Author Topic: Tissue shredded after peroxidase IHC  (Read 106 times)

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Offline MAmanda

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Tissue shredded after peroxidase IHC
« on: August 22, 2018, 03:50:37 PM »
Quick question. Why would brain slices be shredded after peroxidase IHC?

Two sets of tissue, both perfused (4%PFA) by different people. Both sets sectioned on cryostat by different people. Kept in antifreeze.
Both ran through peroxidase IHC in the exact same manner.
TBS (1x) washes, methanolic peroxide wash (15 min), TBS+ blocking wash. Overnight Primary antibody. Then washes, secondary, tertiary streptavadin incubation. DAB for blue-black reactivity.

One set of tissue was "shredded" at the end of the IHC. the other remained intact remarkably well. Trying to find out what could be compromising the structural integrity of the tissue either during perfusion or sectioning on the cryostat, as those are the only 2 times the tissue were treated differently. Thank you in advance for any comments

Tissue shredded after peroxidase IHC
« on: August 22, 2018, 03:50:37 PM »