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Immunofluorescence on FNA smears archive

FNA thyroid

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#1 pokemona

pokemona

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Posted 03 April 2012 - 02:59 PM

Hello there,

I was wondering (new to the field) if anyone has any experience in reusing already stained+air dried FNA smears on (ordinary-no polylysine) slides from a pathologist's archive to perform immunofluorescence and look at the cells under the confocal.

I list the pathologist's staining protocol for you to have a better impression on the material's state:

1) Fine needle aspiration smearing on glass slides
2) 3min May Grunvald staining (this stain is diluted in MeOH)
3) Rinse in H2O
4) Giemsa stain (1:10) for 5min
5) Rinse in H2O
6) Air dry

There are slides from recent FNAs (month's old) up to 6 years back. Is any of this usable for immunostaining and if so which protocol would you recommend? I presume the MeOH step partly fixes the cells and air drying too but I am not too confident about this?

Any help will be much appreciated.


PS: how about RNA extraction?
PS2: there are plenty of unwanted/unlysed red blood cells on the smear too.. How can i get rid of them?





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