Chapter 32 On-Line Flow Cytometry: A Versatile Method for Kinetic Measurements

Publisher Summary This chapter gives a technical description of the flow cytometer used to perform real-time kinetic studies, including the software necessary to acquire and analyze data. The chapter describes two applications of on-line flow cytometry (FCM). First, the real-time analysis of chromatin structure changes by enzymatic digestion of the DNA is shown. Changes in the fluorescence intensity of bound DNA stains occurred within minutes. These experiments therefore demonstrate the applicability of on-line flow cytometry to study fast kinetic changes. Second, an assay is described for measuring uptake kinetics of anthracycline drugs. Uptake profiles and changes therein after addition of specific membrane transport inhibitors can show the presence of typical multidrug-resistant cells. This assay is much more accurate and sensitive than classical FCM.