Automated system for cultivating transgenic C. elegans
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12M-001/36
C12M-003/00
C12M-001/12
C12M-001/32
C12M-003/06
C12M-001/34
C12N-005/07
C12M-001/26
C12M-001/00
출원번호
US-0292084
(2016-10-12)
등록번호
US-10184104
(2019-01-22)
발명자
/ 주소
Lianides, Alexander Lee
Lianides, Christopher Lee
출원인 / 주소
Lianides, Alexander Lee
인용정보
피인용 횟수 :
0인용 특허 :
1
초록▼
The present invention provides a self-draining well plate cassette and an automated system for cultivating C. elegans comprising a housing, a well plate assembly having at least two of the self-draining well plate cassettes, a liquid dispensing assembly operated by a first three axes positioner, a w
The present invention provides a self-draining well plate cassette and an automated system for cultivating C. elegans comprising a housing, a well plate assembly having at least two of the self-draining well plate cassettes, a liquid dispensing assembly operated by a first three axes positioner, a wash and imaging assembly operated by a second three axis positioner, a reagent assembly, a pipette tip holder, and a controller. The present invention further provides a method of using the above-described system to cultivate C. elegans.
대표청구항▼
1. A method of using an automated system for cultivating C. elegans comprising: (a) providing an automated system comprising a housing, a well plate assembly having a self-draining well plate cassette, a liquid dispensing assembly operated by a first three axes positioner, a wash and imaging assembl
1. A method of using an automated system for cultivating C. elegans comprising: (a) providing an automated system comprising a housing, a well plate assembly having a self-draining well plate cassette, a liquid dispensing assembly operated by a first three axes positioner, a wash and imaging assembly operated by a second three axis positioner, a reagent assembly, a pipette tip holder, and a controller wherein: (i) the cassette includes a lid, a top plate having open wells, a bottom plate having raised well bottoms and sealing features corresponding to locations of the open wells, a spring loaded connector having biasing tension spring and a lever that controls compression and expansion of the biasing tension spring, a self-sealing drainage valve that only drains out fluid upon coupling with a drainage coupler, and culture inserts;(ii) each of the open wells include a top opening and a bottom opening;(iii) upon compression of the biasing tension spring by the lever, the bottom openings of the open wells couple with the sealing features of the well bottoms to form sealed wells;(iv) upon expansion of the biasing tension spring by the lever, the bottom openings decouple from the sealing features of well bottoms allowing any fluid contained in each of the sealed wells of the cassette to leak out onto the bottom plate and to drain out via the drainage valve when the drainage valve is coupled to a drainage coupler of the wash and imaging assembly;(v) the well plate assembly further includes a first orbital shaker and a support structure having multiple trays and a support base;(vi) the wash and imaging assembly includes an automated buffer dispensing head designed to accept pipette tips, a first cassette grabber, a camera, a washing platform, a linear actuator having a switch controller, and the drainage coupler;(vii) the liquid dispensing assembly includes an automated aspiring and dispensing pipette, a liquid dispensing lid sensor, a liquid level sensor, and a second cassette grabber;(viii) the reagent assembly includes a container rack, at least one reagent container containing a bacterial solution, a reagent platform, and a second orbital shaker activated to shake the at least one reagent container; and(ix) the controller controls operations of the first three axes positioner, the second three axis positioner, the first orbital shaker, the second orbital shaker, the liquid dispensing assembly, the wash and imaging assembly, and the reagent assembly;(b) placing the cassette onto one of the multiple trays of the support structure of the well plate assembly wherein the culture inserts containing C. elegans are bathing in bacterial solutions within the seal wells of the cassette;(c) activating the first orbital shaker to shake the bacterial solutions;(d) deactivating the first orbital shaker;(e) activating the first cassette grabber to transfer the cassette from the support structure to a first predetermined position on the washing platform wherein the drainage valve couples with the drainage coupler, and the lever engages with the switch controller allowing the linear actuator to control the lever's ability to cause compression or expansion of the biasing tension spring;(f) activating the first cassette grabber to transfer the lid from the cassette to a second predetermined position on the washing platform;(g) activating the linear actuator to cause expansion of the biasing tension spring by the lever resulting in decoupling of the bottom openings from the sealing features and leakage of the bacterial solutions out of the cassette via the drainage valve and the drainage coupler;(h) activating the automated buffer dispensing head to dispense buffer solution into the culture inserts allowing the buffer solution to flow through the culture inserts thereby cleaning the C. elegans and removing waste from the culture inserts;(i) activating the linear actuator to cause compression of the biasing tension spring by the lever resulting in coupling of the bottom