Re: Mib-1 staining on wax sections of decalcified bone, adhesion problems after microwave antigen retrieval step

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From:Amos Brooks <>
To:"Mendes, Maria" <>, histonet <>
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    A few suggestions which may or may not work. First, when cutting the block,
if the section stays on the float bath for too long (usually in an attempt to
eliminate a wrinkle) the sections will fall off. Next we noticed a sharp decline
in fall offs when we stopped microwaveing and began steaming everything. You may
also have better luck using a more dilute H2O2 for longer time. When it comes to
slide adhesives it is usually best not to bother at all (especially if you can
get some sailinized slides or APES). I feel for you, we were having the same
problem with AE1/AE3 on bone marrows.
Amos Brooks

"Mendes, Maria" wrote:

> Hi I hope someone out there can help me with my problem. In our lab we are
> trying to perform immunohistochemistry on decalcified bone, mainly in
> respect to Mib-1. The problem is that after performing the antigen retrieval
> that involves putting sections in the microwave the sections fall off. Below
> are all the different things that have been tried to get around this
> problem. I would appreciate any help that could be sent my way. I may be
> contacted directly at <> .
> The following procedures have been tried with no success thus far:
> 1)      Contacted vendor that supplies Mib-1 to see if they had any
> information as well I did  a search on Pubmed to find any articles on the
> subject
> 2)      Used positively charged slides from two different vendors
> 3)      Used positively charged slides with albumin and then using a wet
> filter paper rolled over the section to get any air out of the section
> 4)      Used gelatinized slides with and without albumin and with and
> without the rolling feature above
> On all slides the wax was melted on the hot plate and the slides left
> overnight in a 37oC oven in some cases the slides were left over the
> weekend.
> Thanks,
> Maria (Toronto, Canada)
> Thanks,
> Maria Toronto, Canada

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