Date: Mon Apr 14 2008 - 07:14:52 EDT
I haven't had a reply from BD. Does anyone have the files listed below? Would there be ethical/legal/copywrite issues in using these in training courses? John Michie +27 21 9389 539 +27 21 933 8886 083 449 2225 -----Original Message----- From: Michie, John, Dr <jm5@sun.ac.za> Sent: 03 April 2008 12:44 PM To: cyto-inbox Subject: FCS DNA data files from BD FACS Operator Course Workbook I am looking for the fcs files that are referred to in the BD FACS Operator Course Workbook (337452 Rev. A). I wish to use them in a ModFit LT training module that I present as part of our BD-SUFCU Flow Cytometry Training Courses http://academic.sun.ac.za/flowcytometry/courses/courses.htm - if that is acceptable to BD. They are: O DNAH001 O JY001 O PBMC O PBMCJY001 O COLON001 O COLON002 O COLON003 John Michie +27 21 9389 539 +27 21 933 8886 083 449 2225 -----Original Message----- From: Dennis J. Young [mailto:djyoung@ucsd.edu] Sent: 02 April 2008 01:35 AM To: cyto-inbox Subject: RE: Background shift using Annexin Leaving out Calcium from the staining buffer should prevent the real Annexin-V binding. Many users have found better results with 10 (yes, TEN) fold less Annexin than the standard protocol, and especially while ALSO using ten times more cells (faster sample rate)! The positive controls determine how to interpret the experiment ;) I've not yet see anyone try mixing unlabeled Annexin-V with the labeled, which should be a good way of keeping the positives on scale, while saturating the binding. A better assay on a FACSCalibur includes Annexin-V FITC, PI, and DiIC1(5) - using the "Fourth Color" Ex635/Em660 :) Dennis J. Young Moores UCSD Cancer Center Flow Cytometry 2317-2Q 3855 Health Sciences Drive #0803 La Jolla CA 92093-0803 Lab (858) 822-0407 FAX (858) 822-0403 http://cancer.ucsd.edu/flow/ ________________________________ From: Rossi Ralph [mailto:ralph.rossi@petermac.org] Sent: Tuesday, March 25, 2008 10:23 PM To: cyto-inbox Subject: Background shift using Annexin Dear all, I have consistently seen a background shift when using Annexin as a marker for Apoptosis. Some of the reasons we've come up with are 1) too high titre of antibody 2) rough handling when lifting adherent cell types 3) rough handling during staining /spinning Does the list have any thoughts on this ? thanks ralph Regards Ralph Rossi Flow Facility Manager Peter MacCallum Cancer Centre Melbourne , Australia Email: ralph.rossi@petermac.org Ph 96563747 , 96561955 This email (including any attachments) may contain confidential and/or legally privileged information and is intended only to be read or used by the addressee. If you are not the intended addressee, any use, distribution, disclosure or copying of this email is strictly prohibited. Confidentiality and legal privilege attached to this email (including any attachments) are not waived or lost by reason of its mistaken delivery to you. If you have received this email in error, please delete it and notify us immediately by telephone or email. Peter MacCallum Cancer Centre provides no guarantee that this transmission is free of virus or that it has not been intercepted or altered and will not be liable for any delay in its receipt.
