Fixation of calcein stained cells
Edgar Rivedal (edgar.rivedal@labmed.uio.no)
Mon, 4 Nov 1996 14:28:54 +0100
Does anyone have experience with fixation of cells stained with Calcein-AM?
We fix epithelial cell lines stained with Calcein-AM in 4 % formalin.
Formalin is supposed to fix the calcein to the cells. What we see is a
relatively large drop in the calcein level over a few hours after fixation,
in spite of keeping the samples in the dark and on ice. What we do is to
detach the cells into 0.5 ml trypsin and mix this with 0.5 ml 8% formalin
and run this at the flow cytometer within some hours, i.e. we do not remove
the trypsin or formalin prior to analysis because the fixed cells seem to
dislike centrifugation.
________________________________________________________
Edgar Rivedal, Ph.D. (edgar.rivedal@labmed.uio.no)
Institute for Cancer Research
The Norwegian Radium Hospital tel. +47 22 93 47 00
N-0310 Oslo, Norway fax +47 22 93 57 67
________________________________________________________
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
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
