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Fixation of cereals grain sections

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Paula Sampaio Paula Sampaio
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Fixation of cereals grain sections

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello all,

I have a collaborator that need to stain microtubules in multiple structures of coenocyte,  syncytium and cellularizing endosperm of early cereals grains. Do someone could give us some tips about a good method to fix and stain this kind of tissue sections for visualization of microtubules in confocal microscopy?

Thank you very much.

Best regards,

Paula





*********************************************************
Paula Sampaio, PhD.
IBMC - Instituto de Biologia Molecular e Celular
Universidade do Porto
Rua Campo Alegre 823
P4150-180 PORTO
PORTUGAL

Tel:  +351 226074900  Ext: 1606
Fax: +351 226099157


*********************************************************
Tobias Baskin Tobias Baskin
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Re: Fixation of cereals grain sections

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Re: Fixation of cereals grain sections

Hi,
        Check papers by R.C. Brown and B.E. Lemmon. THese two have published quite a few papers on this. Their protocol (s) would be a great place to start.

        Tobias

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello all,

I have a collaborator that need to stain microtubules in multiple structures of coenocyte,  syncytium and cellularizing endosperm of early cereals grains. Do someone could give us some tips about a good method to fix and stain this kind of tissue sections for visualization of microtubules in confocal microscopy?

Thank you very much.

Best regards,

Paula



*********************************************************
Paula Sampaio, PhD.
IBMC - Instituto de Biologia Molecular e Celular
Universidade do Porto
Rua Campo Alegre 823
P4150-180 PORTO
PORTUGAL

Tel:  +351 226074900  Ext: 1606
Fax: +351 226099157

email: [hidden email]

Web page: http://www.ibmc.up.pt/
*********************************************************


--
      _      ____          __   ____   
     /  \   /          / \    /   \ \        Tobias I. Baskin
    /   /  /          /   \   \      \         Biology Department
   /_ /   __      /__ \   \       \__    611 N. Pleasant St.
  /      /          /       \   \       \        University of Massachusetts
 /      /          /         \   \       \          Amherst, MA, 01003
/      / ___   /           \   \__/  \ ____
http://www.bio.umass.edu/biology/baskin/
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
Stanislav Vitha Stanislav Vitha
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Re: Fixation of cereals grain sections

In reply to this post by Paula Sampaio
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi,
also look at earlier papers by F. Baluska, he has been using Brown's
method with Steedman's wax embedding for immunofluorescence localization
of tubulin, actin  and a large number of other proteins in plants (mostly
in roots, but should work for kernels, too).
Baluska, F., Parker, J.S. and Barlow, P.W. (1992) Specific patterns of
cortical and endoplasmic microtubules associated with cell growth and
tissue differentiation in roots of maize (Zea mays L.). J. CellSci. 103:
191-200.

If you decide to use embedding in Steedman's wax, I have a detailed
protocol on our web site - the fixation is for actin, but the embedding
procedure is the same:

http://microscopy.tamu.edu/lab-protocols/Steedman_wax.pdf

Stan

On Fri, 31 Aug 2007 13:24:58 -0400, Tobias Baskin <[hidden email]>
wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>
>Hi,
> Check papers by R.C. Brown and B.E. Lemmon. THese two have
>published quite a few papers on this. Their protocol (s) would be a
>great place to start.
>
> Tobias
>
>>Search the CONFOCAL archive at
>>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello all,
>>
>>I have a collaborator that need to stain microtubules in multiple
>>structures of coenocyte,  syncytium and cellularizing endosperm of
>>early cereals grains. Do someone could give us some tips about a
>>good method to fix and stain this kind of tissue sections for
>>visualization of microtubules in confocal microscopy?
>>
>>Thank you very much.
>>
>>Best regards,
>>
>>Paula
>>
>>
>>
>>
>>*********************************************************
>>Paula Sampaio, PhD.
>>IBMC - Instituto de Biologia Molecular e Celular
>>Universidade do Porto
>>Rua Campo Alegre 823
>>P4150-180 PORTO
>>PORTUGAL
>>
>>Tel:  +351 226074900  Ext: 1606
>>Fax: +351 226099157
>>
>>email: [hidden email]
>>
>>Web page: http://www.ibmc.up.pt/
>>*********************************************************
>
>
>--
>       _      ____          __   ____
>      /  \   /          / \    /   \ \        Tobias I. Baskin
>     /   /  /          /   \   \      \         Biology Department
>    /_ /   __      /__ \   \       \__    611 N. Pleasant St.
>   /      /          /       \   \       \        University of
Massachusetts
>  /      /          /         \   \       \    Amherst, MA, 01003
>/      / ___   /           \   \__/  \ ____
>http://www.bio.umass.edu/biology/baskin/
>Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
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