IP Library Granted Patent US 12,535,669
Granted Patent B2
US 12,535,669 · App. 18/608,490 · Granted Jan 27, 2026

Microscope-based system and method for image-guided microscopic illumination

Inventors: Jung-Chi Liao (Taipei, TW); Yi-De Chen (Taipei, TW); Chih-Wei Chang (Taipei, TW); Weng Man Chong (Taipei, TW)
Assignee: Academia Sinica
G02B21/365G02B21/06G02B21/26G06V10/141G06V10/25G06V20/693H04N23/56H04N23/74
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Quick Facts
Patent No.
US 12,535,669
App. No.
18/608,490
Granted
Jan 27, 2026
Kind
B2
Abstract

A system and method for image-guided microscopic illumination are provided. A processing module controls an imaging assembly such that a camera acquires an image or images of a sample in multiple fields of view, and the image or images are automatically transmitted to a processing module and processed by the first processing module automatically in real-time based on a predefined criterion so as to determine coordinate information of an interested region in each field of view. The processing module also controls an illuminating assembly to illuminate the interested region of the sample according to the received coordinate information regarding to the interested region, with the illumination patterns changing among the fields of view.

Claims (19)

1 . A system for proximity photolabeling of target proteins in a biological sample, comprising:

a microscope comprising a biological sample loaded on a stage;

an imaging assembly comprising a camera and an imaging light source, wherein the imaging assembly is configured to acquire at least one image of a plurality of fields of view of the biological sample;

an illumination assembly comprising a photosensitizing light source and a pattern illumination device, wherein the photosensitizing light source is configured to induce a photochemical reaction in the biological sample, and wherein the pattern illumination device is configured to control the laser to illuminate a plurality of patterns in the plurality of fields of view of the biological sample;

at least one processor operatively coupled to the imaging assembly, the illumination assembly, and the stage, the at least one processor being configured to:

control the imaging assembly to acquire a first image in a first field of view of the biological sample;

process the first image to determine a first region of interest of the first field of view of the biological sample;

control the illumination assembly to illuminate the first field of view with a first light pattern that corresponds to the first region of interest; and

after the first region of interest of the first field of view has been illuminated, control the stage to a subsequent field of view of the biological sample.

2 . The system according to claim 1 , wherein the processor is further configured to:

control the imaging assembly to acquire a second image in the subsequent field of view;

process the second image to determine a second region of interest of the subsequent field of view;

control the illumination assembly to illuminate the second field of view with a second light pattern that corresponds to the second region of interest and is different than the first light pattern.

3 . The system according to claim 1 , wherein the pattern illumination device comprises a pair of scanning mirrors, a digital micromirror device, or a spatial light modulator, which is disposed between the photosensitizing light source and the microscope along an illuminating light path.

4 . The system according to claim 1 , wherein the photosensitizing light source is configured to produce free radical release of a photosensitizer resulting in biotinylation of amino acids in the first region of interest of the biological sample.

5 . The system according to claim 4 , wherein the system further comprises a mass spectrometer configured to perform proteomic analysis of the biotinylated biological sample.

6 . The system according to claim 1 , wherein the photosensitizing light source comprises a femtosecond laser to generate a two-photon effect for high axial illumination precision.

7 . The system according to claim 1 , wherein the processor is configured to determine the first region of interest of the biological sample based on a predefined criterion to obtain first coordinate information corresponding to the first region of interest.

