Title:
SAMPLE TREATMENT METHOD FOR IN-SITU IN VITRO CULTURE CELLS FOR OBSERVATION BY TRANSMISSION ELECTRON MICROSCOPY
Document Type and Number:
WIPO Patent Application WO/2023/029909
Kind Code:
A1
Abstract:
Disclosed is a sample treatment method for in-situ in-vitro culture cells for observation by transmission electron microscopy. The method comprises: preparing agar gel, placing the agar gel in a pore bottom of a cell culture plate, performing acidification on the agar gel, rinsing, and performing cationic activity treatment on the surface of the agar gel, and inoculating a cell to be observed onto the surface of the obtained agar gel; in subsequent sample treatment and preparation, performing osmic acid fixation and rinsing, then using a scalpel and microscopic tweezers to cut an agar gel block containing cell growth, transferring the agar gel block to another culture plate, performing subsequent dehydration and permeation, performing sample embedding in a capsule-type embedding plate, and then performing conventional slicing, dyeing, and on-machine observation. According to the described method, a change in the original growth position of a culture cell and changes in ultramicro structure, which are caused by a conventional sample treatment means and lead to errors in the experiment result, can be avoided. Meanwhile, the present invention has great significance for in-situ electron microscopy observation of culture cells and for studying the super-microstructure relationship between adjacent cells.
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Inventors:
ZHU CHANGLAI (CN)
HE XIAOQIN (CN)
LIU XIAOMAN (CN)
LIU FANG (CN)
WANG YINGJIE (CN)
HE XIAOQIN (CN)
LIU XIAOMAN (CN)
LIU FANG (CN)
WANG YINGJIE (CN)
Application Number:
PCT/CN2022/111315
Publication Date:
March 09, 2023
Filing Date:
August 10, 2022
Export Citation:
Assignee:
UNIV NANTONG (CN)
International Classes:
G01N1/28; G01N1/30; G01N1/36; G01N1/40
Foreign References:
| CN113654863A | 2021-11-16 | |||
| CN110082380A | 2019-08-02 | |||
| CN109781493A | 2019-05-21 |
Other References:
QIAN JINGJING; GUAN HUAIJIN; ZHU CHANGLAI: "Preparation method of in situ embedding electron microscope sample of cultured cells in vitro", ACTA UNIVERSITATIS MEDICINALIS NANJING (NATURAL SCIENCE) [JOURNAL OF NANJING MEDICAL UNIVERSITY (NATURAL SCIENCE)], NANJING UNIVERSITY, CN, vol. 33, no. 5, 31 May 2013 (2013-05-31), CN , pages 707 - 708, XP009544053, ISSN: 1007-4368
JIN LIANG-YUN; CHEN XIN-YUE; DU ZUN-SHU; CHANG LI-RONG; SONG YI-ZHI; LI HUI; WU YAN: "Preparation of in situ embedding electron microscopic samples of cultured neurons in vitro", DIANZI-XIANWEI-XUEBAO = JOURNAL OF CHINESE ELECTRON MICROSCOPY SOCIETY, vol. 39, no. 3, 1 January 2020 (2020-01-01), pages 334 - 336, XP009544054, ISSN: 1000-6281
MATTHEW A. GONDA, STUART A. AARONSON, NELSON ELLMORE, VICTOR H. ZEVE, KUNIO NAGASHIMA: "Ultrastructural Studies of Surface Features of Human Normal and Tumor Cells in Tissue Culture by Scanning and Transmission Electron Microscopy", JOURNAL OF THE NATIONAL CANCER INSTITUTE, OXFORD UNIVERSITY PRESS, GB, vol. 56, no. 2, 1 February 1976 (1976-02-01), GB , pages 245 - 263, XP009544055, ISSN: 0027-8874, DOI: 10.1093/jnci/56.2.245
JIN LIANG-YUN; CHEN XIN-YUE; DU ZUN-SHU; CHANG LI-RONG; SONG YI-ZHI; LI HUI; WU YAN: "Preparation of in situ embedding electron microscopic samples of cultured neurons in vitro", DIANZI-XIANWEI-XUEBAO = JOURNAL OF CHINESE ELECTRON MICROSCOPY SOCIETY, vol. 39, no. 3, 1 January 2020 (2020-01-01), pages 334 - 336, XP009544054, ISSN: 1000-6281
MATTHEW A. GONDA, STUART A. AARONSON, NELSON ELLMORE, VICTOR H. ZEVE, KUNIO NAGASHIMA: "Ultrastructural Studies of Surface Features of Human Normal and Tumor Cells in Tissue Culture by Scanning and Transmission Electron Microscopy", JOURNAL OF THE NATIONAL CANCER INSTITUTE, OXFORD UNIVERSITY PRESS, GB, vol. 56, no. 2, 1 February 1976 (1976-02-01), GB , pages 245 - 263, XP009544055, ISSN: 0027-8874, DOI: 10.1093/jnci/56.2.245
Attorney, Agent or Firm:
BEIJING FINELAND IP FIRM (CN)
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