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- Endocytobiosis and ...
- Volume 14
- Total internal refl...
Total internal reflection fluorescence microscopy (TIRFM) of acridine orange in single cells
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129-136
- Zusammenfsg.
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A miniaturized condensor unit for variable-angle total internal reflection fluorescence microscopy (TIRFM) has been developed and used for studies of cultivated endothelial cells incubated with the organelle marker acridine orange (AO). Upon excitation by an evanescent electromagnetic field, the penetration depth of incident light could be varied between about 50 nm and more than 200 nm. TIRFM signals were compared with fluorescence from the whole cell. AO fluorescence arising from the cell nucleus, lysosomes and those parts of the cytoplasm which were in close proximity to the plasma membrane, were thus distinguished. The selectivity of the method was further improved by using spectral and temporal (nanosecond) resolution.
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