US 12,111,454 B2
Laser scanning microscope and method for determining a position of a fluorophore
Arnold Giske, Sandhausen (DE); and Jonas Foelling, Freiburg (DE)
Assigned to LEICA MICROSYSTEMS CMS GMBH, Wetzlar (DE)
Appl. No. 17/779,592
Filed by Leica Microsystems CMS GmbH, Wetzlar (DE)
PCT Filed Nov. 18, 2020, PCT No. PCT/EP2020/082465
§ 371(c)(1), (2) Date May 25, 2022,
PCT Pub. No. WO2021/104955, PCT Pub. Date Jun. 3, 2021.
Claims priority of application No. 19212710 (EP), filed on Nov. 29, 2019; and application No. 19215601 (EP), filed on Dec. 12, 2019.
Prior Publication US 2023/0008453 A1, Jan. 12, 2023
Int. Cl. G02B 21/00 (2006.01); G01N 21/64 (2006.01)
CPC G02B 21/0076 (2013.01) [G01N 21/6458 (2013.01); G02B 21/0032 (2013.01); G02B 21/0048 (2013.01); G02B 21/0072 (2013.01)] 15 Claims
OG exemplary drawing
 
1. A laser scanning microscope, comprising:
a light source configured to form an illumination light beam, the illumination light beam having a transverse light intensity profile comprising an intensity minimum located at a center of the illumination light beam,
a scanning device configured to scan the illumination light beam so that the center of the illumination light beam moves along a closed trajectory in a target area of a specimen,
a detector configured to detect fluorescence light emitted by a fluorophore within the target area of the specimen, the fluorophore being excited by the illumination light beam, and
a processor configured to;
determine an intensity distribution of the fluorescence light as a function of time,
fit a periodic function to the intensity distribution of the fluorescence light, and
determine a position of the fluorophore within the target area based on at least one parameter of the fitted periodic function.