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>Subnanosecond polarized microfluorimetry in the time domain: An instrument for studying receptor trafficking in live cells
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Subnanosecond polarized microfluorimetry in the time domain: An instrument for studying receptor trafficking in live cells
We describe an instrument designed-to monitor molecular motions in multiphasic, weakly fluorescent microscopic systems. It combines synchrotron radiation, a low irradiance polarized microfluorimeter, and an automated, multiframing, single-photon-counting data acquisition system, and is capable of continually accumulating subnanosecond resolved anisotropy decays with a real-time resolution of about 60 s. The instrument has initially been built to monitor ligand-receptor interactions in living cells, but can equally be applied to the continual measurement of any dynamic process involving fluorescent molecules, that occurs over a time scale from a few minutes to several hours. As a particularly demanding demonstration of its capabilities, we have used it to monitor the environmental constraints imposed on the peptide hormone epidermal growth factor during its endocytosis and recycling to the cell surface in live cells. # 1998 American Institute of Physics. S0034-6748(98)03701-0
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