US 11,954,831 B2
Method of image evaluation for sim microscopy and sim microscopy method
Ingo Kleppe, Jena (DE); Yauheni Novikau, Apolda (DE); and Lutz Schaefer, Kitchener (CA)
Assigned to Carl Zeiss Microscopy GmbH, Jena (DE)
Filed by Carl Zeiss Microscopy GmbH, Jena (DE)
Filed on Sep. 7, 2021, as Appl. No. 17/467,912.
Claims priority of application No. 10 2020 123668.9 (DE), filed on Sep. 10, 2020.
Prior Publication US 2022/0092752 A1, Mar. 24, 2022
Int. Cl. G06T 5/20 (2006.01); G01B 11/25 (2006.01); G02B 21/06 (2006.01); G02B 21/36 (2006.01); G06T 5/50 (2006.01); G06T 5/73 (2024.01); G06V 10/145 (2022.01); G06V 10/88 (2022.01); G06V 20/69 (2022.01)
CPC G06T 5/20 (2013.01) [G01B 11/25 (2013.01); G02B 21/06 (2013.01); G02B 21/367 (2013.01); G06T 5/50 (2013.01); G06T 5/73 (2024.01); G06V 10/145 (2022.01); G06V 10/89 (2022.01); G06V 20/693 (2022.01); G06V 20/698 (2022.01); G06T 2207/10056 (2013.01); G06T 2207/10064 (2013.01); G06T 2207/10152 (2013.01)] 13 Claims
OG exemplary drawing
 
1. A method of image evaluation when performing SIM microscopy on a sample, comprising:
A) providing n raw images of the sample, which were each generated by illuminating the sample with an individually positioned SIM illumination pattern and imaging the sample in accordance with a point spread function,
B) providing n illumination pattern functions, which each describe one of the individually positioned SIM illumination patterns,
C) providing the point spread function, and
D) carrying out an iteration method, which comprises following iteration steps a) to e) as follows:
a) providing an estimated image of the sample,
b) generating simulated raw images, in each case by image processing of the estimated image using the point spread function and one of the n illumination pattern functions such that n simulated raw images are obtained,
c) assigning each of the n simulated raw images to that of the n provided raw images which was generated by the illumination pattern that corresponds to the illumination pattern function used to generate the simulated raw image, and calculating n correction raw images by the comparison of each provided raw image with the simulated raw image assigned thereto,
d) generating a correction image by combining image processing of the n correction raw images using the point spread function and the n illumination pattern functions, and
e) reconstructing the estimated image of the sample by means of the correction image and using the corrected estimated image of the sample as the estimated image of the sample in iteration step a) in the next run through the iteration,
carrying out a filtering step in a plurality of implementations of iteration step d), said filtering step attenuating or suppressing a signal component corresponding to an order of diffraction of the illumination pattern.