Automated semi-solid matrix assay and liquid handler apparatus for the same
First Claim
1. An improved liquid handling machine for automatically transferring liquid between a plurality of liquid containing wells, said machine having:
- a head;
a plunger assembly mounted on said head, said plunger assembly including a plurality of pipettes having depending ends for receiving tips, a plurality of plungers respectively disposed within said pipettes, said plungers being movable coaxially within said pipettes to vary their internal volumes;
a tip ejector for removing tips disposed on said depending ends of said pipettes;
a support mounted beneath said head, said support having at least a first work station being adapted to accommodate a tray having pipette tip-receiving receptacles, a second and a third work station each being adapted to accommodate a respective tray of said liquid containing wells;
said support and said head being movable relatively to one another to place said pipettes in registry with any of said tips in said tip-receiving receptacles and any of said wells in registry with said pipettes;
a motion controller for controlling the relative motion between said head and said support as well as the motion of said plungers and said tip ejector to effect transfer of liquid between the liquid containing wells and to replace tips on the ends of said pipettes with other tips disposed in at least some of the pipette tip-receiving receptacles between predetermined liquid transfer steps, wherein the improvement comprises;
said support having a plurality of temperature control elements for independently controlling the temperature at each of said second and third work stations to maintain said liquid in said liquid containing wells at a predetermined temperature, said support also having a sensor at each of said second and third work stations for sensing the temperature, and a temperature controller for controlling said temperature control elements and said sensors.
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Accused Products
Abstract
An improved liquid handling machine capable of regulating the temperature of assay compounds in the automated preparation of culture trays for biological assays is disclosed. The machine includes a horizontally movable table positioned beneath a vertically movable head. The table is divided into a plurality of stations holding mixing trays, culture trays and reservoirs of liquid assay compound. The head holds a plurality of pipettes which aspirate and expel liquid to transfer and mix the assay compounds between the reservoir, the mixing trays and the culture trays upon coordinated movement of the head and the table as controlled by a microprocessor. Each station on the table has independent heating, cooling and temperature sensing elements for regulating the temperature of the liquid held in a tray or reservoir at the station. A device for automatically evaluating the results of the assay, such as by fluorescence, spectrophotometric or radioactive techniques is incorporated with the improved liquid handling machine.
81 Citations
99 Claims
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1. An improved liquid handling machine for automatically transferring liquid between a plurality of liquid containing wells, said machine having:
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a head;
a plunger assembly mounted on said head, said plunger assembly including a plurality of pipettes having depending ends for receiving tips, a plurality of plungers respectively disposed within said pipettes, said plungers being movable coaxially within said pipettes to vary their internal volumes;
a tip ejector for removing tips disposed on said depending ends of said pipettes;
a support mounted beneath said head, said support having at least a first work station being adapted to accommodate a tray having pipette tip-receiving receptacles, a second and a third work station each being adapted to accommodate a respective tray of said liquid containing wells;
said support and said head being movable relatively to one another to place said pipettes in registry with any of said tips in said tip-receiving receptacles and any of said wells in registry with said pipettes;
a motion controller for controlling the relative motion between said head and said support as well as the motion of said plungers and said tip ejector to effect transfer of liquid between the liquid containing wells and to replace tips on the ends of said pipettes with other tips disposed in at least some of the pipette tip-receiving receptacles between predetermined liquid transfer steps, wherein the improvement comprises;
said support having a plurality of temperature control elements for independently controlling the temperature at each of said second and third work stations to maintain said liquid in said liquid containing wells at a predetermined temperature, said support also having a sensor at each of said second and third work stations for sensing the temperature, and a temperature controller for controlling said temperature control elements and said sensors. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. An assay method determining the effect of a test compound on a cell in a predetermined well comprising semi-solid matrix, comprising:
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(a) using a liquid handler with a first reservoir and a second reservoir to transfer a liquid semi-solid matrix from the first reservoir to a predetermined well of an assay plate;
(b) using a liquid handler to transfer cells from a second reservoir to the predetermined well of an assay plate;
(c) allowing the liquid semi-solid matrix in the predetermined well to solidify;
(d) incubating the cells in the plate for a period of time for the cells to grow into a colony;
(e) adding a predetermined amount of a test compound to the predetermined well; and
(f) determining the biological effect of the test compound on the colony. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80)
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36. An assay method for determining the effect of a test compound on cells comprising:
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(a) using a liquid handler to transfer a liquid semi-solid matrix from a first reservoir to a predetermined well of an assay plate;
(b) using a liquid handler to transfer growth medium from a second reservior to the predetermined well of the assay plate;
(c) using a liquid handler to transfer cells from a third reservoir to the predetermined well of the assay plate;
(d) allowing the liquid semi-solid matrix in the predetermined well of the assay plate to solidify;
(e) incubating the cells in the predetermined well of the assay plate for a period of time for the cells to grow into a colony;
(f) adding a predetermined amount of a test compound to the predetermined well of the assay plate; and
(g) determining the biological effect of the test compound on the colony.
