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Method for rendering tissue transparent, reagent for rendering tissue transparent, and tissue observation method

Foreign code F170009219
File No. AF13-04WO
Posted date Sep 13, 2017
Country China
Application number 201380071465
Gazette No. 104956201
Date of filing Nov 20, 2013
Gazette Date Sep 30, 2015
International application number JP2013006811
International publication number WO2014115206
Date of international filing Nov 20, 2013
Date of international publication Jul 31, 2014
Priority data
  • P2013-012889 (Jan 28, 2013) JP
  • 2013WO-JP04369 (Jul 17, 2013) WO
  • 2013WO-JP06811 (Nov 20, 2013) WO
Title Method for rendering tissue transparent, reagent for rendering tissue transparent, and tissue observation method
Abstract (CN104956201)
A method for rendering tissue transparent, said method including a step in which tissue is immersed in a water-soluble solvent including 2,2'-thiodiethanol, and glycerol and/or a non-ionic organic iodine compound, is provided as a technique with which, using a simple operation, and without using toxic or hazardous organic solvents, various organs can be rendered sufficiently transparent without inducing changes therein.
In this method for rendering tissue transparent, it is preferable that a mixed solvent of 2,2'-thiodiethanol, glycerol, and a non-ionic organic iodine-compound aqueous solution be used for the water-soluble solvent.
Scope of claims [claim1]
1. Transparentising tissue a method, comprising the water-soluble solvent impregnated tissue of step,
Water-soluble solvent contains glycerol and a nonionic organic iodo compounds at least one of, and 2, 2 '-thiodiethanol.
[claim2]
2. Tissue transparentising method according to claim 1, wherein,
The solvent is 2, 2' -thiodiethanol, nonionic organic iodine compound mixture of glycerol and water, 2, 2 '-thiodiethanol, glycerol organoiodine compound is a nonionic iodine content of 40% aqueous solution of 10 volume ratio is -50%, 1-10 and the 20%-70%.
[claim3]
3. Tissue transparentising method according to claim 1, wherein,
The solvent is 2, 2' -nonionic organic iodine compound dithiobispyridyl mixed aqueous solution of ethanol, 2, 2 '-dithiobispyridyl organoiodine compound content is 40% ethanol and iodine aqueous solution of 20 volume ratio of nonionic-80%, 80-20%.
[claim4]
4. Tissue transparentising method according to claim 1, wherein,
The solvent is 2, 2' -glycerol mixture of thiodiethanol, 2, 2 '-volume ratio of the ethanol and glycerol dithiobispyridyl 70-95%, 5-30%.
[claim5]
5. Tissue transparentising method according to claim 1, wherein,
Aqueous sucrose solution containing solvents.
[claim6]
6. Claim 1-5 transparency method as in any one of the tissue, wherein,
[claim7]
1.4-1.7 refractive index of the solvent.
[claim8]
7. Claim 1-6 transparentising method as in any one of the tissue, wherein,
The tissue is brain, spinal cord, liver, lung, heart, blood vessel tissue of the cancer-one.
[claim9]
8. Tissue transparentising method according to claim 7, wherein
The tissue has a thickness of 1 mm or more.
[claim10]
9. Transparentising agent a tissue, comprising a water-soluble solvent, water-soluble solvent contains glycerol and at least one of a nonionic organic iodo compounds, and 2, 2' -thiodiethanol.
[claim11]
10. A method of observation, the impregnated water-soluble solvent in tissue comprising the step of enabling the transparentising, iodine containing glycerol and a nonionic water-soluble solvent having at least one compound, and 2, 2'-thiodiethanol.
[claim12]
11. Observation method according to claim 10, comprising:
The marking step, the fluorescence-labeled tissue,
Adjust step, fluorescence labeled tissue immersed in the solvent, and
The detection step, detecting a transparentized tissue fluorescence from the fluorescent label.
[claim13]
12. Observation method according to claim 10, comprising:
Adjust step, tissue immersed in the solvent,
The marking step, of tissue after transparentising fluorescent label, and
Detecting step, transparentising and the fluorescent label after detection by the fluorescence label in fluorescence emitted by the tissue.
[claim14]
13. Claim 11 or 12 as observation method, wherein,
The detection step, using a fluorescent microscope, fluorescence microscope, a confocal microscope or a multiphoton microscope fluorescence.
  • Applicant
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • ONODERA HIROSHI
IPC(International Patent Classification)
Reference ( R and D project ) CREST Creation of a Novel Technology towards Diagnosis and Treatment based on Understanding of Molecular Pathogenesis of Psychiatric and Neurological Disorders AREA
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