Greetings again everyone: FRET frets solved. Set up unstained and FITC [or PE] and CY5 controls with the diode laser on and collect CY5 signal in FL4. After individual controls are run, turn off the diode laser [eliminates FL4] and look for CY5 FRET in FL3 generated from the resonance energy of FITC or PE. FL3 is a 670LP in the FACScalibur so CY5 looks great when you get FRET from FITC or PE. Thanks to Alice Givan for reminding me to check this out. All my best, -Rich Richard F. Konz, Jr. Scientist Manager, Flow and Image Cytometry Facility Wyeth 200 Cambridge Park Drive Cambridge, MA 02140 USA Office: 617-665-5522 Lab: 617-665-5551 >>> "Richard Konz" <RKonz@wyeth.com> 07/01/02 01:55PM >>> Greetings everyone: An investigator in my research group is doing FRET by facs analysis on a two laser FACScalibur [FITC/PE and CY5]. During experimental setup we use the red diode laser to look at the CY5 staining signal. However when we do the FRET experiment we need to have the diode laser off. When you shut the diode laser off, Cell Quest will eliminate FL4 as a collection parameter. Thus far, we are unplugging the diode laser from the board while we are doing the FRET experiment. I could install a switch to cut the power, but in a multi-user facility this would probably cause problems with the users forgetting to turn it back on. Does anyone have any ideas or potential workarounds with this FRET combination? Thanks for your input and all my best regards, -Rich Richard F. Konz, Jr. Scientist Manager, Flow and Image Cytometry Facility Wyeth 200 Cambridge Park Drive Cambridge, MA 02140 USA Office: 617-665-5522 Lab: 617-665-5551
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:14 EST