Virtual wells for use in high throughput screening assays
First Claim
1. A microtiter-like plate comprising:
- a bottom having an upper surface comprising a plurality of virtual wells, said virtual wells being relatively hydrophillic domains within a relatively hydrophobic field; and
a cover or top configured for enclosing said bottom, wherein said cover or top has a lower surface comprising a plurality of virtual wells, and said plurality of virtual wells of said bottom and said top are present in an arrangement such that, when said bottom and said top are in close proximity, column-like fluid wells are formed between said bottom and said top, or fluid is transferred, or components are captured.
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Accused Products
Abstract
Microtiter-like plates containing virtual wells formed by an arrangement of relatively hydrophilic domains within relatively hydrophobic fields are provided. Assay mixtures are confined to the hydrophilic domains of the virtual wells by the edges of the hydrophobic fields. The use of virtual wells allows one to perform homogeneous and capture and wash high throughput screening assays with assay mixtures having volumes on the order of about 100 nl to 10 μl. Virtual wells also provide a means of easily moving fluids, which is particularly useful for simultaneous additions needed for kinetic studies and flash detection and washing. Methods for controlling evaporation during the dispensing of reagents as well as during incubation of high throughput screening utilizing microtiter-like plates containing virtual wells are also provided.
The present invention also provides an inexpensive, disposable device for transferring small volumes of an entire array of compounds from a first microtiter-like plate to a second microtiter-like plate, preserving the spatial arrangement of the compounds. Methods of manufacturing and using the device are also provided.
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Citations
21 Claims
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1. A microtiter-like plate comprising:
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a bottom having an upper surface comprising a plurality of virtual wells, said virtual wells being relatively hydrophillic domains within a relatively hydrophobic field; and
a cover or top configured for enclosing said bottom, wherein said cover or top has a lower surface comprising a plurality of virtual wells, and said plurality of virtual wells of said bottom and said top are present in an arrangement such that, when said bottom and said top are in close proximity, column-like fluid wells are formed between said bottom and said top, or fluid is transferred, or components are captured. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method of transferring fluid to a plurality of virtual wells comprising:
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(a) providing a first plate or lid comprising a plurality of virtual wells to which said fluid is to be transferred;
(b) providing a second plate or lid on which said fluid is present;
(c) moving said first plate or lid and said second plate or lid into close proximity so that fluid is transferred from said second plate or lid to said plurality of virtual wells of said first plate or lid.
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10. A method of adding fluid to a plurality of virtual wells comprising:
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a) providing a plate or lid containing a plurality of virtual wells to which said fluid is to be added;
b) moving said plate or lid into close proximity to a fluid reservoir so that fluid is added to said plurality of virtual wells of said plate or lid from said reservoir; and
c) shaking said plate or lid.
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11. A method of nearly simultaneously adding fluid to a plurality of virtual wells so as to enable detection of flash reagents or to enable kinetic studies comprising:
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(a) providing a plate or lid containing a plurality of virtual wells to which said fluid is to be added;
(b) providing a second plate or lid on which said fluid is present;
(c) moving said first plate or lid and said second plate or lid into close proximity so that all or most of the fluid is transferred from said second plate or lid simultaneously or nearly simultaneously to said plurality of virtual wells of said first plate or mixed with fluid already in said first plate or lid. - View Dependent Claims (12)
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13. A method of removing fluid from a plurality of virtual wells comprising:
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(a) providing a first plate or lid comprising a plurality of virtual wells in which said fluid is present;
(b) providing a second plate or lid onto which said fluid is to be transferred;
(c) moving said first plate or lid and said second plate or lid into close proximity so that fluid is transferred from said plurality of virtual wells of said first plate or lid to said second plate or lid, thus removing some or all of said fluid from said virtual wells of said first plate or lid.
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14. A method for limiting evaporation in a microtiter-like plate containing virtual wells during incubation with a lid comprising providing a fluid reservoir in the plate so that any gas that moves into the plate is humidified before it reaches the virtual wells.
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15. A method of screening using a microtiter-like plate having a top and bottom comprising:
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a) adding a series of reagents to a plurality of virtual wells in a microtiter-like plate;
b) adding a spatially defined array of compounds to the plurality of virtual wells before or after (a);
c) moving said top into close proximity with said bottom so that column-like fluid wells are formed between said bottom and said top, or fluid is transferred, or components are captured;
d) incubating the reagents and compound; and
e) reading a diagnostic signal from the virtual wells. - View Dependent Claims (16, 17, 18)
(i) separating the top and bottom of the microtiter-like plate and adding a new top or bottom that is engineered to bind one or more of the reagents in the virtual wells;
(ii) incubating as desired to allow binding of the reagents to the new top or bottom;
(iii) washing the bound reagents as desired;
(iv) repeating steps (a)-(b) and (i)-(iii) above as desired.
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18. The method of claim 15 comprising repeating step (d) one or more times.
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19. A method of screening to identify a compound capable of modulating a preselected biological activity exhibited by cells comprising:
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(a) providing cells in the virtual wells of a microtiter-like plate, said microtiter-like plate comprising a top and bottom, and a sidewall or spacer that vary in height so that the distance between said bottom and said top is determined by the height of said sidewall or spacer;
(b) exposing the cells to a compound or collection of compounds suspected of being capable of modulating the preselected biological activity to be exhibited by the cells; and
(c) determining whether the preselected biological activity has been modulated. - View Dependent Claims (20)
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21. A method of high throughput screening to identify a substance capable of binding to or modulating the activity of a protein or nucleic acid comprising:
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(a) providing on a first plate or lid said protein or said nucleic acid, in solution, in membranes, or in cells, in virtual wells;
(b) exposing said protein or said nucleic acid to a substance suspected of being capable of binding to or modulating the activity of said protein or said nucleic acid by moving a second plate or lid in close proximity with said first plate or lid, said second plate or lid having said substance disposed thereon; and
(c) determining whether said substance modulates the activity of said protein or said nucleic acid.
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Specification