I don't know the specificity or utility for suspended cells of this new assay but it sure looks easy and may distinguish live and dead eosinophils. I expect the excitation max should be a bit longer than 488 nm. J Immunol Methods 2002 Feb 1;260(1-2):273-8 A simple method for the in situ detection of eosinophils. Ain R, Tash JS, Soares MJ. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA. A simple and rapid method for identifying eosinophils in tissue sections is described. The assay is based on the incubation of frozen tissue sections with phenol red and the detection of cellular fluorescence. A one-to-one relationship was observed between cells exhibiting fluorescence following exposure to phenol red and eosinophils as identified by histochemical detection of eosinophil peroxidase (EPO) activity. Using the phenol red assay, eosinophils were detected in various eosinophil-infiltrated tissues, including uteri of rats from day 1 of pregnancy and uteri of prepubertal estrogen-treated rats. Intensity of the fluorescing eosinophils was dependent upon phenol red concentration and duration of incubation. The phenol red method of eosinophil identification was disrupted by co-incubation with resorcinol, an EPO inhibitor, or catalase, a hydrogen peroxide scavenger. The merits of this assay are its simplicity and compatibility with other procedures, such as histochemistry, immunocytochemistry, and in situ ligand-receptor binding. Karim Vermaelen wrote: > Hi everybody, > Can anyone recommend an accurate method to count eosinophils in rat > blood using FACS? Someone in our lab is currently doing manual counts > using eosin-treated whole blood... quite excruciating indeed > Thanks for any suggestions! > Karim Vermaelen > > Karim Y. Vermaelen, MD > Dept. of Respiratory Diseases > Ghent University Hospital 7K12ie > De Pintelaan 185 > B-9000 Ghent > > tel +32 9 240 2605 > fax +32 9 240 2625 > e-mail Karim.Vermaelen@rug.ac.be, karimv@mac.com
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