This review illustrates the application of microspectrofluorometric techniques (including fluorescence microphotolysis) to the study in real time of the direct and indirect (photosensitization) effects of light in localized areas of single living cells. The photodynamic therapy of cancer by porphyrins provides good examples of studies of photosensitized alterations of membrane structures and organelle functionality through the use of specific endogenous (i.e. NAD(P)H) or exorgenous flouorescence probes. In addition, it is shown that microinjection of key cellular metabolites of the Krebs’ cycle (malate) or the pentose phosphate pathway (6-phosphogluconate) makes possible to also probe the effect of UV or visible light on determining steps of cellular metabolism.
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