I am looking for a way to visualize plasma membrane in mouse airway secretory cells, in paraffin sections by immunofluorescence.
In other words, it has to be a sufficiently abundant in plasma membrane protein to delineate the contours of the membrane.
Any suggestions?
Thanks!
Hi Michael, you could try EpCAM. It is well studies and is used for detecting epithelial derived circulating tumor cells.
Steingrimur Stefansson
Thank you, Steingrimur, we tried. It worked but not great: I didn't get the continuous line for plasma membrane, just dots.
I know it worked well for our collaborators; but they use frozen sections.
Michael Tuvim, bummer. Focal adhesion proteins, gap junction proteins, etc, both intracellular and membrane bound are usually not evenly distributed on the cell surfaces in vivo.
Have you tried fluorescent phalloidin? It could delineate individual cells.
Steingrimur Stefansson
Will do, Steingrimur. I was hoping for something in the PM its, as you mentioned, but will take the next best..
Michael Tuvim I just remembered that you can stain cell surface proteins with fluorescent lectins like conA. It is nonspecific in that it stains all cells having glycoproteins in their PM.
Merging lectin staining with phalloidin and EpCAM could provide mightily cool images, especially if you can do confocal.
I am not sure how well these work in histology sections but the CellMask dyes stain plasma membranes. Although they're recommneded for live cells mainly in the protocol I have used them on fixed cells grown on glass or transwells. They come in various fluorphors .
- Badges
- Science method