Kevin, I have been using CFSE to successfully label lymphocytes, you need very low concentrations. I have not attempted the cell division aspect, but I do know it is not composed of clean peaks. This is because the cells are not synchronized in most cases, and even if they started out in synchrony, they would not stay that way. There are papers out there, try looking for Lisa Geiselhart. She presented some good data at one of our Chesapeake Cytometry meetings in the fall. Good luck, Randy Fischer TherImmune Research Corporation 9700 Great Seneca Hwy Rockville, MD 20850 (240) 453-6256 RFischer@therimmune.com > ---------- > From: Kevin Waddick > Reply To: Kevin G Waddick > Sent: Monday, February 7, 2000 12:51 PM > To: Cytometry Mailing List > Subject: CFSE > > <<Message: Re: CFSE>> > The following message was originally sent to Gang Chen in response > to his thanking people who responded to his message entitled "Ask for > help". He was asking for help using CFSE on lymphocytes. Although I > sent the message twice, originally on 1/28/99, I never got a response. > > So, I have decided to go to the source of his help. Namely, the > correspondents to Cytometry e-mail. I hope that those who helped him > can also help me. > I didn't mention in my original message that I found concentrations > of CFSE as low as 1 ug/ml to be apparently highly toxic to the cell > lines when used for 10 minutes in the original loading of the cells. > > Kevin G. Waddick, Ph.D. > Parker Hughes Institute > 2657 Patton Road > St. Paul, MN 55113 > (651) 628-9988 x 224 > > > > Kevin Waddick wrote: > > I don't know why, but I haven't picked up on the exchange of > messages > regarding CFSE that you are thanking people for. I have been > trying to > use CFSE without success to analyze cell divisions. We have used > cell > lines from the B cell (Nalm-6 and Keller-2) and myeloid (HL-60) > lineages > and do not get any separable peaks that would reliably > correspond to > cell divisions. Is CFSE only useful in tracking cell divisions > when > starting with primary cells taken from a person or animal that > are then > prompted to enter cycle from the resting state? > I hope the responses you got can help me solve the problem I > am > having when trying to track divisions of cell line cells. Thanks > for any > help you can provide me. > > Dr. Kevin G. Waddick > Hughes Institute > 2657 Patton Road > St. Paul, MN 55113 > (651) 628-9988 x 224 > > > > >
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:06 EST