From: Mara.Rocchi@mri.sari.ac.uk
Date: Tue Dec 16 2003 - 04:21:57 EST
Hello Phil, not exactly a mechanical dissociation approach but... we have recently collected lung cells from ovine lung tumours and normal ovine lungs after dissociation with Accutase. http://www.innovativecelltech.com/accutase.html There are others non-enzymatic treatments available from other companies if you don't want to purchase this one. We decided not to use trypsin since we did not know if our antigen was trypsin sensitive. If you decide to use this approach is mandatory that you use a dead cells excluding dye in your analysis, since forward and side scatter are not sufficient to identify damaged cells. Also we found that you need to pre-filter your sample if you don't want to clog your lines. We pre-filtered the sample BEFORE staining. Please let me know if you need a more detailed protocol and I'll be happy to send it to you. Could you also post a summary of the answers to the list? Thanks. Best regards and season greetings Mara ******************************* Dr. Mara Rocchi Ph.D. Flow Cytometer Manager 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 -----Original Message----- From: Barren, Phil [mailto:BarrenP@MedImmune.com] Sent: 15 December 2003 15:54 To: cyto-inbox Subject: FACS on lungs FLOWers Is anyone currently ( or in the past) done any Flow Cytometry on lung tissue ( Mouse, mechanical dissociation)?? Infiltrating immune cells and changes in response to challenge and treatment We are approaching this work for one of our models, and I was wondering if any one has any experience and pointers. Thank you for your time Philip Barren 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.
This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:44:06 EST
