Don't forget too that lipid transfection products are toxic to cells at
"relatively" high concentrations. If the lipofectamine level is too high,
cells might be dying off, or at least unhealthy, and could logically cause high
autofluorescence. What size plates vs what volume of cells are you using? Also how
confluent are the cells when transfected?
>From: Simon_Q_Rice@sbphrd.com
>To: Cytometry Mailing List
>Subject: Re: Lipofectamine auto fluorescence
>Date: Thu, 6 Sep 2001 16:16:27 +0100
>
>
>
>Tami,
>Are you sure your autofluorescence is caused by the lipofectamine? Don't
>you normally wash off the DNA/lipofectamine complex at least 24 hours
>before analysis? If not I suggest you do and replace with fresh medium,
>then leave your cells at least 24/48 hours before analysis. The other
>thing you might try is resuspending your cells in a phenol-red free medium
>just prior to analysis. Phenol red can cause autofluorescence.
>
>I hope that's helpful.
>
>Kind regards
>
>Simon
>
>
>
>
>trosario@cellmate.cb.uga.edu on 31-Aug-2001 21:52
>
>
>
>To: cyto-inbox
>cc:
>Subject: Lipofectamine auto fluorescence
>
>
>
>We are transfecting COS 7 cells with a plasmid containing GFP using
>lipofectamine. We see a large amount of auto fluorescence from the
>lipofectamine alone in our controls. Has anyone else experienced
>this? If so, how can we remedy the situation.
>Thanks for your help,
>Tami Rosario
>Department of Cellular Biology
>University of Georgia
>
>
>
>
>
>
>