitating 80 experiments per pack.
Vena8 Fluoro+ Features, Performance and Technical Specifications:
20x, 40x, 60x, 100x short working distance magnification microscopy; 60x, 100x oil-immersion microscopy.
Compatible with brightfield / phase contrast / fluorescent / confocal microscopy.
Low level fluorescence imaging, low fluorescent background.
Suitable for a wide range of cell suspensions and whole blood.
Easy to coat microcapillaries with a range of different adhesion molecules.
Biochip plastic is optically clear permitting detailed microscopy studies.
0.05–450 dyne/cm2 shear stresses easily obtained and controlled by the Mirus Evo Nanopump, ExiGo, UniGo and 4U microfluidic pumps.
Real time imaging under flow conditions.
*Considering human whole blood with a viscosity of 4.5 cP.
**Given for the flow of distilled water in a microcapillary with dimensions: 400 μm (W) x 100 μm (D) x 20 mm (L).
Vena8 Fluoro+ Biochip, Protocol #1: coating Vena8 Fluoro+ biochips
Step 1:
Cellix Vena8 Fluoro+ biochip is coated using a standard yellow tip pipette, by dispensing approximately 10 μL of protein (e.g. VCAM, ICAM, etc.) into each microchannel. Note the excess of liquid on the entrance and exit ports.
Step 2:
The Vena8 Fluoro+ biochip is then placed in a humidified box and sealed for 2 hours at room temperature or alternatively overnight at 4°C.
Step 3:
After the incubation period, add 10 μL of 0.1% BSA into each channel to ensure specificity of binding during the adhesion assay. The biochip is kept in the humidified box for a further 30 minutes.
Vena8 Fluoro+ Biochip Protocol #2: executing cell rolling, adhesion and migration assays under shear flow with Vena8 Fluoro+ biochips (manual version — not with VenaFlux platform)
Step 1:
Suspension cells (e.g. T cells, neutrophils, eosinophils, platelets) are re-suspended in culture medium at an appropriate concentration (typically 2–5 x 106/mL) in an Eppendorf tube.
Step 2:
Using the Cellix Mirus Evo nanopump or the ExiGo pump, 10 μL of media is dispensed from pump output cable. Following this, the output cable is inserted into a specified channel on the Vena8 Fluoro+ biochip.
Step 3:
Then using the Cellix Mirus Evo nanopump, 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 ligand and BSA used during coating. The waste comes out on the other side of the biochip and is collected inside the biochip microwell.
Step 4:
The waste is aspirated from the microwell of the Vena8 Fluoro+ biochip with a pipette.
Step 5:
Cell sample is placed into the microwell of this channel on the Vena8 Fluoro+ biochip.
Step 6:
Cells are introduced into the channel, by specifying the desired shear stress using VenaFlowAssay software or SmartFlo. The flow rate will be automatically calculated.
Step 7:
At each particular shear stress value, it is recommended that images of 3–5 fields of view of cell rolling and adhesion are acquired along the length of the channel.
So now you know the features and benefits of our Vena8 Fluoro+ and how they works. But, if you still have questions or want to learn more, check out our website, at wearecellix.com, or contact us at info@wearecellix.com.