Date: Wed Nov 28 2007 - 20:32:39 EST
Rich, Compensation can be done once a day as long as the panel does NOT change. In addition to compensation of samples, these beads validate the effectiveness of the conjugates used in your test because the binding is independent of antibody avidity. Therefore, they should be run with each panel. I would, however, recommend an additional control run throughout the day to validate the instrument performance over time. These can be beads (not necessary CST beads), which must hit target channels, previously selected by your CST beads. SP On 11/27/07 7:10 PM, "Richard D. Schretzenmair" <rds@mail.med.upenn.edu> wrote: > Hello, > > Currently, we are validating a FACSCanto for clinical use. In trying to > develop standard protocols for the operation of this instrument, a question > arose about how often compensation is required? We had intended to compensate > daily, but Current Protocols says that compensation is required for every run > (in the compensation myths section). When you are compensating 6 colors, then > include each antibody conjugated to a tandem dye, your could easily exceed a > dozen tubes for each compensation run. In the era of tight hospital budgets, > the cost of antibodies, compensation beads (or compensation samples) and labor > needs to be considered. > > It is my understanding that compensation is dictated by the instrument setup > (filters, mirrors, PMT Voltages, lasers) and the specific fluorochromes used. > This machine is QC'ed using DiVa 6's CST software, which does > a maniacally thorough job at QC. So, if the instrument setup is not changed, > CST QC is passed and PMT voltages held constant, how often does compensation > really have to be performed? Samples are run immediately after staining. It > seems that doing compensation with each run (several times a day), would be > more likely to introduce human error, than to correct for any true changes in > compensation. > > Considering the factors effecting compensation, is it really required for > every run, or even daily, given the above circumstances? > > Rich > > > > > > > Richard D. Schretzenmair > > > > > > Flow Cytometry and Cell Sorting Shared Resource > > > Abramson Cancer Center > > > University of Pennsylvania School of Medicine > > > 297 John Morgan Bldg. > > > 3620 Hamilton Walk > > > Philadelphia, PA 19104-6082 > > > > > > Phone: 215-898-3528 > > > FAX: 215-898-4227 > > > > > > rds@mail.med.upenn.edu > > > > -- Stephen P. Perfetto, MS.,MT. (ASCP) Manager, Core Flow Cytometry Facility Vaccine Research Center, NIH Building 40 40 Convent Dr., Room 5507 Bethesda, MD 20892-3015 email: sperfetto@nih.gov Phone: (301) 594-8659 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the senderıs own and not expressly made on behalf of the NIAID by one of its representatives.
