FIELD: biotechnology.
SUBSTANCE: invention relates to methods for measuring local temperature in living cells, in particular intracellular temperature. The described method consists in recording the decay kinetics of the anti-Stokes fluorescence of the TagRFP protein in cells. To measure temperature, it is proposed to use the phenomenon of conformational mobility of the chromophore of the TagRFP protein, represented by the heterogeneity of the spectral-temporal characteristics of the anti-Stokes fluorescence. This phenomenon manifests itself in a change in the characteristic decay time of the anti-Stokes fluorescence of the TagRFP protein upon heating under excitation by light quanta with a wavelength of 600 to 650 nm. The dependence of the ratio of the average characteristic fluorescence decay time to the time of the longest component on the temperature has the character of an inclined straight line.
EFFECT: possibility of functional visualization of the local temperature inside and/or on the surface of living cells and the construction of temperature maps of the entire cell or a separate area with an accuracy of 0.05°C to 0.3°C and spatial resolution up to 300 nm.
2 cl, 3 dwg, 2 ex
