[특허]Apparatus and methods for three-dimensional tissue measurements based on controlled media flow

Apparatus and methods for three-dimensional tissue measurements based on controlled media flow 원문보기

IPC분류정보
국가/구분	United States(US) Patent 등록
국제특허분류(IPC7판)	G01N-033/00 G01N-033/50 B01L-003/00 C12M-001/32 C12M-001/12 C12M-001/00 C12M-001/34 C12M-001/02 G01N-021/64
출원번호	US-0079022 (2013-11-13)
등록번호	US-9494577 (2016-11-15)
발명자 / 주소	McGarr, Paul Neilson, Andy C. Teich, Jay S.
출원인 / 주소	Seahorse Biosciences
인용정보	피인용 횟수 : 0 인용 특허 : 127

초록

An apparatus including a plurality of wells for conducting analysis of three-dimensional cell samples (e.g., tissue samples) and methods for experimenting with a three-dimensional sample. A removable insert for use with the apparatus enables plunger-driven perfusion of the three-dimensional sample.

대표청구항 ▼

1. An apparatus for conducting analysis of a three-dimensional cell sample, the apparatus comprising: a plurality of wells for holding respective samples and sample media, at least one of the wells comprising a sample nesting site disposed in a bottom surface of the well,disposed thereabove a bore dimensioned to interfit with a plunger which moves vertically down within and relative to the bore and within the sample media disposed in the well, anda media channel defined by the well, a removable well insert, or both the well and the insert, in fluid communication with the sample nesting site, which permits flow of the sample media therethrough impelled by the plunger and permits exposure of the sample to flowing media as the plunger moves. 2. The apparatus of claim 1 wherein the plunger moves relative to the nesting site to induce perfusion of media about the sample. 3. The apparatus of claim 1 wherein said media channel comprises a fluid path which returns media perfused about the sample back to media disposed in the well. 4. The apparatus of claim 1 wherein at least said bore is defined by the removable well insert. 5. The apparatus of claim 1 wherein the well comprises a sump in fluid communication with the media channel. 6. The apparatus of claim 1 wherein said sample nesting site is disposed within a sump for collection of media and in fluid communication with said media channel. 7. The apparatus of claim 1 wherein said media channel comprises a fluid path defining a closed loop beneath the surface of media in the wells to permit media perfusive flow about said sample on both upward and downward movement of a plunger within said bore. 8. The apparatus of claim 1 further comprising a sensor for detecting the concentration of a solute in media disposed about said sample. 9. The apparatus of claim 1 comprising a plurality of plungers adapted for reciprocating movement within the bores of respective wells. 10. The apparatus of claim 1 wherein the plurality of wells defines a multiwell plate comprising 24 or 96 wells. 11. The apparatus of claim 1 further comprising a source of oxygen, carbon dioxide, and/or a biologically inert gas in fluid communication with media in a well or a headspace above the surface of media in the wells for controlling the composition of gas in the headspace or in the media. 12. The apparatus of claim 1 further comprising a source of a solution of a biologically active substance in fluid communication with media in wells for exposing a sample to said substance. 13. The apparatus of claim 1 further comprising a three-dimensional cell growth scaffold disposed on said sample nesting site. 14. The apparatus of claim 1 further comprising a check valve disposed in said media channel to inhibit media perfusion from the channel back to the sample. 15. An apparatus for conducting analysis of three-dimensional cell samples, the apparatus comprising: a well for holding a sample and sample media comprising a sample nesting site disposed in a bottom surface of the well,disposed above said nesting site, a bore dimensioned to interfit with a plunger which moves vertically down within the bore and within media disposed in said well,a media channel in fluid communication with said sample nesting site which permits media perfusion about said sample, wherein the media channel is defined by the well, a removable well insert, or both the well and the insert, anda plunger adapted for reciprocal movement in said bore thereby to impel media perfusion about said sample,wherein the media channel permits flow of the sample media disposed in the well therethrough impelled by the plunger and permits exposure of the sample to flowing media as the plunger moves. 16. The apparatus of claim 15 further comprising a sensor for detecting the concentration of a dissolved media component in media about the sample nesting site. 17. A method of impelling media flow about a three-dimensional cell culture sample, so as to maintain viability of the sample and exercise control over its microenvironment, the method comprising the steps of: providing a well including a sample nesting site disposed in a bottom surface of the well, a bore dimensioned to interfit with a plunger disposed above the sample nesting site, a media channel in fluid communication with said sample nesting site, wherein the media channel is defined by the well, a removable well insert, or both the well and the insert, and a plunger adapted for reciprocal movement in said bore;placing a sample on the sample nesting site in medium within the well, andmoving said plunger within the bore to impel media flow about the sample and through said channel thereby to perfuse the sample with media,wherein the media channel permits flow of the sample media disposed in the well therethrough impelled by the plunger and permits exposure of the sample to flowing media as the plunger moves. 18. The method of claim 17 comprising the additional step of adding a gas to the media or to headspace in the well above the media to modify the microenvironment about the sample by altering dissolved gas composition. 19. The method of claim 17 comprising the additional step of adding a solution of a biologically active substance to the media to modify the microenvironment about the sample by exposing the sample to the biologically active substance. 20. The method of claim 17 comprising the additional step of measuring the concentration of one or more solutes in media about said sample. 21. The method of claim 17 comprising the additional step of making a plurality of measurements separated in time of the concentration of one or more solutes in media about said sample. 22. The method of claim 17 comprising the additional step of adding at least one of a metered amount of one or more gases and one or more solutes to media in said well thereby to set the microenvironment in the medium about said sample to a predetermined point. 23. The method of claim 17 wherein the sample is a tumor sample, the method comprising the additional step of adding a metered amount of one or more gases and/or one or more solutes to media in said well thereby to set the microenvironment in the medium about said sample to a predetermined point simulating the microenvironment of the tumor sample in vivo. 24. The method of claim 17 comprising multiplexing said method by the steps of: providing a plurality of said wells including sample nesting sites, bores dimensioned to interfit with plungers disposed above the sample nesting sites, media channels in fluid communication with said sample nesting site, and plungers adapted for reciprocal movement in said bore;placing a sample on the sample nesting sites in media within the plurality of wells,moving the plurality of said plungers within the bores to impel media flow about the samples and through said channels thereby to perfuse the samples with media, andmeasuring one or more times the concentration of one or more solutes in media about said samples in the plurality of said wells. 25. The method of claim 17 comprising the additional step of placing one or a plurality of beads bearing immobilized binders for biomolecules of interest secreted from or absorbed by the sample in the media and using the beads to detect the presence or concentration of said biomolecules in the microenvironment of the sample.

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IPC	Description
A	생활필수품
A62	인명구조; 소방(사다리 E06C)
A62B	인명구조용의 기구, 장치 또는 방법(특히 의료용에 사용되는 밸브 A61M 39/00; 특히 물에서 쓰이는 인명구조 장치 또는 방법 B63C 9/00; 잠수장비 B63C 11/00; 특히 항공기에 쓰는 것, 예. 낙하산, 투출좌석 B64D; 특히 광산에서 쓰이는 구조장치 E21F 11/00)
A62B-1/08	.. 윈치 또는 풀리에 제동기구가 있는 것

내보내기 구분	파일저장 인쇄 메일전송
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Apparatus and methods for three-dimensional tissue measurements based on controlled media flow 원문보기

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Apparatus and methods for three-dimensional tissue measurements based on controlled media flow 원문보기

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