openings with the sealing features to form the sealed wells containing the culture inserts with the C. elegans and the buffer solution;(j) deactivating the automated buffer dispensing head to stop the dispensation of the buffer solution into the culture inserts;(k) imaging the C. elegans with the camera;(l) activating the linear actuator to cause expansion of the biasing tension spring by the lever resulting in decoupling of the bottom openings from the sealing features and leakage of the buffer solution onto the bottom plate and drainage of the buffer solution out of the cassette via the drainage valve and the drainage coupler(m) activating the linear actuator to cause compression of the biasing tension spring by the lever resulting in coupling of the bottom openings with the sealing features to form the sealed wells containing the culture inserts with the C. elegans; (n) activating the first cassette grabber to transfer the lid from the second predetermined position on the washing platform back to the cassette;(o) activating the first cassette grabber to transfer the cassette from the washing platform to one of the multiple trays of the support structure;(p) activating the second cassette grabber to transfer the cassette from the support structure to a third predetermined position on the reagent platform;(q) activating the second cassette grabber to transfer the lid from the cassette to a fourth predetermined position on the reagent platform;(r) deactivating the second orbital shaker;(s) activating the automated aspiring and dispensing pipette of the liquid dispensing assembly to transfer a predetermined amount of bacterial solution from the reagent assembly to each of the sealed wells of the cassette;(t) activating the second orbital shaker;(u) activating the second cassette grabber to transfer the lid from the fourth predetermined position on the reagent platform back to the cassette;(v) activating the second cassette grabber to transfer the cassette from the reagent platform to one of the multiple trays of the support structure; and(w) activating the first orbital shaker. 2. The method of claim 1 wherein the automated system further includes one or more additional self-draining well plate cassettes; and the method further includes repeating steps (b) through (w) for each of the additional cassettes. 3. The method of claim 1 wherein the steps (b) through (w) are repeated multiple times until the C. elegans have reached a desired cultivated growth stage. 4. The method of claim 1 wherein diameter of the bottom opening is smaller than diameter of the top opening. 5. The method of claim 1 wherein the sealing feature is a silicone surface. 6. The method of claim 1 wherein the sealing feature is at least one O-ring. 7. The method of claim 1 wherein the cassette further includes a culture insert carrying plate. 8. The method of claim 1 wherein the method is used to cultivate transgenic C. elegans. 9. The method of claim 1 wherein: (i) the at least one reagent container includes an automated lid controlled by a reagent lid sensor and an automated hinge;(ii) when the reagent lid sensor receives a first predetermined signal from the liquid dispensing lid sensor, the reagent lid sensor activates the automated hinge to open the automated lid; and(iii) when the reagent lid sensor receives a second predetermine signal from the liquid dispensing lid sensor, the reagent lid sensor deactivates the automated hinge in order to close the automated lid. 10. A method of using an automated system for cultivating C. elegans comprising: (a) providing an automated system comprising a housing, a well plate assembly having more than one self-draining cassettes, a liquid dispensing assembly operated by a first three axes positioner, a wash and imaging assembly operated by a second three axis positioner, a reagent assembly, a pipette tip holder, and a controller wherein: (i) each of the cassettes includes a lid, a top plate having open wells, a bottom plate having raised well bottoms and sealing features corresponding to locations of the open wells, a spring loaded connector having biasing tension spring and a lever that controls compression and expansion of the biasing tension spring, a self-sealing drainage valve that only drains out fluid upon coupling with a drainage coupler, and culture inserts;(ii) each of the open wells includes a top opening and a bottom opening;(iii) upon compression of the biasing tension spring by the lever, the bottom openings of the open wells couple with the sealing features of the well bottoms to form sealed wells;(iv) upon expansion of the biasing tension spring by the lever, the bottom openings decouple from the sealing features of well bottoms allowing any fluid contained in each of the sealed wells to leak out onto the bottom plate and to drain out via the drainage valve when the drainage valve is coupled to a drainage coupler of the wash and imaging assembly;(v) the well plate assembly further includes a first orbital shaker and a support structure having multiple trays and a support base;(vi) the wash and imaging assembly includes an automated buffer dispensing head designed to accept pipette tips, a first cassette grabber, a camera, a washing platform, a linear actuator having a switch controller, and the drainage coupler;(vii) the liquid dispensing assembly includes an automated aspiring and dispensing pipette, a liquid dispensing lid sensor, a liquid level sensor, and a second cassette grabber;(viii) the reagent assembly includes a container