Method and device for recognizing dark states during the spectroscopic or microscopic examination of fluorescent specimens
AssigneeLeica Microsystems CMS GmbH
InventorsJuergen Riedmann, Werner Knebel, Lioba Kuschel
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Abstract
A method for recognizing dark states during the spectroscopic or microscopic examination of fluorescent specimens (1), in particular using fluorescent proteins, the excitation/illumination volume, thus the intensity distribution of the excitation light, being varied over the course of at least two mutually independent measurements, a determination being made as to whether the observed time constants change and, in the case of unchanged time constants, the existence of a dark state being inferred.
Claims28
A method for recognizing dark states during the spectroscopic or microscopic examination of fluorescent specimens, the method comprising:
- varying an intensity distribution of excitation light by varying an excitation/illumination volume over a plurality of mutually independent measurements;
- and
- determining whether respective time constants change between the measurements, and inferring an existence of a dark state where the respective time constants are unchanged between the measurements.
A device for spectroscopically or microscopically examining fluorescent specimens, the device including:
- a light source configured to produce excitation light;
- a detector;
- an illumination beam path extending between the light source and a specimen;
- a detection beam path extending between a specimen and the detector;
- an excitation/illumination volume variation device configured to vary an intensity distribution of the excitation light by varying an excitation/illumination volume over a course of a plurality of mutually independent measurements;
- and
- a time constant measurement device configured to measure respective time constants so as to enable a determination of whether the measured respective time constants change between the measurements and an inferring of an existence of a dark state where the measured respective time constants are unchanged between the measurements.
Dependencies are inferred from the claim text.