Annie, We have been playing with a number of different tracking dyes lately to use in conjunction with CFSE and that do not seem to cost as much as PKH-26. One that I have recently tried is the CM-DiI carbocyanin dye from molecular probes. In the one experiment I have run to date, I found that the cells can be visualized on a stock dual laser FACSCalibur providing that the cells are heavily loaded. The argon laser does excite the dye, and it shows strong fluoresnce in the FL2 and FL3 channels, leaving FL1 and FL4 clear for other fluors. -james James F. George, Ph.D. Associate Professor of Surgery University of Alabama at Birmingham Birmingham, Alabama 35294-0007 jgeorge@uab.edu > > Hi, > > Has anyone had flow experiences with Molecular Probes' product MitoTracker Red (CMXRos), > in particular with 3-colour staining? > > We are using a FACSCalibur with a 488 nm laser and find that the fluorescence of > CMXRos alone is detected in FL2, FL3 and FL4. We can decrease the FL2 signal to a > negative level using FL2 - 99.9% FL3 compensation while retaining the signal in FL3 > and FL4. However, we cannot decrease the signal in FL3 using FL3 - 99.9% FL2 (or FL4) > compensation. > > I am having a hard time conceptualizing this compensation model. Any ideas? > > Thanks so much. > > Annie Bang
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