I am looking for a way to visualize the glycocalyx of live cells (preferably fluorescence by blue or red excitation). After googling, it appears that the standard method is to use periodic acid (which inserts between saccharides and forms a pair of aldehyde groups) and a Schiff's reagent (which binds aldehydes, which are otherwise not too common on the cell/tissue surface).
Can anyone point me to a Schiff's reagent (or a suitable alternative) that is of the fluorescent (not the typical absorptive) variety, that fluoresces either blue/green (ex/em) or red/deep red (ex/em)?
Glycocalyx can also be stained using certain lectins. You might want to check the publications from S. Reitsma, e.g.:
The endothelial glycocalyx: composition, functions, and visualization.
Reitsma S, Slaaf DW, Vink H, van Zandvoort MA, oude Egbrink MG.
Pflugers Arch. 2007 Jun;454(3):345-59. Epub 2007 Jan 26. Review.
PMID: 17256154 [PubMed - indexed for MEDLINE]
Good luck!
Thank you Timo. I found that the periodic acid stain and Schiff's reagent are not suitable for live cell applications, so I will definitely look into the lectins.
thank you Michael. I am still wondering if there is anything that does not get recycled into the cytoplasm with the lipids over time?
It depends on the application, but another option could be the use of Safranin. I enclosed a paper describing the use of it for glycocalyx staining.
Thanks for the suggestion Hadar. The paper suggests fitc-conA binds exopolysacharides in the glycocalyx-- which sounds like a good fit for what I want to do. Could you explain what the safranin is staining? I read that it stains cell nuclei, and also that it is used as a counterstain for gram staining (presumably then it stains peptidoglycan of both gram + and gram - bacteria?), but also can be used for cartilage and mucin. So it seems to do a lot of different things!?
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Physical Phenomena
- Luminescence
- Fluorescence