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Hi Michelle 
We imunostain the cells prior to fixation. With the modification of using HBSS 
insted of PBS prior to fixation but other wise we use the same protocol as 
for normal imunofluorescence. 

Ph.D. Stud. Ricardo Figueroa
Södertörns University

Tel: 08-6084732
Mob: 073-5631008
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On Tuesday 06 July 2004 10.58, Michelle Peckham wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear list,
>
> Does anyone have a method of fixing cells for immunofluroescence studies
> which will not cause permeabilization of the plasma membrane.  We have
> tried a variety of paraformaldehyde concentrations but still get a high
> % of permeabilised cells.
>
> 1% ParaF diluted direct into medium gives best results, but a lot of
> cells still get permeabilised.
>
> Thanks in advance
>
> Michelle
>
> Dr Michelle Peckham
> School of Biomedical Sciences
> Worsley Building
> University of Leeds
> Leeds
> LS2 9JT
> UK
>
> telephone: 0113 343 4348
> fax:       0113 343 4228
> website:   http://www.cell-motility.com