Nile red compensation

From: Adelina de la Jara Valido (adjara1@hotmail.com)
Date: Thu Aug 15 2002 - 08:01:41 EST


Hello dear all,

I am trying to measure neutral and polar lipids in microalgae on a
FACSCalibur flow cytometer equipped with an argon laser using the dye Nile
red. With phytoplankton cells I get yellow fluorescence (FL2) for neutral
lipids and red fluorescence (FL3) for polar lipids and chlorophyll. Looking
at my results I think I am probably getting some kind of antagonistic effect
between both, the polar fraction and the chlorophyll fluorescences, since
the red fluorescence produced by non stained cells (chlorophyll in control
samples) is higher than that one produced by stained cells (polar lipids +
chlorophyll).

My question is: is it possible to properly compensate FL2 and FL3 using
stained cells with the solvatochromic dye Nile red (since this single dye is
supposed to fluoresce in different channels depending on the chemical
environment)?, do you know about any other possibility?.

And a last question, does anyone know about a more specific dye for long
chain fatty acids that can be excited at 488 nm?

Thank to all of you for your invaluable help.


Adelina de la Jara
Centro de Algología Aplicada
Gran Canaria
Spain




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