Dear Katja It is possible to use Pe Cy5. But compensation required is not only for the spill over but also due to the extra excitation of Cy5 caused by the 633 line. Choose for a few as possible different moabs conjugated with Pe Cy5. Let say in a B-cel panel only a CD19-Pe Cy5, and that combined with different Fitc, Pe and APC moab's. Because for every other moab conjugated with Pe Cy5 you have an altered spill-over. We have choosen for the use of Percp as FL3 on the FACSCalibur and waiting for the availability of Percp Cy5.5 coupled to moab's. The advantage is that you expect no extra excitation of Cy5.5 by the 633 line. So only compensation is required caused by the spill over. Greetings, Arie van der Marel Leiden University Medical Centre Section Flowcytometry E1-Q74 Albinusdreef 2 2333 ZA Leiden The Netherlands
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