Cell membrane staining

William Sands was1h at clinmed.gla.ac.uk
Fri Dec 1 07:10:19 EST 1995


receptors per
>cell using fluorescence microscopy. We have some slides of 8-16 micron thick
>prostate cancer tissue cut from originally frozen tissue block.
>We are going to use FITC conjugated secondary antibody to stain steroid
>receptors.
>Here we wonder whether some cell membrame stains such as
>Texas Red conjugated Concanavalin or CellTracker CM-DiI (Molecular Probes)
>will stain cell membrane bright enough to isolate each cell during
>image analysis. In our slides, there would be only strips of cut cell membrane
>to be stained.
>Anyone have some experience similar to what I describe here?
>Any suggestions would be appreciated!
>
>Desok Kim

I don't think  the techniques you suggest will work on sectioned material,
as the CM-DiI will also stain intracellular membranes, and the con A will
bind to intracellular glycoprotens (secretory vesicles?). Normally, these
proteins are too big or lipophilic to cross intact cell membranes, so they
are reasonably useful for labelling the membrane of intact cells. Since
your cells are sectioned, I suggest you use an antibody specific to the
surface of the cells you are interested in.

Billy




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