Fluorescent Probes in the Study of Nucleic Acids and Chromatin in Living Cells
Fluorescence analysis provides a sensitive, selective, nondegradative approach to the measurement of nucleic acids in intact cells. Fluorophores of high quantum yield can be detected in very low concentrations, and, by the analysis of fluorescence quantities such as intensity, spectra, polarization, and decay time, it is possible to obtain significant information about the environment of the fluorophore. The position and interaction of the fluorophore with the intact structures within a cell can be determined. Usually this is done after fixation of the cell, because this allows access through the cell membrane for the probe and other reagents. There are a number of fluorophores that complex selectively with nucleic acids, but few that complex only with DNA or RNA. Selectivity can be improved by enzymatic digestion of RNA or DNA before application of the probe. Although fixed cells offer these technical advantages, there are many reasons for probing cells that retain full integrity, metabolic capacity, and viability. Recent reports indicate that this is becoming possible, and it is hoped that this review will help to stimulate further activity in this promising field.
KeywordsEthidium Bromide Fluorescent Probe Acridine Orange Deoxyribonucleic Acid Synthesis Acridine Orange Fluorescence
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