Fixation of HEp-2 in immunofluorescence slides
Posted 13 October 2010 - 05:58 AM
Anyone knows of any preservative or fixative which allows me this?
I use the following protocol:
Grow 3 ml of Hep-2 cells in 12 ml of DMEM. Incubated in stove of CO2 at 37 degrees.
I released the cells with trypsin and trasfer to other two bottles.
Then, I apply 50 ul of the HEp-2 cell in the slides,and Incubus in stove of CO2 at 37 degrees by 37 to 24 hours. Remove of the stove and fixed with methanol and acetone 1:1.
Posted 13 October 2010 - 08:27 AM
Posted 13 October 2010 - 11:16 AM
My problem is that after fixation with methanol/acetone at -20 degrees, the cells in the slides degrade when stored in the refrigerator, keeping only its stability in the freezer. But I need it to remain stable in the refrigerator.
How do that other treatment with paraformaldehyde you mentioned?
Posted 13 October 2010 - 12:02 PM
Posted 14 October 2010 - 06:47 AM
So, I'll try to use the collagen-coated coverglass and if it don't work I'll try to change the fixation.
Thank you very much for your help!
I await your return!
Posted 14 October 2010 - 07:32 AM