How to avoid salt crystallization in Immunocytochemistry?

Go to the profile of Chinmaya Sadangi
Jan 20, 2018
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I did an ICC and while imaging, I found that there are salt crystals on the coverslips. How can I avoid it?
My last steps are as follows:
After secondary, I wash the coverslips with PBS 3 times for 5 min.
I put a drop of my mounting media on a glass slide.
Take out the coverslip from the PBS, gently tap it on a kim wipe and place it on the glass slide.
Next, I keep the slides overnight at RT in dark and the following day I seal them using nail varnish.
Thanks

3 Comments

Go to the profile of mdfenko
mdfenko 5 months ago

are you performing the reactions on the coverslips?

if not, then why wash with pbs?

if so, then give a final rinse with either phosphate buffer (without salt) or water.

Go to the profile of Chinmaya Sadangi
Chinmaya Sadangi 5 months ago

Yes, I am doing it on coverslips. After secondary ab, washing 3x with PBS, should I do an additional wash with 3x with water?

Go to the profile of mdfenko
mdfenko 5 months ago

just a quick rinse should be sufficient to remove salt. you don't want to expose the antibodies to the water for a prolonged period. you could use phosphate buffer (the same concentration that you use in the pbs).