US 12,243,194 B2
STED microscopy method with improved signal to noise ratio in low photon count imaging conditions
Luis Alvarez, Heidelberg (DE); and Frank Hecht, Mannheim (DE)
Assigned to LEICA MICROSYSTEMS CMS GMBH, Wetzlar (DE)
Appl. No. 17/636,035
Filed by LEICA MICROSYSTEMS CMS GMBH, Wetzlar (DE)
PCT Filed Oct. 8, 2020, PCT No. PCT/EP2020/078317
§ 371(c)(1), (2) Date Feb. 17, 2022,
PCT Pub. No. WO2021/069611, PCT Pub. Date Apr. 15, 2021.
Claims priority of application No. 19202285 (EP), filed on Oct. 9, 2019.
Prior Publication US 2022/0343467 A1, Oct. 27, 2022
Int. Cl. G06T 5/70 (2024.01); G01N 21/64 (2006.01); G06T 5/10 (2006.01); G06T 5/20 (2006.01); G06T 5/50 (2006.01)
CPC G06T 5/70 (2024.01) [G01N 21/6428 (2013.01); G01N 21/6458 (2013.01); G06T 5/10 (2013.01); G06T 5/20 (2013.01); G06T 5/50 (2013.01); G01N 2021/6439 (2013.01); G06T 2207/10056 (2013.01); G06T 2207/10064 (2013.01); G06T 2207/20064 (2013.01)] 11 Claims
OG exemplary drawing
 
1. A method for generating a result image, the method comprising the steps of:
a) acquiring a stimulated emission depletion (STED) image of a sample, the STED image comprising pixels;
b) calculating Fourier coefficients of photon arrival times for the pixels of the STED image, resulting in real coefficients representing a first image and in imaginary coefficients representing a second image;
c) deriving an intensity image from the STED image;
d) applying a spatial filter to the first image, the second image and the intensity image, resulting in a filtered first image, a filtered second image, and a filtered intensity image, respectively, wherein the spatial filter is a de-noising filter;
e) calculating an image G based on the filtered first image and the filtered intensity image by dividing the filtered first image by the filtered intensity image;
f) calculating an image S based on the filtered second image and the filtered intensity image by dividing the filtered second image by the filtered intensity image; and
g) calculating the result image based on the image G and the image S.