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37. An assay method for determining the effect of a test compound on cells comprising:
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(a) using a liquid handler to transfer a liquid semi-solid matrix from a first reservoir into a predetermined well of the assay plate;
(b) using a liquid handler to transfer growth medium from a second reservior to the predetermined well of the assay plate;
(c) using a liquid handler to transfer cells from a third reservoir to the predetermined well of the assay plate;
(d) using a liquid handler to transfer a predetermined amount of a test compound from a fourth reservoir to the predetermined well of the assay plate;
(e) allowing the liquid semi-solid matrix in the predetermined well of the assay plate to solidify;
(f) incubating the cells in the predetermined well of the assay plate for a period of time for the cells to grow to form a colony; and
(g) determining the biological effect of the test compound on the colony. - View Dependent Claims (38, 39)
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81. A method of using a liquid handler for automated preparation of an assay plate comprising wells with a semi-solid matrix, wherein
the assay plate comprises multiple wells in one or more columns, wherein at least one well is capable of holding a semi-solid matrix, the liquid handler comprises a first reservoir comprising growth medium at one concentration, second reservoir comprising a liquid semi-solid matrix, third reservoir comprising cells in a liquid, and fourth reservoir, and the means to transfer a liquid from a reservoir to a predetermined well in an assay plate, by a) transferring a predetermined amount of growth medium from the first reservoir to the fourth reservoir, b) transferring a predetermined amount of the liquid semi-solid matrix from the second reservoir to the fourth reservoir, c) transferring a predetermined amount of cells in a liquid from the third reservoir to the fourth reservoir, d) mixing the growth medium, liquid semi-solid matrix and cells in the fourth reservoir, e) transferring a predetermined amount of the mixed growth medium, liquid semi-solid matrix and cells from the fourth reservoir into a predetermined well of the first assay plate, and f) allowing the semi-solid matrix to solidify, wherein steps a-c may be performed in any order.
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90. A method of using a liquid handler for automated preparation of an assay plate comprising wells with a semi-solid matrix, wherein
the assay plate comprises multiple wells in one or more columns, wherein at least one well is capable of holding a semi-solid matrix, the liquid handler comprises a first reservoir comprising growth medium, a liquid semi-solid matrix, and cells, and second reservoir, and the means to transfer a liquid from a reservoir to a predetermined well in an assay plate, by a) mixing the growth medium, liquid semi-solid matrix and cells in the first reservoir, b) transferring a predetermined amount of the mixed growth medium, liquid semi-solid matrix and cells from the first reservoir into a predetermined well of the first assay plate, and c) allowing the semi-solid matrix in the predetermined well to solidify, optionally repeating steps a and b, but transferring into a second predetermined well of the first assay plate.
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94. A method of using a liquid handler for automated preparation of an assay plate comprising wells with a semi-solid matrix, wherein
the assay plate comprises multiple wells in one or more columns, wherein at least one well is capable of holding a semi-solid matrix, the liquid handler comprises a first reservoir compriseing growth medium at one concentration, second reservoir comprising a liquid semi-solid matrix, third reservoir comprising cells in a liquid, and fourth reservoir, and the means to transfer a liquid from a reservoir to a predetermined well in an assay plate, by forming a first layer of a semi-solid matrix in a predetermined well of a first assay plate, comprising, by forming a second layer of a semi-solid matrix, growth medium and cells in a predetermined well of a first assay plate, comprising,a) transferring a predetermined amount of the liquid semi-solid matrix from the second reservoir to a predetermined well in the assay plate, b) allowing the semi-solid matrix to solidify, a) transferring a predetermined amount of growth medium from the first reservoir to the fourth reservoir, b) transferring a predetermined amount of the liquid semi-solid matrix from the second reservoir to the fourth reservoir, c) transferring a predetermined amount of cells in a liquid from the third reservoir to the fourth reservoir, d) mixing the growth medium, liquid semi-solid matrix and cells in the fourth reservoir, c) transferring a predetermined amount of the mixed growth medium, liquid semi-solid matrix and cells from the fourth reservoir into a predetermined well of the first assay plate, and f) allowing the second layer of liquid semi-solid matrix to solidify, optionally repeating steps 1 and/or 2, but transferring into a second predetermined well of the first assay plate.
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95. A method for using a liquid handler for automated preparation of an assay plate containing semi-solid matrix, wherein
each plate has at least one well in one or more columns capable of holding a semi-solid matrix, the liquid handler comprises a first reservoir comprising a liquid growth medium in one concentration, a second reservoir a liquid growth medium in second concentration, a third reservoir comprising a liquid growth medium in third concentration, a fourth reservoir containing liquid semi-solid matrix, and a fifth reservoir comprising cells in liquid growth medium at the first concentration; - by
1) preparing a first layer of semi-solid matrix in a predetermined well in a first assay plate, comprising a) transferring a predetermined amount of growth medium from the third reservoir to the liquid semi-solid matrix in a fourth reservoir, to form a liquid mixture, b) transferring a predetermined amount of liquid mixture from the fourth reservoir to a predetermined well in first assay plate, c) allowing the liquid mixture in the predetermined well in the first assay plate to solidify, d) optionally repeating steps b and c with a second predetermined well in the first assay plate;
2) preparing a second layer of semi-solid matrix in a predetermined well in the first assay plate, comprising a) transferring a predetermined amount of the growth medium from the second reservoir to the fifth reservoir comprsing the cells, b) transferring a predetermined amount of liquid semi-solid matrix in the fourth reservoir to the fifth reservoir, c) mixing contents of the fifth reservoir, d) transferring a predetermined amount of the mixure in the fifth reservoir to first layer of semi-solid matrix in a predetermined well in a first assay plate to form a second layer, e) allowing the second layer in the predetermined well to solidify. - View Dependent Claims (96, 97, 98, 99)
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Specification