A whole blood alternative to traditional methods for CD4+ T lymphocyte determination

Flow cytometry, the method of choice for determining subset cell populations, requires sophisticated instrumentation and highly trained operators. An alternative method for subpopulation determinations is presented that is fast, simple, and readily interpreted for absolute subset counts. This method uses antibody labeled microspheres added to whole blood and after lysing, read on a hemocytometer using an ordinary light microscope. The total time of testing is less than 10 min with results comparable to that obtained using a flow cytometer and a hematology analyzer to calculate absolute subpopulation values. This method is demonstrated for CD4+ lymphocytes which have considerable value in following HIV-infected individuals.