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IHC How Much Per Slide?


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5 replies to this topic

#1 EAG

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Posted 26 February 2012 - 10:16 AM

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?

Thanks!

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#2 bob1

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Posted 26 February 2012 - 04:20 PM

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.

#3 EAG

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Posted 26 February 2012 - 04:31 PM

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

#4 Rsm

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Posted 27 February 2012 - 08:03 AM

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.
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#5 EAG

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Posted 27 February 2012 - 09:23 AM

Thanks!!
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#6 ravibiosciences

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Posted 27 February 2012 - 01:13 PM

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.




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