paraformalduhyde fixation and membrane integrity

Tsunehisa Namba drnamba at kuhp.kyoto-u.ac.jp
Sat Feb 15 07:44:43 EST 1997


In article <5e23jr$7ls at news.ox.ac.uk>, 
rgrant at molbiol.ox.ac.uk.remove_this_bit wrote:

>Andrew Doherty (A.Doherty at Bris.ac.uk) wrote:
>} HI everyone - a simple question. When cells are fixed in 3%
>} paraformaldehyde, is the integrity of the membrane maintained? Could I
>} detect surface expressed receptors with antibodies without seeing the
>} intracellular pools?
>
>Um, practically, no the integrity is not maintained.  OK, so we always
>permeabilize as well as PFA to see intracellular antigens, but that's more
>to be absolutely sure they're full of holes.  I've looked at actin
>staining in PFA-alone fixed cells and seen it.  (actin, that is :-))

I think Richard used phalloidin as a probe for actin staining.  That's 
substantially smaller than Ig.  Once I have forgot to permeabilize the 
cells to see intracellular antigen.  I could not see anything in healthy 
cells.  Morbid cells were somewhat stained because the membrane integrity 
has already been damaged.
 Nam

-- 
***************************************************
Tsunehisa Namba
Department of Anesthesia, Kyoto University Hospital
Sakyo-ku, Kyoto 606-01, Japan
Fax:+81-75-752-3259
E-mail: drnamba at kuhp.kyoto-u.ac.jp



More information about the Methods mailing list