At first, when I culture cells I could obviously see cells in channels that moving through the channels, but after 24 hour there is no cell.
Hi Maryam,
There could be potential leakage in the microfluidic device. For observing that, once you seed the cells, immediately look in the microscope, where are all the cells flowing towards. It is most likely there would be a leakage and cells are being collected at some reservoir/outlet. If cells are being collected in the outlet, then seal the outlet with a pipette tip with cell culture media. This will ensure that there is equal pressure at the inlet (from where you are seeding the cells) and at the outlet (with a pipette tip filled with cell culture media). Then you may see cells seeded in the microchannel.
Ami Mehta
Dear Ami
Thank you for your response.
There is no leakage.
After cell seeding, I add media to the outlet to create the balance, but after 24 h there is no cell inside channels. All cells are accumulate in inlet.
This means that there is blockage in the channel or the microchannels are not hydrophilic enough. Can you share more about your microfluidic device. Is it PDMS-based or PMMA-based? To make the microchannels more hydrophilic, after exposing to plasma, add collagen immediately and do the cell culture. If you want to store the chips for a later use, add collagen to the chips and place the chips in a petri dish along with a lid of 15 ml falcon tube filled with autoclaved water. Seal the petri dish with paraffin film and place it in fridge at 4 deg.
If you have a different device, let me know, so we can troubleshoot together.
It is PDMS based. We exposed it to plasma and then attached a coverslip to it. After, coated channels with gelatin 0.5 percent for more than 2 day then seeding cells.
Why I should keep it in 4 degree, because for cell culture I put them in an incubator.
Maryam Ansari It is likely that the gelatin solution is being evaporated in the incubator when the chips are placed for 2 straight days. This results in drying of the channels, hence the cells are not adhering to the channel.
Place the chips along with a 15ml/50ml falcon tube lid containing autoclaved water. This is prevent the evaporation. Also, you do not need to keep the chips with gelatin coating for 2 days. The coating of microchannels can happen as quick as in 30 mins. Try reducing this time as well.
Thank you for your information.
I will try another time and share the results.
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