[Histonet] Re: Please help with me--routine immunofluorescent staining on kidney specimens

[yichao wu]

Dear Dr. Callis,

Thank you very much! And could you kindly suggest me that what kind of 
FITC-labelled primary antibodies to IgG and IgA etc should be applied on 
renal frozen sections?

Are F(ab'2)  antibodies preferred than IgG fraction?

Is methanol caused the autofluorescence or unproper fluorescence in 
glomerular capillaries?

Thank you.

Yichao WU


>From: Gayle Callis <gcallis <@t> montana.edu>
>To: "yichao wu" <yichaowu <@t> hotmail.com>
>Subject: Re: Please help with me--routine immunofluorescent staining on  
>kidney specimens
>Date: Thu, 18 Mar 2004 09:15:08 -0700
>
>Get rid of methanol, use cold pure acetone fixation ONLY!!
>
> >Another questions to Dr. Callis, you have mentioned to me that,
> >"A cryomold is what you embed tissue in - to form a block.The chuck is 
>what
> >holds the block when cutting in cryostat.DO NOT USE THE CHUCK FOR SNAP
> >FREEZING..."
> >
> >I just wonder then, how do you TRANSFER the block from cryomold onto the
> >chuck?
>
>Pop the block out of mold and freeze block onto little metal chuck that
>holds blocks during sectioning.  Do this with OCT. Use your fingers and
>work quickly to not melt parts of the block where tissue is located.
>
>
>Gayle Callis
>MT,HT,HTL(ASCP)
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>
>

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