PKH26 ex max 551 (& 512), emm 567.
If you have a Innova70C-Spectrum, it's closest line is at 531, but PKH has
a secondary absorption max near the 514 line. The 514 is brighter and may
excite better. How about a 560 or 570 LP for detection?
I would use biotinylated rbc's instead.
Hoffmann-Fezer G; Trastl C; Beisker W; Berg D; Obermaier J; Kessler W;
Mysliwietz J; Schumm M; Filser J; Thierfelder S.
Preclinical evaluation of biotin labeling for red cell survival testing.
Annals of Hematology, 1997 May, 74(5):231-8.
At 11:42 AM 3/15/01 -0800, you wrote:
>
>I have a researcher who would like to utilize PKH26 as a tracking dye
>for rabbit rbc's survival studies. I have gotten reasonable separation
>of positives and negatives using a "PE" filter setup (555DLP beam
>splitter and 580/30 band pass) with 488 excitation on a MoFlo. The
>researcher is worried about cell toxicity of the PKH26 and wants to use
>as little as possible in his staining procedure. In an attempt to
>maximize fluorescence intensity, I've tried to "tune" the dye head laser
>(INNOVA 70) on the MoFlo to maximum excitation for PKH26 (551 nm) but
>seem to overwhelm the detection PMT with its 580/30 band pass. Any
>suggestions or advice on filter setup or use of PKH26 in this scenario
>is much appreciated.
>
>
---Dennis
Dennis J. Young
Flow Cytometry Core Facility
University of California, San Diego
Internal Medicine Group, Bldg #4, Rm 126
9500 Gilman Drive
La Jolla CA 92093-0671
Mail:<<mailto:djyoung@ucsd.edu>>
WWW:<<http://cancer.ucsd.edu/SResources/flow.htm>>
Telephone:(858) 822-0407 FAX: (858) 822-0403
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