8 . The system according to claim 7 , wherein the predefined criterion comprises a morphology, intensity, contrast, or specific features of the first image.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 2, 2024
From: LIAO, JUNG-CHI; CHEN, YI-DE; CHANG, CHIH-WEI; CHONG, WENG MAN
To: ACADEMIA SINICA
Reel/Frame 066983/0661 →
Continuity (5)
Continuation 18464125 · Sep 8, 2023
Continuation 17592211 · Feb 3, 2022
Continuation 16013663 · Jun 20, 2018
Provisional Application 62522265 · Jun 20, 2017
Related Publication 20240219705A1 · Jul 4, 2024
References Cited (124)
US 5646413A · Nishi · 1997 [cited by applicant]
US 6522774B1 · Bacus et al. · 2003 [cited by applicant]
US 6614031B2 · Engelhardt et al. · 2003 [cited by applicant]
US 6697526B1 · Abe · 2004 [cited by applicant]
US 6947583B2 · Ellis et al. · 2005 [cited by applicant]
US 6956647B2 · Foster et al. · 2005 [cited by applicant]
US 7133543B2 · Verwoerd et al. · 2006 [cited by applicant]
US 7460732B2 · Recht · 2008 [cited by applicant]
US 7639357B2 · Okugawa · 2009 [cited by applicant]
US 7817273B2 · Bahatt et al. · 2010 [cited by applicant]
US 8187866B2 · Duer · 2012 [cited by applicant]
US 8288157B2 · Duer · 2012 [cited by applicant]
US 8923568B2 · Olson et al. · 2014 [cited by applicant]
US 9019598B2 · Tsurumune · 2015 [cited by applicant]
US 9030547B2 · Vizi et al. · 2015 [cited by applicant]
US 9125562B2 · Spencer et al. · 2015 [cited by applicant]
US 9217694B2 · Sieckmann et al. · 2015 [cited by applicant]
US 9528939B2 · Duer · 2016 [cited by applicant]
US 9557156B2 · Kankaria · 2017 [cited by applicant]
US 9581801B2 · Takamizawa · 2017 [cited by applicant]
US 9696535B2 · Prakash et al. · 2017 [cited by applicant]
US 9788790B2 · Black et al. · 2017 [cited by applicant]
US 9810892B2 · Prakash et al. · 2017 [cited by applicant]
US 9939381B1 · Kimmel et al. · 2018 [cited by applicant]
US 10517482B2 · Sato et al. · 2019 [cited by applicant]
US 10729326B2 · Spencer et al. · 2020 [cited by applicant]
US 10746980B2 · Kenny et al. · 2020 [cited by applicant]
US 10932670B2 · Smith et al. · 2021 [cited by applicant]
US 11265449B2 · Liao et al. · 2022 [cited by applicant]
US 11408821B2 · Boamfa et al. · 2022 [cited by applicant]
US 11513328B2 · Pannhoff et al. · 2022 [cited by applicant]
US 11789251B2 · Liao et al. · 2023 [cited by applicant]
US 20020008904A1 · Engelhardt · 2002 [cited by applicant]
US 20030189676A1 · Kato et al. · 2003 [cited by applicant]
US 20030231791A1 · Torre-Bueno et al. · 2003 [cited by applicant]
US 20040105000A1 · Yuri · 2004 [cited by applicant]
US 20050072920A1 · Inada · 2005 [cited by applicant]
US 20050082494A1 · Motomura · 2005 [cited by applicant]
US 20050089208A1 · Dong et al. · 2005 [cited by applicant]
US 20050179899A1 · Palti Wasserman et al. · 2005 [cited by applicant]
US 20060028717A1 · Dunn · 2006 [cited by applicant]
US 20060226375A1 · Maruo · 2006 [cited by applicant]
US 20070152130A1 · Fomitchov · 2007 [cited by applicant]
US 20080206752A1 · Balakirev et al. · 2008 [cited by applicant]
US 20090195646A1 · Ganser et al. · 2009 [cited by applicant]
US 20090316259A1 · Kenny et al. · 2009 [cited by applicant]
US 20100079857A1 · Sasaki et al. · 2010 [cited by applicant]
US 20100278399A1 · Bednarkiewicz et al. · 2010 [cited by applicant]
US 20110050745A1 · Jung et al. · 2011 [cited by applicant]
US 20120242817A1 · Pan · 2012 [cited by applicant]
US 20130294645A1 · Sibarita · 2013 [cited by applicant]
US 20140022373A1 · Kanarowski et al. · 2014 [cited by applicant]
US 20140152794A1 · Takahashi · 2014 [cited by applicant]
US 20140293407A1 · Amano et al. · 2014 [cited by applicant]
US 20150168702A1 · Harris · 2015 [cited by applicant]
US 20150185456A1 · Kishima · 2015 [cited by applicant]
US 20150258352A1 · Lin et al. · 2015 [cited by applicant]
US 20160124208A1 · Best et al. · 2016 [cited by applicant]
US 20160302740A1 · Iyer et al. · 2016 [cited by applicant]
US 20170059407A1 · Shiraiwa · 2017 [cited by applicant]
US 20170090176A1 · Pospiech et al. · 2017 [cited by applicant]
US 20170154236A1 · Masuura et al. · 2017 [cited by applicant]
