TY - JOUR
T1 - Fluorescence methods for continuous monitoring of transport in cells and vesicles
AU - Eidelman, Ofer
AU - Ioav Cabantchik, Z.
PY - 1989/1/1
Y1 - 1989/1/1
N2 - Transport mechanisms in biological systems are studied by following the movement of physiologically relevant substrates (or substrate analogs) from one compartment to another. This flux can be traced by a variety of physical or chemical techniques. The most widely used approach for tracing fluxes across membrane-sealed compartments is following the movement of a radiolabeled substrate by the separation of compartments at various time points, sampling of their contents, and the measurement of a label in one of the compartments. As this method is discrete in nature, the amount of information it can yield is fragmentary and often not precise enough for a thorough kinetic evaluation because of sampling and counting errors. Fluorescence is a suitable alternative to the classic radiotracer method because (1) it has the requisite sensitivity and temporal resolution for following the movement of fluorescent substrates among relatively small compartments, (2) it allows the selection of a discrete area of a given compartment (cell, tissue, or medium) by using physical (e.g., optical focusing) or chemical (e.g., impermeant light quencher) means, and (3) it permits a continuous monitoring of transport by fluorescence (CMTF) without requiring the periodic separation of compartments as an obligatory step of the transport protocol.
AB - Transport mechanisms in biological systems are studied by following the movement of physiologically relevant substrates (or substrate analogs) from one compartment to another. This flux can be traced by a variety of physical or chemical techniques. The most widely used approach for tracing fluxes across membrane-sealed compartments is following the movement of a radiolabeled substrate by the separation of compartments at various time points, sampling of their contents, and the measurement of a label in one of the compartments. As this method is discrete in nature, the amount of information it can yield is fragmentary and often not precise enough for a thorough kinetic evaluation because of sampling and counting errors. Fluorescence is a suitable alternative to the classic radiotracer method because (1) it has the requisite sensitivity and temporal resolution for following the movement of fluorescent substrates among relatively small compartments, (2) it allows the selection of a discrete area of a given compartment (cell, tissue, or medium) by using physical (e.g., optical focusing) or chemical (e.g., impermeant light quencher) means, and (3) it permits a continuous monitoring of transport by fluorescence (CMTF) without requiring the periodic separation of compartments as an obligatory step of the transport protocol.
UR - http://www.scopus.com/inward/record.url?scp=0024400445&partnerID=8YFLogxK
U2 - 10.1016/S0076-6879(89)72012-7
DO - 10.1016/S0076-6879(89)72012-7
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C2 - 2747525
AN - SCOPUS:0024400445
SN - 0076-6879
VL - 172
SP - 122
EP - 135
JO - Methods in Enzymology
JF - Methods in Enzymology
IS - C
ER -