A METHOD FOR OBTAINING STRUCTURAL AND FUNCTIONAL INFORMATION ON PROTEINS, BASED ON POLARIZATION FLUORESCENCE MICROSCOPY, AND A DEVICE IMPLEMENTING SAID METHOD

摘要

The invented method of acquiring structural and functional information on proteins based on polarization fluorescent microscopy is characterized in that protein with a bonded fluorophore is subjected to two-photon or multi-photon fluorescent microscopy where the investigated protein with fluorophore is irradiated by a laser beam the light of which is polarized in at least two different polarizations and a which excites fluorophore fluorescence. Based on information on localization, intensities and polarization of fluorescence excited by different polarizations of the exciting laser beam, there is determined a localization and extent of anisotropy of absorption and/or localization and extent of fluorescence anisotropy, which is then used for determining both structural and functional information on protein properties. The present invention also discloses a suitable apparatus for acquiring structural and functional information on proteins based on polarization fluorescent microscopy wherein such apparatus comprises a modulator (P) for quick polarization modulation of an exciting beam (1) to generate a two-photon or multi-photon fluorescence and a control unit (R), wherein the operation of both the modulator (P) and the control unit (R) is synchronized by microscope (M) scanning such that information on fluorescence intensity recorded by the microscope (M) is assignable to various polarization states of the exciting beam (1) based on knowledge of a time profile of the used modulation of the exciting beam (1) polarization induced by the modulator (P).