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IHC How Much Per Slide? - (Feb/26/2012 )

I need to do IHC in 250 slides and I was wondering how much reagents per slide I need.

I need to considered how many microliters per slide?




It depends on the sample size (diameter in mm?) I have seen tissue samples that are1-2 mm across, but also samples that fill the whole slide. It will also depend on the techniques you plan on using and whether you are using an automated stainer or not.


The samples are small, something like 2-3mm and I will not use any automation


I usually use 100ul for a whole slide (and that's too much), so I'd use 10ul for a small sample (5ul should work as well). Cover with parafilm to prevent drying.




If you have time and your staining is not expensive :). It would be good to optimize your conditions from 5-100microL, careful and dont get the slide completely dry. More the dye you use more the background :(

Good luck.