US 20170212342A1 · Rozsa et al. · 2017 [cited by applicant]
US 20170248837A1 · Sato et al. · 2017 [cited by applicant]
US 20170299784A1 · Mikkelsen et al. · 2017 [cited by applicant]
US 20180136451A1 · Soenksen · 2018 [cited by applicant]
US 20180177401A1 · Yang · 2018 [cited by examiner]
US 20180180642A1 · Shetty et al. · 2018 [cited by applicant]
US 20180180885A1 · Holland et al. · 2018 [cited by applicant]
US 20180203221A1 · Dai et al. · 2018 [cited by applicant]
US 20180210183A1 · Nishikawa · 2018 [cited by applicant]
US 20180284419A1 · Peschka et al. · 2018 [cited by applicant]
US 20180348500A1 · Naaman et al. · 2018 [cited by applicant]
US 20190324240A1 · Shroff et al. · 2019 [cited by applicant]
US 20190339456A1 · Ruggles · 2019 [cited by applicant]
US 20190391078A1 · Cohen et al. · 2019 [cited by applicant]
US 20210224954A1 · Guo et al. · 2021 [cited by applicant]
US 20230418041A1 · Liao et al. · 2023 [cited by applicant]
US 20240411122A1 · Liao et al. · 2024 [cited by applicant]
US 20250028162A1 · Liao et al. · 2025 [cited by applicant]
US 20250028163A1 · Liao et al. · 2025 [cited by applicant]
US 20250035910A1 · Liao et al. · 2025 [cited by applicant]
US 20250052993A1 · Liao et al. · 2025 [cited by applicant]
TW 201013817A · 2010 [cited by applicant]
WO WO2016133787A1 · 2016 [cited by applicant]
Alqattan et al.; Direct laser writing of nanophotonic structures on contact lenses; ACS nano; 12(6); pp. 5130-5140; Apr. 2018. [cited by applicant]
Aznar et al.; Gated materials for on-command release of guest molecules; Chemical reviews; 116(2); pp. 561-718; Jan. 27, 2016. [cited by applicant]
Bar et al.; Biotinylation by antibody recognition—a method for proximity labeling; Nature Methods; 15(2); pp. 127-133; (author Manuscript); 19 pages; Feb. 2018. [cited by applicant]
Brieke et al.; Light?controlled tools; Angewandte Chemie International Edition; 51(34); pp. 8446-8476; Aug. 20, 2012. [cited by applicant]
Chen et al.; Lattice light-sheet microscopy: imaging molecules to embryos at high spatiotemporal resolution; Science; 346(6208); 1257998; doi:10.1126/scince.1257998; (Author Manuscript); 28 pages; Oct. 2014. [cited by applicant]
Ezzoukhry et al.; Combining laser capture microdissection and proteomics reveals an active translation machinery controlling formation; Nature Communications; 9(1); doi:10.1038/s41467-018-04461-9; 11 pages; May 2018. [cited by applicant]
Hadley et al.; Determining composition of micron-scale protein deposits in neurodegenerative disease by spatially targeted optical microproteomics; eLife; 21 pages. doi: 10.7554/eLife.09579; Sep. 2015. [cited by applicant]
Hirsch et al.; Easily reversible desthiobiotin binding to streptavidin, avidin, and other bitin-binding proteins: uses for protein labeling, detection, and isolation; Analytical Biochemistry; 308(2); pp. 343-357; Sep. 2… [cited by applicant]
Hung et al.; Protein localization in disease and therapy; Journal of Cell Science; 124(20); pp. 3381-3392; Oct. 2011. [cited by applicant]
Hung et al.; Spatially resolved proteomic mapping in living cells with the engineered peroxidase APEX2; Nature Protocols; 11(3); pp. 456-475; (Author Manuscript); 39 pages; Mar. 2016. [cited by applicant]
Itzhak et al.; A mass spectrometry-based approach for mapping protein neurons; Cell Reports; 20(11); pp. 2706-2718; Sep. 2017. [cited by applicant]
Jain et al.; ATPase-modulated stress granules contain a diverse proteome and substructure; Cell; 164(3); pp. 487-498; Jan. 2016. [cited by applicant]
Kamburov et al.; The consensusPathDB interaction database: 2013 update; Nucleic Acids Research; 41(D1); pp. D793-D800; Jan. 2013. [cited by applicant]
Kepiro et al.; Molecular tattoo: subcellular confinement of drug effects; Chemistry and Biology; 22(4); pp. 548-558; Apr. 2015. [cited by applicant]
Lam et al.; Directed evolution of APEX2 for electron microscopy and proximity labeling; Nature Methods; 12(1); pp. 51-54; (author Manuscript); 15 pages; Jan. 2015. [cited by applicant]
Liao et al.; A pioneering high content cell image-registered protein labeling system; 2017 Bio Taiwan Exhibition and Conference; 2 pages; Jun. 29-Jul. 2, 2017. [cited by applicant]
