simultaneous surface/DNA analysis
Dr. Howard Petrie (h-petrie@ski.mskcc.org)
Tue, 25 Apr 1995 17:37:47 -0500
We are currently seeking a protocol which will allow us to do three color
surface immunophenotyping together with DNA content analysis. We can
already do the simultaneous surface staining/DNA staining quite nicely, but
only in two-color surface combinations. We have a FACStarPlus with dual
lasers
currently set at 488 and 600 nanometers. Since we routinely need this
configuration for FITC, PE, APC, and Texas-Red in various combinations for cell
sorting, it would really be best if we didn't have to swap out the dye head to
use UV (for Hoescht, etc.). In other words, we'd really like to find a
fluorochrome combination which will work using the existing beams. We've tried
FITC and PE (laser 1) with Texas-Red and 7-AAD (laser 2), but can't seem to
find a working combination of voltages and compensations on the latter. Since
7-AAD is very bright at 600nm excitation, we can't put this on laser 1 because
we still can't compensate it out of the Texas-Red due to the software
limitations. My understanding and previous experience suggests that we'll have
similar problems using PI. Any suggestions would be much appreciated.
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
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If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
