IHC Cell Culture

From:bliven.laura@marshfieldclinic.org


In a previous email I requested info on staining West Nile Virus on cell cultures.  After receiving many valuable and educational emails, I have a few additional questions.

I was able to get (free from another dept.) some glass chambered slides that apparently have some type of weak adhesive or charge on them.  I've also decided to try cold acetone and methanol fixation (2 separate sets of slides).

So,
someone mentioned to make sure to let the slides dry out before plunging them into fixative, so the cells won't all come off the slides.  I've never let slides for IHC dry out.  Should I let the slides dry out or should I skip this part?

Also, some mentioned using detergents, like Triton X100, as it will facilitate the penetration of the IHC reagents through the cell wall, which makes perfect sense. But, some say to use it on all slides, others on only formaldehyde fixed slides (I'm using acetone and methanol fixed slides), and another mentioned that it shouldn't be used as it will dissolve the cells from the glass surface.

Thanks for any info.
Laura  




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