I have performed Micronucleus assay after treated adherent HeLa and A431 cells for specific time. The cells were trypsinized into single cell, centrifuged @ 500rpm for 5 min. Fixed with Carnoys Fixative (1:5, Acetic acid:methanol) x2. Aspirated cells on clean and pre-frosted slide. The cells were observed under microscope but it looks like debris, might be cells were not fixed properly and damage.
what will be the reason why cells not fixed?
In which step, i might did mistake?
This protocol works fine with suspension cell lines. but found incorrect with adherent cells.
Thank you,
Muhammad Sheeraz Ahmad
Fix cells with 70% EtOH in water before staining with 5% Giemsa
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Bruna Rodrigues
Dear Vabeiryureilai.
I always used 4% paraformaldeyde with great results for adherent cells.
Godd Luck!
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