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US 10,184,104 B2Granted
Automated system for cultivating transgenic C. elegans
OwnerAlexander Lee Lianides, Christopher Lee Lianidesinventors
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Abstract
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.
Claims17
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.
31 more elements
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.
31 more elements
Dependencies are inferred from the claim text.