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Author Topic: Inconsistent staining on a slide  (Read 2539 times)

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Offline uchtmann

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Inconsistent staining on a slide
« on: November 23, 2015, 10:18:53 AM »
I'm consistently seeing inconsistent staining within a slide  :o
I have not noticed any patterns with this inconsistency (within a tissue or between sections, top/bottom, left/right, inside/outside, etc).  It is over different tissue types (prostate, liver, lung, bladder, etc.), different species (mouse, rat) and different antibodies (ERa, AR, BMP4, CYP11a1, etc.).

I've tried a lot of things to troubleshoot including:

Baking in 75C oven for 30mins or not baking at all
Fresh dehydration and hydration reagents
Making sure they are well-submerged when dipping
Making sure the tissues never dry out during any step
Making sure the antibodies are evenly coating the tissue
Fresh DAB that is evenly spread
Boiling or not boiling during antigen unmasking step in the microwave


Anyone else have experience with this or have any suggestions?
Thank you!
Kristen
« Last Edit: November 23, 2015, 12:56:07 PM by uchtmann »

Inconsistent staining on a slide
« on: November 23, 2015, 10:18:53 AM »

Offline gula

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Re: Inconsistent staining on a slide
« Reply #1 on: November 28, 2015, 01:57:48 PM »
For manual staining the Pap-Pen comes to my mind. If it is too near to the section, it may spread over the tissue-edges and inhibit staining.

gula

Re: Inconsistent staining on a slide
« Reply #1 on: November 28, 2015, 01:57:48 PM »