E-Book Overview
In addition to providing chapters on the theoretical and practical aspects of flow cytometry, this book contains protocols used specifically for immediate use in the diagnostic and experimental measurements enabled by modern flow cytometric instrumentation. Research protocols cover cell cycle analysis and regulation, detection of rare cells, solid tumor DNA techniques, ploidy analysis and the application of flow cytometric methods for measurements of leukemias, necrosis, apoptosis, heat shock proteins, liver and bladder cancer, disease monitoring, and intracellular measurements. Hematologic and immunologic applications are also described by researchers with a particular expertise in their use. A special section is included for biotechnologists using flow cytometry in the study of protein secretion, the monitoring of fermentation processes, monitoring bioreactors, cell-cell electroporation, quantitation electrofusion, and electrochemotherapy.
E-Book Content
1 Basics of Flow Cytometry Gilbert Radcliff and Mark J. Jaroszeski 1. Introduction Flow cytometry is a laser-based technology that is used to measure characteristics of biological particles. This technology is used to perform measurements on whole cells as well as prepared cellular constttuents such as nuclei and organelles. Flow cytometers scan single particles or cells as they flow in a liquid medium past an excttation light source. The underlying princtple of flow cytometry is that light is scattered and fluorescence IS emttted as light from the excitation source strikes the moving particles. Light scattering and fluorescence is measured for each individual particle that passesthe excitation source. Scattering and emission data can be used to examine a variety of biochemical, biophysical, and molecular aspectsof partrcles. This unique and powerful technology is an important tool for many scientific dtsciplmes because i