17 August 1994 Simultaneous analysis of multiple fluorescent probes in single cells by microspectroscopic imaging
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Abstract
A spectrograph based imaging system attached to a microscope is described for use in monitoring the fluorescence of multiple probes loaded into single cells. Movements of ions, important in regulating cell function, are interrelated under many conditions. Since we are interested in the mechanisms underlying coupling between ion movements and metabolism, we required a microscope based system to analyze these parameters at the level of a single cell. The spectral imaging system was implemented for two purposes. (1) To resolve heterogeneities which exist in the response of individual cells in a population, and within any given cell due to loading of dye into more than one subcellular compartment, and (2) to study the temporal relations between several physiological parameters from a single cell. Since spectral imaging resolves signal from multiple probes simultaneously, the concentrations of several ions or metabolic factors can be followed in a single cell. Moreover, spatial information is acquired along one axis, providing information from probes located in individual compartments. The spectral imaging microscope and some specific applications are described.
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Raul Martinez-Zaguilan, Linda S. Tompkins, Ronald M. Lynch, "Simultaneous analysis of multiple fluorescent probes in single cells by microspectroscopic imaging", Proc. SPIE 2137, Time-Resolved Laser Spectroscopy in Biochemistry IV, (17 August 1994); doi: 10.1117/12.182728; https://doi.org/10.1117/12.182728
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