IP Library Granted Patent US 12,560,796
Granted Patent B2
US 12,560,796 · App. 18/608,479 · Granted Feb 24, 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 12,560,796
App. No.
18/608,479
Granted
Feb 24, 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 (20)

1 . A method, comprising the steps of:

acquiring a first image of a first field of view of a first subset of a biological sample;

processing the first image automatically based on a predefined criterion to determine first coordinate information of a first region of interest of the first image;

illuminating the first field of view with a first light pattern corresponding to the first coordinate information of the first region of interest of the first image;

acquiring a second image of a second field of view of a second subset of the biological sample;

processing the second image automatically based on the predefined criterion to determine second coordinate information of a second region of interest of the second image;

illuminating the second field of view with a second light pattern different from the first light pattern and corresponding to the second coordinate information of the second region of interest of the second image.

2 . The method according to claim 1 , further comprising controlling a camera to acquire the first and the second images.

3 . The method according to claim 1 , further comprising controlling an imaging light source to provide imaging light to the biological sample when acquiring the first and second images.

4 . The method according to claim 1 , further comprising controlling an illuminating light source to provide illuminating light through an illuminating light path to illuminate the first and second fields of view of the biological sample to induce a photochemical reaction in the first and second regions of interest of the biological sample.

5 . The method according to claim 4 , further comprising controlling a pattern illumination device comprising a pair of scanning mirrors to direct the illuminating light to at least one targeted point within the first and second fields of view.

6 . The method according to claim 1 , wherein the first and second images are acquired through a microscope.

7 . The method according to claim 6 , further comprising loading the biological sample on a stage of the microscope.

8 . The method according to claim 1 , wherein a first processing module is used to perform the acquiring and processing steps.

9 . The method according to claim 8 , wherein a second processing module is used to perform the illuminating steps.

10 . The method according to claim 1 , further comprising moving the stage from the first field of view to the second field of view.

11 . The method according to claim 4 , wherein the photochemical reaction results in plurality of biotinylated amino acids in the first and second regions of interest of the biological sample.

12 . The method according to claim 11 , further comprising purifying the biotinylated amino acids to perform mass spectrometry for proteomic analysis.

13 . The method according to claim 1 , further comprising incubating the biological sample with a photosensitizer, a chemical agent and a tagging reagent before acquiring step, wherein the photosensitizer, the chemical agent and the tagging reagent are a single molecule with all three functions.

14 . The method according to claim 13 , wherein the chemical agent is conjugated with tagging reagent, and the photosensitizer and the chemical agent are separate molecules.

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 20240219704A1 · Jul 4, 2024
References Cited (123)
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 · 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 · Lyer 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 20240219705A1 · Liao et al. · 2024 [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]
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]
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.har… [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. 18/997,408 entitled “Training Al 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]