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

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From:"a i d a n s c h u r r" <Aidan.Schurr@hvh.co.nz>
To:"Mendes, Maria" <mmendes@mtsinai.on.ca>, histonet@pathology.swmed.edu
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Hi Maria,

You don't make any mention of having tried silanised slides...  
We coat our own with 3-aminotriethoxysilane (APES), and don't 
have too many problems with things falling off.  Difficult tissues 
often need a lot longer to dry on - 3-4 days @37 degress won't go 
amiss.  One trick that can help is to draw water from under the 
section with a bit of filter paper before putting it in the incubator (we 
don't put any of our immuno's anywhere near a hot plate - too much 
damage to antigens for our liking...)

Good luck!
Aidan



> 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 mmendes@mtsinai.on.ca <mailto:mmendes@mtsinai.on.ca> .
> 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
> 


___________________________________________________
shin: device for finding furniture in the dark...
___________________________________________________
a i d a n   c   s c h u r r 
     mlso,  histology department
      hutt valley health
       lower hutt, new zealand

     ph.  ++64 4 5709173
     fax  ++64 4 5709214
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