How to quench fluorescence of Rhodamine labeled PE? - (Jun/26/2007 )

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hi, does anyone who has experiences on how to quench fluorescence of Rhodamine?I want the fluoresence totally comes from inside the cells not from the plasma membane. I know lowering culture medium PH can quench FITC of plasma membrane.

Thanks!!

-cell cracker-

Rho is not pH sensitive, so you can not do this trick. Is this a labeled lipid?

-genehunter-1-

QUOTE (genehunter-1 @ Jun 27 2007, 01:30 AM)

Rho is not pH sensitive, so you can not do this trick. Is this a labeled lipid?

yes，it is a labeled lipid,DHPE(Lissamine rhodamine B 1,2-dihexadecanoyl-sn-glycerol-3-phosphoethanolamine,triethylammonium salt(Rhdamine DHPE),it can exchange with the PE of plasma membrane, and can be endocytosed into a cell.I want to quench the fluorescence from plasma mem.

Thank you!

-cell cracker-

1)If you add enough of free unlabeled liposome, you maybe able to extract it off cell membrane, though this may not be 100%. 2) The alternative will be to use Fluorescein-lipid and CoCl2 quenching; NBD-lipid and dithionite is another good pair. However, all these approaches can only allow you to remove the ones that facing out (out leaflet). You will still have ~ 1/2 of the PE that is on the inner leaflet that can not be reached, neutralized/extracted.

If you are dealing with dead/fixed cells, limited detergent extraction may work as well.

-genehunter-1-

QUOTE (genehunter-1 @ Jun 27 2007, 10:08 PM)

THANK YOU! I Think I've got a solution.

-cell cracker-