US 12,474,241 B2
3D image registration via dense staining and super-resolution multiplexed imaging method using the same
Jae-Byum Chang, Daejeon (KR); In Cho, Daejeon (KR); Chan E Park, Daejeon (KR); Jueun Sim, Daejeon (KR); and Young-Gyu Yoon, Daejeon (KR)
Assigned to Korea Advanced Institute Of Science And Technology, Daejeon (KR)
Filed by Korea Advanced Institute Of Science And Technology, Daejeon (KR)
Filed on Feb. 18, 2022, as Appl. No. 17/675,808.
Claims priority of application No. 10-2021-0024480 (KR), filed on Feb. 24, 2021; application No. 10-2021-0138718 (KR), filed on Oct. 18, 2021; application No. 10-2022-0009372 (KR), filed on Jan. 21, 2022; application No. 10-2022-0010077 (KR), filed on Jan. 24, 2022; and application No. 10-2022-0010078 (KR), filed on Jan. 24, 2022.
Prior Publication US 2022/0268672 A1, Aug. 25, 2022
Int. Cl. G01N 1/30 (2006.01); G06T 7/33 (2017.01)
CPC G01N 1/30 (2013.01) [G06T 7/337 (2017.01); G01N 2001/302 (2013.01); G06T 2200/04 (2013.01); G06T 2207/10056 (2013.01); G06T 2207/30204 (2013.01)] 7 Claims
OG exemplary drawing
 
1. A three-dimensional image registration method comprising:
placing at least one fiducial marker in an internal structure of a biological sample via dense staining;
obtaining a plurality of images by repeatedly imaging the biological sample while replacing a fluorescent label attached to at least one target molecule within the biological sample where the fiducial marker is placed; and
registering the images using the fiducial marker as a reference,
wherein, in the obtaining of images, the biological sample is repeatedly imaged in a plurality of imaging rounds to obtain each of the images in each of the imaging rounds,
wherein the biological sample is produced by applying an expandable polymer material to a biological specimen and expanding the expandable polymer material,
wherein the three-dimensional image registration method further comprises
in the obtaining of images, extracting a vector map in which both physical deformities in the biological sample caused by repeatedly mounting the biological sample on a microscope, and non-linear deformities within the expandable polymer material caused by repeated replacement of the fluorescent label, are mapped, and
wherein the registering of the images comprises obtaining a final image of the target molecule by overlaying the images using the fiducial marker and the vector map as a reference.