Distinguishing two populations of cells by TEM

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Distinguishing two populations of cells by TEM

Postby apgoncalves » Jul 23 2015 1:27 am

Hey.

I'm planning to do some TEM experiments but I'm facing a problem and have no idea how to solve it (well, I have one idea but that wouldn't be very feasible). I want to mix two populations of cells and analyze their morphology when they are communicating and fusing with each other. I was wondering if there's a way to label the cells previous to mixing in order to distinguish them under the electron microscope.

Any feedback is appreciated! Thanks!

:D
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Re: Distinguishing two populations of cells by TEM

Postby kmuirhead » Aug 06 2015 3:24 am

You might try PKH26, a fluorescent dye that is often used to monitor cell fusion (for example: Tajito et al., doi: 10.1242/​jcs.090886, February 1, 2012 J Cell Sci 125, 662-672). Grecchi and Malatesta recently reported that PKH26 can be converted to an electron dense form that retains its membrane association (doi: 10.4081/ejh.2014.2449, Eur J Histochem. 2014 Oct 22; 58(4): 2449). When using fluorescence, the two cell types can be distinguished based on the emitted wavelength. I don't know if PKH67 could also be converted to an electron dense form, but even if it could, it wouldn't necessarily be distinguishable from the PKH26 after conversion. You might be better off to try a label that localizes to a different place in cell type 2 (for example: nuclear labeling with BrdU or EdU; Ngwenya et al. doi: 10.1369/jhc.4A6605.2005
J Histochem Cytochem July 2005 vol. 53 no. 7 821-832) or immunogold staining (if one of your cell types has a unique antigen).

Good luck!
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Re: Distinguishing two populations of cells by TEM

Postby apgoncalves » Aug 06 2015 4:51 pm

kmuirhead,
Thank you, great ideas! I'll definitely look at those papers.
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