Liao et al.; High content cell image-registered protein labeling system; 2017 Bio Taiwan Exhibition and Conference; 14 pages; Jun. 29-Jul. 2, 2017. [cited by applicant]
Liu et al; Supplementary materials: super-resolution labeling with action-paint; Nature Chemistry; Supplementary Information; doi.org/10.1038/s41557-019-0325-7; 43 pages retrieved from the internet (https://yin.hms.harv… [cited by applicant]
Prier et. al.; Visible light photoredox cataysis with transition metal complexes: applications in organic synthesis; Chemical reviews; 113(7); pp. 5322-5363; (author Manuscript) 141 pages; Jul. 2013. [cited by applicant]
Rhee et al.; Proteomic mapping of mitochondria in living cells via spatially restricted enzymatic tagging; Science; 339(6125); pp. 1328-1331; (Author Manuscript); 8 pages; Mar. 2013. [cited by applicant]
Rost et al.; Multiplexed ion beam imaging analysis for quanitation of protein expression in cancer tissue sections; Laboratory Investigation; 97(8); pp. 992-1003; Aug. 2017. [cited by applicant]
Roux et al.; A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells; Journal of Cell Biology; 196(6); pp. 801-810; Mar. 2012. [cited by applicant]
Roux et al.; BioID: A screen for protein-protein interactions: Current Protocols in protein Science; 91(1); pp. 19-23; (Author Manuscript); 20 pages; Jan. 2018. [cited by applicant]
Thermofisher Scientific: Sulfo-SBED biotin label transfer reagent; (Product Discription); retrieved from the internet (https://www.thermofisher.com/order/catalog/product/33033#/33033); 5 pages; on May 5, 2021. [cited by applicant]
Thul et al.; A subcellular map of the human proteome; Science; 356(6340) eaal3321; 14 pages; May 2017. [cited by applicant]
Zipfel et al.; Nonlinear magic: multiphoton microscopy in the biosciences; Nature Biotechnolgy; 21(11); pp. 1369-1377; Nov. 2003. [cited by applicant]
Liao et al.; U.S. Appl. No. 18/608,452 entitled “Microscope-based system and method for image-guided microscopic illumination,” filed Mar. 18, 2024. [cited by applicant]
Liao et al.; U.S. Appl. No. 18/608,479 entitled “Microscope-based system and method for image-guided microscopic illumination,” filed Mar. 18, 2024. [cited by applicant]
Chang et al.; U.S. Appl. No. 18/611,306 entitled “Photoreactive and cleavable probes for tagging biomolecules,” filed Mar. 20, 2024. [cited by applicant]
Ronneberger et al.; U-net: Convolutional networks for biomedical image segmentation. InMedical image computing and computer-assisted interventionRMICCAI 2015; 18th international conference; Springer international publis… [cited by applicant]
Stegmaier et al.; Real-time three-dimensional cell segmentation in large-scale microscopy data of developing embryos. Developmental Cell; 36(2); pp. 225-240; Jan. 25, 2016. [cited by applicant]
Wikipedia; Biotinylation; 4 pages; retrieved from the internet (https://en.wikipedia.org/w/index.php?title=Biotinylation&oldid=841539236) on May 16, 2018. [cited by applicant]
Huang et al.; U.S. Appl. No. 18/748,710 entitled “Adjustable Support and Optical Module,” filed Jun. 20, 2024. [cited by applicant]
Liao et al.; U.S. Appl. No. 18/864,988 entitled “Method of calibrating a microscope system,” filed Nov. 12, 2024. [cited by applicant]
Liao et al.; U.S. Appl. No. 18/864,997 entitled “Method of calibrating a microscope system,” filed Nov. 12, 2024. [cited by applicant]
Chen et al.; U.S. Appl. No. 18/877,609 entitled “Microscope-based system and method using a UV-transmission mirror,” filed Dec. 20, 2024. [cited by applicant]
Liao et al.; U.S. Appl. No. 18/997,408 entitled “Training AI model for a microscope-based pattern photoillumination system,” filed Jan. 21, 2025. [cited by applicant]
Liao et al.; U.S. Appl. No. 18/997,412 entitled “Method of standardizing image pixel values in a microscope-based system,” filed Jan. 21, 2025. [cited by applicant]
Liao et al.; U.S. Appl. No. 19/110,325 entitled “Microscope-based system and method of determining beam processing path,” filed Mar. 10, 2025. [cited by applicant]