Composition for clearing of biotissue and clarity method for biotissue using thereof

Patent metadata

Field	Value
Publication number	US-10359344-B2
Application number	US-201715622529-A
Country	US
Kind code	B2
Filing date	Jun 14, 2017
Priority date	Apr 21, 2017
Publication date	Jul 23, 2019
Grant date	Jul 23, 2019

Abstract

The present invention relates to a composition for clearing biotissue and a clarity method for biotissue using the same. The composition for clearing biotissue and the clarity method for biotissue using the same of the present invention do not need a high price electrophoresis apparatus and expensive solutions, and can not only be applied to various biotissues including brain, liver, lung, kidney, intestine, heart, muscle, and blood vessel, without damaging any of them but also prevent bubble formation, color change, and dark sediment; improve the clarity of biotissues; and help antibody-staining of the cleared tissues. Therefore, it is useful to identify the reason of various diseases through structural imaging of biotissue and to establish a treatment method.

First claim

What is claimed is:

1. A method for clarifying biotissue comprising the clearing fixed biotissue, comprising
contacting the fixed biotissue with a composition, wherein the composition comprises about 2-55% weight (w)/volume (v) % of a compound represented by formula 2 below, or the hydrate thereof

and about 10-70 w/v % of urea;
thereby clarifying the biotissue.

2. The method for clarifying biotissue according to claim 1 , wherein the fixed biotissue is fixed by one or more materials selected from the group consisting of paraformaldehyde, ethylene glycol diglycidyl ether, dipropylene glycol diglycidyl ether, 1,4-butanediol diglycidyl ether, glycerol polyglycidyl ether, glutaraldehyde and polyacrylamide.

3. The method for clarifying biotissue according to claim 1 , wherein fixed biotissue is contacted with the composition at the temperature between 4° C.-50° C. and wherein the composition comprises about 4-50 w/v % of the compound represented by formula 2 and about 20-60 w/v % urea.

4. The method for clarifying biotissue according to claim 1 , wherein the biotissue is brain, blood vessel, liver, lung, kidney, pancreas, or intestinal tissue.

5. The method for clarifying biotissue according to claim 3 , wherein the compositions comprises about 4-50 w/v % of the compound represented by formula 2 and about 35-60 w/v % of urea.

6. The method for clarifying biotissue according to claim 1 , wherein the compositions comprises about 20 w/v % of the compound represented by formula 2 and about 60 w/v % of urea.

7. The method for clarifying biotissue according to claim 1 , wherein the compositions comprises about 10 w/v % of the compound represented by formula 2 and about 65 w/v % of urea.

8. The method for clarifying biotissue according to claim 1 , wherein the compositions comprises about 40 w/v % of the compound represented by formula 2 and about 45 w/v % of urea.

Assignees

• Korea Res Inst Chemical Tech

Inventors

• Park Sunhyun
• Kim Ki-Suk

Classifications

• G01N1/30 Primary

  Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} · CPC title

• C07J41/0061

  one of the carbon atoms being part of an amide group · CPC title

• G01N1/34

  Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)} · CPC title

• G01N2001/305

  Fixative compositions · CPC title

• G01N1/28 Primary

  Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) · CPC title