rack, at least one reagent container containing a bacterial solution, a reagent platform, and a second orbital shaker activated to shake the at least one reagent container; and(ix) the controller controls operations of the first three axes positioner, the second three axis positioner, the first orbital shaker, the second orbital shaker, the liquid dispensing assembly, the wash and imaging assembly, and the reagent assembly;(b) placing the cassettes onto the multiple trays of the support structure of the well plate assembly wherein the culture inserts containing C. elegans are bathing in bacterial solutions within the seal wells of each of the cassettes and performing the following steps for each of the cassettes;(c) activating the first orbital shaker to shake the bacterial solutions;(d) deactivating the first orbital shaker;(e) activating the first cassette grabber to transfer one of the cassettes from the support structure to a first predetermined position on the washing platform wherein the drainage valve couples with the drainage coupler, and the lever engages with the switch controller allowing the linear actuator to control the lever's ability to cause compression or expansion of the biasing tension spring;(f) activating the first cassette grabber to transfer the lid from the cassette to a second predetermined position on the washing platform;(g) activating the linear actuator to cause expansion of the biasing tension spring by the lever resulting in decoupling of the bottom openings from the sealing features and leakage of the bacterial solutions out of the cassette via the drainage valve and the drainage coupler;(h) activating the automated buffer dispensing head to dispense buffer solution into the culture inserts allowing the buffer solution to flow through the culture inserts thereby cleaning the C. elegans and removing waste from the culture inserts;(i) activating the linear actuator to cause compression of the biasing tension spring by the lever resulting in coupling of the bottom openings with the sealing features to form the sealed wells containing the culture inserts with the C. elegans and the buffer solution;(j) deactivating the automated buffer dispensing head to stop the dispensation of the buffer solution into the culture inserts;(k) imaging the C. elegans with the camera;(l) activating the linear actuator to cause expansion of the biasing tension spring by the lever resulting in decoupling of the bottom openings from the sealing features and leakage of the buffer solution onto the bottom plate and drainage of the buffer solution out of the cassette via the drainage valve and the drainage coupler(m) activating the linear actuator to cause compression of the biasing tension spring by the lever resulting in coupling of the bottom openings with the sealing features to form the sealed wells containing the culture inserts with the C. elegans; (n) activating the first cassette grabber to transfer the lid from the second predetermined position on the washing platform back to the cassette;(o) activating the first cassette grabber to transfer the cassette from the washing platform to one of the multiple trays of the support structure;(p) activating the second cassette grabber to transfer the cassette from the support structure to a third predetermined position on the reagent platform;(q) activating the second cassette grabber to transfer the lid from the cassette to a fourth predetermined position on the reagent platform;(r) deactivating the second orbital shaker;(s) activating the automated aspiring and dispensing pipette of the liquid dispensing assembly to transfer a predetermined amount of bacterial solution from the reagent assembly to each of the sealed wells of the cassette;(t) activating the second orbital shaker;(u) activating the second cassette grabber to transfer the lid from the fourth predetermined position on the reagent platform back to the cassette;(v) activating the second cassette grabber to transfer the cassette from the reagent platform to one of the multiple trays of the support structure; and(w) activating the first orbital shaker. 11. The method of claim 10 wherein the steps (b) through (w) are repeated multiple times until the C. elegans have reached a desired cultivated growth stage. 12. The method of claim 10 wherein diameter of the bottom opening is smaller than diameter of the top opening. 13. The method of claim 10 wherein the sealing feature is a silicone surface. 14. The method of claim 10 wherein the sealing feature is at least one O-ring. 15. The method of claim 10 wherein each of the cassettes further includes a culture insert carrying plate. 16. The method of claim 10 wherein: (i) the at least one reagent container includes an automated lid controlled by a reagent lid sensor and an automated hinge;(ii) when the reagent lid sensor receives a first predetermined signal from the liquid dispensing lid sensor, the reagent lid sensor activates the automated hinge to open the automated lid; and(iii) when the reagent lid sensor receives a second predetermine signal from the liquid dispensing lid sensor, the reagent lid sensor deactivates the automated hinge in order to close the automated lid. 17. The method of claim 10 wherein the method is used to cultivate transgenic C. elegans.
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이 특허에 인용된 특허 (1)
Malin, Cosmas G., Device for storing and handling Petri dishes, storage device and storage slot for laboratory objects.
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