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 _________________________________________________________________ Charle con sus amigos online usando MSN Messenger: http://messenger.msn.com
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