Hi, I have grown adherent cells on glass coverslip and would like to use them for Immuno Fluorescence experiment later. I would like to know if it is possible to dehydrate the cells gradually using different percentage of ethanol and store them for now, which I can fix later and use. I heard that fixing the cells and then storing them might lead to detachment of the cells if the fixed slides are more than a week old. Can someone help with the protocol, if any please?
In our lab, we use formaldehyde method to fix the cells. Procedure as follows,
1. Obtain 80-90% confluent cells in 24-well plate.
2. Wash cells twice with 1ml of 1XPBS.
3. Add 500ul of 3.7% formaldehyde in 1XPBS.
4. Incubate at RT for 10mins.
5. Wash with 1XPBS twice.
6. Add 500ul of 1XPBS with penstrep.
these can be stored at 4C for many months.
Hope this is helpful.
Thanking you.
- Thank you for the protocol Reshma. I heard that the cells would detach after a week or so, if they are fixed and stored. Can you please let me know if the above protocol worked for you and if the cells were intact even after weeks.
As you have added formaldehyde, it will fix the cells intact. they would not get detached. In our laboratory, we fix the cells in same way and use them even after 6 months.
hi, during your protocol, did you carry out dehydration with increasing concentration of cold ethanol following FA treatment?
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