Apparatus and method of preparation for automated high output biopolymer crystallization via vapor diffusion sitting drop and micro-batch techniques
First Claim
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1. A robotic station for protein crystallization comprising:
- (a) a magazine storing at least one empty microplate, (b) said microplate being comprised of an array of wells paired with reservoirs;
(c) a nest that holds the microplate in position and indexes via an electrically or mechanically driven mechanism;
(d) a mechanism for feeding a microplate into said nest;
(e) a liquid dispensing system with at least one probe, each said probe containing at least one bore that delivers specified volumes into specific wells and reservoirs of said microplate;
(f) an electrically or mechanically driven system that positions said probe over targeted reservoirs and wells;
(g) a sealing mechanism that transfers a seal from a dispenser onto selected wells and reservoirs to seal them from the environment;
(h) an incubator for storing the microplates in a controlled environment;
(i) a microplate removing mechanism that removes a microplate from the nest and places it in said incubator;
(j) a stacking mechanism that stacks the microplates inside said incubator.
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Abstract
A fully automated, high output robotic station for the crystallization of proteins and other biopolymers via the sitting drop vapor diffusion and micro-batch methods. This station performs microplate feeding, liquid aspiration, dilution, and dispensing, as well as microplate sealing and storage into a temperature controlled hotel. All functions are microprocessor controlled. The operator designs the process via graphical user interface, after which the station executes the process automatically to completion.
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Citations
16 Claims
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1. A robotic station for protein crystallization comprising:
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(a) a magazine storing at least one empty microplate, (b) said microplate being comprised of an array of wells paired with reservoirs;
(c) a nest that holds the microplate in position and indexes via an electrically or mechanically driven mechanism;
(d) a mechanism for feeding a microplate into said nest;
(e) a liquid dispensing system with at least one probe, each said probe containing at least one bore that delivers specified volumes into specific wells and reservoirs of said microplate;
(f) an electrically or mechanically driven system that positions said probe over targeted reservoirs and wells;
(g) a sealing mechanism that transfers a seal from a dispenser onto selected wells and reservoirs to seal them from the environment;
(h) an incubator for storing the microplates in a controlled environment;
(i) a microplate removing mechanism that removes a microplate from the nest and places it in said incubator;
(j) a stacking mechanism that stacks the microplates inside said incubator. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method of preparing microplates for vapor diffusion crystallization via sitting drop method , comprising of the following steps:
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(a) feeding a microplate from a magazine into a nest;
(b) reading the code of the microplate;
(c) indexing the nest into proper position to allow selected wells and reservoirs of the microplate to be injected with ingredients via the transport of fluid through transport lines into a probe for dispensing said fluid;
(d) flushing the fluid transport lines and drying them;
(e) washing and drying the probe(s);
(f) positioning the probe(s) over a subset of reservoirs for dispensing specified reagent;
(g) repetitive dispensing and flushing of reagents and washing and drying of lines and probe(s) until said subset of reservoirs contain all necessary ingredients;
(h) loading probe(s) with a biopolymer to be crystallized;
(i) positioning of probe(s) over the wells corresponding to the said subset of reservoirs and dispensing solution that contains the biopolymer to be crystallized;
(j) alternating washing and drying with positioning of probe(s) over each reservoir of said subset and dispensing a small portion of the contents of said reservoir into a corresponding well;
(k) sealing said subset of wells and reservoirs;
(l) repetitive dispensing of ingredients into subsequent subsets of reservoirs and wells and flushing and drying of lines and probe(s) and sealing said reservoirs and wells until said microplate is completely full and covered;
(m) removing said completed microplate from the nest and loading said microplate into an incubator;
(n) indexing said incubator to accommodate next microplate;
(o) repeating all previous steps until all microplates are completed.
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16. A method of preparing microplates using a multibore probe and the micro-batch crystallization technique, comprising the following steps:
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(a) feeding a microplate from a magazine to a nest;
(b) reading the microplate code;
(c) indexing the nest into proper position to allow the reservoirs of the microplate to be filled with ingredients via the transport of fluid through transport lines into said probe for dispensing said fluid into said microplate;
(d) flushing the fluid transport lines with water and drying them with nitrogen gas or compressed air;
(e) washing and drying the probe;
(f) positioning the probe over a subset of reservoirs that are computer selected and dispensing paraffin oil into each of them;
(g) loading each bore of the multibore probe with a solution comprising one ingredient or biopolymer;
(h) positioning the probe into an oil filled reservoir and dispensing a specified volume of solution;
(i) retracting the probe from the oil;
(j) repeating said positioning, dispensing, and retraction until all reservoirs of said subset that contain the same combination of ingredients are completed;
(k) repetitive dispensing and flushing of reagents and flushing and drying of lines and probe until said subset of reservoirs contain all necessary ingredients;
(l) feeding tape from a dispenser, cutting it, and placing it over said subset of wells and reservoirs;
(m) repetitive dispensing of ingredients into subsequent subsets of reservoirs and wells and flushing and drying of the lines and the probe and sealing of reservoirs and wells with tape until said microplate is completely full and covered;
(n) removing the completed microplate from said nest and loading said microplate into an incubator;
(o) indexing said hotel downward to accommodate the next microplate;
(p) repeating all previous steps until all microplates are completed.
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Specification