From: Mara.Rocchi@mri.sari.ac.uk
Date: Mon Mar 17 2003 - 09:50:20 EST
Dear all, not exactly a flow question, but I know that the wisdom is great in this group and that many of you have started working at a microscope ..... I am working with 2 different cellular populations that are mixed in the same well and I need to recognise which one is which. Morphologically they look very similar. I would like to stain one of the two lines with a stain or a protocol that will not kill my cells, will be retained for at least a week and could be used at the light microscope, since I have to monitor their morphology day by day. I also have to count them both. I have tried CFSE and a fluorescent microscope, but by the time I count one population my fluorescence fades and then is very difficult to recognise real negatives from CFSE-faded cells. FACS is not much of help either, since the scatter profiles of my cells do overlap a lot and due to the nature of my cells (very variable in size) my CFSE stain goes from a very bright one to a very very dim one. Also I cannot sample my well every day (not enough cells). I know I am probably asking a lot, but can anybody out there help? Thanks a lot Mara ******************************* Dr. Mara Rocchi MVB Ph.D. Moredun Research Institute Pentland Science Park Bush Loan, Penicuik EH26 0PZ Scotland, UK Tel: +44-131-4455111 Fax: +44-131-4456111 e-mail: mara.rocchi@mri.sari.ac.uk The information contained in this e-mail message and any files transmitted with it is confidential and may be legally privileged. It is intended only for the addressee and others authorised to receive it. If you are not the intended recipient or the person responsible for delivering the message to the intended recipient you are advised that you have received the e-mail in error and that any disclosure, copying, distribution or action taken in reliance on the contents of the e-mail and its attachments is strictly prohibited and may be unlawful. Please advise the sender immediately by replying to this e-mail and delete it and all copies from your system.
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