Lipids staining and/or foam cells detection

[MCOlivier]

Hi dear CyTOFers

Do any of you know about a technic/compound that could be used in CyTOF for intracellular lipid staining (aim is to characterize foam cells in mouse atherosclerosis) ?
Thank a lot and best regards !
Olivier

[mleipold]

Hi Olivier,

I'm not really that familiar with that type of cell.....how are they characterized in regular fluorescent flow?

For example, if they're detected using some kind of fluorescent tag, could you use the same probe and then detect with anti-fluorophore antibody? Sort of like how Zinaida Good adapted CFSE with anti-FITC-metal (PMID: 30742126).

Or do you need an actual metal-labeled lipid for some reason?


Mike

[MCOlivier]

Hi Mike

First of all, thanks for your answer and help !

To my knowledge, only BODIPY dies (http://mikrobiyolbul.org/managete/fu_fo ... 0Ergin.pdf) and LipidTOX dies (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4143162/) are used for neutral lipids quantification.

But sadly, those dies are not built with standard fluorochromes like FITC, then, not detectable with antibodies... this explains my demand of "metal labeled compound/reactive for lipid detection"

I also found that Osmium as well as Ruthenium tetroxides can be used to label cell membrane lipids, but these won be quntificative for droplets detection by Mass... (https://onlinelibrary.wiley.com/doi/epd ... to.a.22848)

I'm quite out of technological knowledge here... sadly
Best regards !
Olivier

[mleipold]

Hi Olivier,

I guess the tricky part is quantifying only the intracellular lipids, rather than the total lipids like the OsO4 or RuO4.

I did find this polyclonal anti-BODIPY antibody: https://www.thermofisher.com/antibody/p ... nal/A-5770 (RRID AB_2536193)

I don't think it's been used for flow, but might be worth a try?


There's also this thread on ResearchGate: https://www.researchgate.net/post/Which ... ts_in_cell


Mike