--- On Wed, 24 Mar 1999 10:00:56 +1100 Kristy Weir <cozens@postoffice.utas.edu.au> wrote: > >Dear Flow-ers > > I'm a student using Mark's email (Kristy Weir). >I'm using Immuno-4 subtraction program on a coulter elite. Is overton >subtraction/Immuno-4 used widely? How acceptable is it for publishing data? >I'm looking at sheep dendritic cells for phenotype analysis. Negative >controls are within the first log decade and are quite clean. I've compared >Immuno-4 to placing a gate manually and the results are quite comparable >(within 2-5% difference). >Any help would be much appreciated, thanks in advance. > Algorithms such as Oversub and Immuno are useful for phenotypic data whch has a continuous distributuion ie no clear positive population. For antigens where there is a clear positive and negative, like human CD4 and CD8, a region can just be drawn on the positives to get percentage or relative fluorescence intesntiy information. For antigens with a continuous distribution, such as HLADR in humans, it is much more difficult to determine the percentage of cells which actually express that molecule. A good way to handle data such as this is to describe the fluorescence intensity of the entire population. However, if you wish to quantitate the percent positve as well, I think using an algoritm is a superior method as compared to simply drawing a cursor. As I recall, the Immuno program requires the user to set certain regions, negative, positive, etc. Also, I think there are certain requirements that need to be met for dat a to be analyzabel by this program. The Oversub algorithm requires less user input, and might be more consistent. I think if you stick to one method, for comparative purposes this is OK. Good luck, Tom -------------------------------------------------------- Thomas W. Mc Closkey, Ph. D. Director, Flow Cytometry North Shore University Hospital Biomedical Research Center 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office]; 516-562-1135/4641 [lab] fax: 516-562-2866 3/26/99 5:02:23 PM E-mail: thomasm@nshs.edu --------------------------------------------------------
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