Abstract
Responses of single cells to stimuli are often heterogeneous. Bulk measurements by conventional biochemical and physiological techniques may fail to represent accurately the magnitude and time course of individual cell changes. Spatial heterogeneity of responses within single cells also occurs. For this reason, microscopic techniques with good three-dimensional resolution are needed to study individual cells as they respond to imposed stimuli and stresses. Increasingly, confocal microscopy of parameter-specific fluorophores is permitting direct observation of single-cell physiology with high spatial and temporal resolution. Confocal microscopy has become an essential tool to study the physiology of single living cells. As more parameter-indicating fluorophores are discovered, the range of applications of confocal microscopy can only increase. Uniquely, confocal microscopy permits observation of the physiology and metabolism of single organelles inside cells, as illustrated here for measurements of Ca 2+ , pH, ΔΨ, and membrane permeability of mitochondria. Overall, the impact of confocal microscopy on experimental cell physiology may someday compete with that of single-cell electrical recording.
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