paraformalduhyde fixation and membrane integrity

Tsunehisa Namba drnamba at
Sat Feb 15 07:44:43 EST 1997

In article <5e23jr$7ls at>, 
rgrant at wrote:

>Andrew Doherty (A.Doherty at 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.

Tsunehisa Namba
Department of Anesthesia, Kyoto University Hospital
Sakyo-ku, Kyoto 606-01, Japan
E-mail: drnamba at

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