Title:
CLEARING AND EXPANSION METHOD AND IMAGING METHOD FOR BIOLOGICAL TISSUE
Document Type and Number:
WIPO Patent Application WO/2023/029425
Kind Code:
A1
Abstract:
Provided are a clearing and expansion method and an imaging method for biological tissue. The clearing and expansion method for biological tissue comprises: (1) degreasing a fixed biological tissue sample; (2) soaking the degreased biological tissue sample in a solution of gel monomer molecules, such that the monomer molecules permeate into the degreased biological tissue sample; (3) inducing a polymerization reaction of the monomer molecules permeating into the biological tissue sample so as to form a polymer gel; and (4) placing the biological tissue sample which has been subjected to a gelation treatment into water for expansion. The clearing and expansion method for biological tissue avoids the sample treatment steps of heating and enzymatic digestion that cause fluorescent quenching and the destruction of biological tissue, and thus has the advantages of high retention of endogenous fluorescent proteins, high mechanical strength of expanded biological tissue, a short sample treatment time, etc. Moreover, the advantage of an adjustable expansion ratio is further achieved by means of changing the components of a monomer reagent.
Inventors:
GAO LIANG (CN)
CHEN YANLU (CN)
LU JING (CN)
FENG RUILI (CN)
CHEN YANLU (CN)
LU JING (CN)
FENG RUILI (CN)
Application Number:
PCT/CN2022/079854
Publication Date:
March 09, 2023
Filing Date:
March 09, 2022
Export Citation:
Assignee:
UNIV WESTLAKE (CN)
International Classes:
G01N1/28; G01N1/36; G01N1/44; G01N21/64
Domestic Patent References:
| WO2017188264A1 | 2017-11-02 |
Foreign References:
| US20200217761A1 | 2020-07-09 | |||
| CN106872252A | 2017-06-20 | |||
| CN111492223A | 2020-08-04 | |||
| CN106769350A | 2017-05-31 | |||
| US20190145868A1 | 2019-05-16 | |||
| KR101888782B1 | 2018-08-14 |
Other References:
MATSUMOTO KATSUHIKO; MITANI TOMOKI T.; HORIGUCHI SHUHEI A.; KANESHIRO JUNICHI; MURAKAMI TATSUYA C.; MANO TOMOYUKI; FUJISHIMA HIROS: "Advanced CUBIC tissue clearing for whole-organ cell profiling", NATURE PROTOCOLS, NATURE PUBLISHING GROUP, GB, vol. 14, no. 12, 20 November 2019 (2019-11-20), GB , pages 3506 - 3537, XP036929691, ISSN: 1754-2189, DOI: 10.1038/s41596-019-0240-9
WANG, PEIXIN; ZHANG, DAN; SHANG, AIJIA; HOU, BING: "Tissue Optical Clearing Technique", CHINESE JOURNAL OF NEUROANATOMY, SPRINGER-VERLAG, NEW YORK, vol. 32, no. 1, 31 December 2016 (2016-12-31), New York , pages 124 - 128, XP009544234, ISSN: 1000-7547, DOI: 10.16557/j.cnki.1000-7547.2016.01.023
YU, TINGTING ET AL.: "Review of Tissue Optical Clearing Methods for Imaging Whole Organs", CHINESE JOURNAL OF LASERS, SHANGHAI : SHANGHAI KEXUE JISHU CHUBANSHE, 1983-1992, CN, vol. 47, no. 2, 29 February 2020 (2020-02-29), CN , pages 103 - 119, XP009544233, ISSN: 0258-7025
MURAKAMI, T. C. ET AL.: "A three-dimensional single-cell-resolution whole-brain atlas using CUBIC-X expansion microscopy and tissue clearing", NATURE NEUROSCIENCE, vol. 21, 30 April 2018 (2018-04-30), pages 625 - 637, XP036468142, DOI: 10.1038/s41593-018-0109-1
TAINAKA KAZUKI, MURAKAMI TATSUYA C., SUSAKI ETSUO A., SHIMIZU CHIKA, SAITO RIE, TAKAHASHI KEI, HAYASHI-TAKAGI AKIKO, SEKIYA HIROSH: "Chemical Landscape for Tissue Clearing Based on Hydrophilic Reagents", CELL REPORTS, ELSEVIER INC, US, vol. 24, no. 8, 1 August 2018 (2018-08-01), US , pages 2196 - 2210.e9, XP093042954, ISSN: 2211-1247, DOI: 10.1016/j.celrep.2018.07.056
WANG, PEIXIN; ZHANG, DAN; SHANG, AIJIA; HOU, BING: "Tissue Optical Clearing Technique", CHINESE JOURNAL OF NEUROANATOMY, SPRINGER-VERLAG, NEW YORK, vol. 32, no. 1, 31 December 2016 (2016-12-31), New York , pages 124 - 128, XP009544234, ISSN: 1000-7547, DOI: 10.16557/j.cnki.1000-7547.2016.01.023
YU, TINGTING ET AL.: "Review of Tissue Optical Clearing Methods for Imaging Whole Organs", CHINESE JOURNAL OF LASERS, SHANGHAI : SHANGHAI KEXUE JISHU CHUBANSHE, 1983-1992, CN, vol. 47, no. 2, 29 February 2020 (2020-02-29), CN , pages 103 - 119, XP009544233, ISSN: 0258-7025
MURAKAMI, T. C. ET AL.: "A three-dimensional single-cell-resolution whole-brain atlas using CUBIC-X expansion microscopy and tissue clearing", NATURE NEUROSCIENCE, vol. 21, 30 April 2018 (2018-04-30), pages 625 - 637, XP036468142, DOI: 10.1038/s41593-018-0109-1
TAINAKA KAZUKI, MURAKAMI TATSUYA C., SUSAKI ETSUO A., SHIMIZU CHIKA, SAITO RIE, TAKAHASHI KEI, HAYASHI-TAKAGI AKIKO, SEKIYA HIROSH: "Chemical Landscape for Tissue Clearing Based on Hydrophilic Reagents", CELL REPORTS, ELSEVIER INC, US, vol. 24, no. 8, 1 August 2018 (2018-08-01), US , pages 2196 - 2210.e9, XP093042954, ISSN: 2211-1247, DOI: 10.1016/j.celrep.2018.07.056
Attorney, Agent or Firm:
KINGSOUND & PARTNERS (CN)
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