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Help needed with IF staining.


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#1 pSmad

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Posted 21 September 2009 - 10:36 AM

Hi everyone,

I have previously done alot of immunofluorescence staining of brain sections and have recently transitioned from neurobiology to immunology. Now I'm required to work with spleen, liver and MLN. Due to the lack of experience in these organs I need some guidance. I have fresh, snap frozen spleen, liver and MLN in OCT

1) What is the best thickness to section these organs? 4,8 or 10 um?
2) do they need to be fixed immediately after sectioning?
3) how do I store these slides for longer time periods and still preserve the surface CDs for future staining?

Any advice and tips on these questions will be very much appreciated!

Thanks

Best,

pSmad

#2 SaraK

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Posted 06 November 2009 - 09:26 AM

I don't know how helpful this is but our lab sections spleens and lymph nodes at 8 um on to lysine treated slides and stores them in a slide box wrapped in plastic at -80 for long periods of time(years) we have not had any trouble with staining on older sections.

#3 miBunny

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Posted 06 November 2009 - 06:09 PM

I'll ditto Sara's answer for sectioning and storing. We do the fix after we thaw the sections.
btw, welcome to immunology!




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