I'm having a problem with gelatin coating. My gelatin for some reason solidifies prematurely and when i look under the microscope, i can see a lot of lumps. I tried many different ways from leaving longer in the warm bath, to vortex in between pipetting but none worked. I don't know what else can I do to be honest.
After pipetting the gelatin onto the coverslip, I leave them at room temp. (side a box to prevent bleaching) for 15 mins, and suck out the excess. Do you think any of these steps make it solidify unevenly?
Michael: The gelatin is made by other people in the lab and aliquot out in small tubes and kept in the freezer. So I only just need to take the tube out and warm it up to 37 degree tho.
It can be a bad batch or prepared not properly. Perhaps it is easier to prepare a fresh portion yourself.
Do you think it's also because I warm it up not enough? I leave the gel in water bath for quite a while however, but i'm not sure when i take the tube out to pipette onto my coverslip, during that time, the gelatin might just resolidify?
It can be everything. You have to check each step carefully. If it is already aliquoted, it might be not your fault. Heating to 50-60 degrees centigrade might remove the lumps. However, heating too high (boiling) might spoil it completely.
You may need freshly prepared gelatin to eliminate the error.
I've just learned that the gelatin was not made by people in the lab but was purchased from a company. So in theory, it should not be contaminated tho. What concentration do you use to coat the plate? Is 1mg/ml ok? Or you think it's too high?
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