From: smmcgurran@mmm.com
Date: Mon Dec 15 2003 - 17:54:05 EST
Phil, I have done many experiments looking at whole mouse lung tissue (tumor bearing mice) by FACS. I process the whole mouse lung in about 3-4 ml of Complete Tumor Media (CTM) and make a single cell suspension. I use a ground glass tissue grinding system to "mash up" the entire mouse lung by hand and QS up to 20 ml with CTM in a 30 ml polypropylene centrifuge tube. Next, I centrifuge the tissue homogenate at 1500-1800 RPM for 10 min @ 15 degrees Celsius. I pour off the supernatant and resuspend the cell pellet in 2-3 ml of CTM. I take the resuspended tissue homogenate and plate out the cells in duplicate or triplicate @ 100 ul / well (depending on the degree of tumor growth for my studies). Once the cells are plated out I go through my antibody staining protocol for FACS. Usually at dilution of 1: 400. Hope this helps, Sean Sean M. McGurran 3M Pharmaceuticals-Pharmacology Bldg 270-2S-06 Tel: (651) 737-0843 Fax: (651) 737-5886 smmcgurran@mmm.com Christie Doxsee To: smmcgurran@mmm.com 12/15/2003 02:43 cc: PM Subject: FACS on lungs Thanks, Sean! Christie Doxsee Immunologist Drug Discovery, Pharmacology Pharmaceutical Division 3M Center, Bldg. 270-2S-06 St. Paul, MN 55144 651-733-4906 fax: 651-737-5886 cdoxsee@mmm.com ----- Forwarded by Christie Doxsee/US-Corporate/3M/US on 12/15/2003 02:43 PM ----- "Barren, Phil" <BarrenP@MedImmune To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> .com> cc: Subject: FACS on lungs 12/15/2003 09:53 AM 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
This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:44:06 EST
