Patent ID: 6424421
Filing Date: 2002-07-23
Classification: B82Y,G02B

Abstract:
A method for applying far field microscopy and/or flow fluorometry to geometric distance measurements between object structures, wherein the distances can be smaller than the half-intensity width of the principle maximum of the point spread function, comprising the steps of:marking measurement structures to be investigated or localised with fluorescent dyes before, during or after preparation of an object on or in an object carrier, wherein at least the measurement structures to be localised whose distance is smaller than a half-intensity width of a principle maximum of a point spread function are marked with fluorescent dyes with different spectral signatures, marking calibration targets with a defined size and array with the same fluorescent dyes, preparing the fluorescent calibration targets either together with the measurement structures in or on an object carrier or separately in or on an object carrier, investigating the measurement structures and calibration targets under corresponding conditions, measuring two calibration targets with different spectral signatures to obtain actual measured values, dependent imaging and localisation behaviour of the optical system used, the measurement determining a difference between the actual measured values and predetermined known real distance values to obtain a correction value using the correction value to correct a shift brought about by the optical system in the detection of different emission loci.