![]() ![]() Table 1 lists clinical applications and cellular characteristics that are commonly measured. Tandem dyes with internal fluorescence resonance energy transfer can create even longer wavelengths and more colors. Commonly used dyes include propidium iodide, phycoerythrin, and fluorescein, although many other dyes are available. The use of multiple fluorochromes, each with similar excitation wavelengths and different emission wavelengths (or “colors”), allows several cell properties to be measured simultaneously. Upon returning to their resting states, the fluorochromes emit light energy at higher wavelengths. When labeled cells are passed by a light source, the fluorescent molecules are excited to a higher energy state. Additionally, antibodies conjugated to fluorescent dyes can bind specific proteins on cell membranes or inside cells. Fluorescent dyes may bind or intercalate with different cellular components such as DNA or RNA. Physical properties, such as size (represented by forward angle light scatter) and internal complexity (represented by right-angle scatter) can resolve certain cell populations. General Principlesįlow cytometry measures optical and fluorescence characteristics of single cells (or any other particle, including nuclei, microorganisms, chromosome preparations, and latex beads). This review will describe the basic principles of flow cytometry and provide an overview of some applications to hematology. Characteristics that can be measured include cell size, cytoplasmic complexity, DNA or RNA content, and a wide range of membrane-bound and intracellular proteins. Flow cytometry is used for immunophenotyping of a variety of specimens, including whole blood, bone marrow, serous cavity fluids, cerebrospinal fluid, urine, and solid tissues. Contemporary flow cytometers are much smaller, less expensive, more user-friendly, and well suited for high-volume operation. Whereas in the past flow cytometers were found only in larger academic centers, advances in technology now make it possible for community hospitals to use this methodology. The information obtained is both qualitative and quantitative. Flow cytometry provides rapid analysis of multiple characteristics of single cells. ![]()
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