Assay apparatus and methods
First Claim
1. An apparatus for exposing cells to a test compound solution to determine whether the test compound solution induces progressive cell activity response in the cells, the apparatus comprising:
- a substrate having an inner surface and an outer surface opposite to the inner surface;
a plate disposed over the substrate such that a cavity is defined between the plate and the substrate,wherein the plate has a first surface and a second surface opposite to the first surface, the second surface facing the inner surface of the substrate,wherein the plate defines a through-well extending from the first surface of the plate to the second surface of the plate,wherein the through-well exposes the inner surface of the substrate, andwherein the through-well is in fluid communication with the cavity; and
a hydrophobic barrier on the substrate and/or the plate,wherein the hydrophobic barrier defines a test site enclosing the through-well when the apparatus is viewed from a plan view perpendicular to a plane defined by the plate,wherein the hydrophobic barrier allows passage of gas outside of the test site while limiting passage of liquid outside of the test site,wherein the substrate, the plate, and the hydrophobic barrier are configured to contain the cells within the test site,wherein the through-well is disposed at a first side portion of the test site when viewed from the plan view, the test site having a second side portion opposite to the first side portion,wherein, when viewed from the plan view, the test site has a length from the first side portion to the second side portion, and the length is greater than a width of the test site,wherein the plate and the substrate define a gap between the substrate and an edge of the through-well at the second surface of the plate, on a side of the through-well facing the second side portion of the test site, andwherein the gap extends substantially across the width of the test site.
1 Assignment
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Accused Products
Abstract
Apparatus and methods for determining whether a test compound induces cell activity, changes cell activity, prevents cell activity, or inhibits cell activity. An embodiment comprises placing a test compound solution in contact with a cell suspension media containing cells, diffusing the test compound solution into the cell suspension from one or more sides, and detecting activity in the cells with respect to their distance from the side from which the test compound is diffusing. Embodiments may provide an apparatus that allows a side source, a point source, or both, from which a test compound solution diffuses into a cell suspension media and contacts cells. Detecting cell activity may involve detecting activity in a first cell group proximate to the side from which the test compound is diffusing, and detecting activity in a second cell group farther than the first cell group from the side from which the test compound is diffusing.
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Citations
21 Claims
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1. An apparatus for exposing cells to a test compound solution to determine whether the test compound solution induces progressive cell activity response in the cells, the apparatus comprising:
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a substrate having an inner surface and an outer surface opposite to the inner surface; a plate disposed over the substrate such that a cavity is defined between the plate and the substrate, wherein the plate has a first surface and a second surface opposite to the first surface, the second surface facing the inner surface of the substrate, wherein the plate defines a through-well extending from the first surface of the plate to the second surface of the plate, wherein the through-well exposes the inner surface of the substrate, and wherein the through-well is in fluid communication with the cavity; and a hydrophobic barrier on the substrate and/or the plate, wherein the hydrophobic barrier defines a test site enclosing the through-well when the apparatus is viewed from a plan view perpendicular to a plane defined by the plate, wherein the hydrophobic barrier allows passage of gas outside of the test site while limiting passage of liquid outside of the test site, wherein the substrate, the plate, and the hydrophobic barrier are configured to contain the cells within the test site, wherein the through-well is disposed at a first side portion of the test site when viewed from the plan view, the test site having a second side portion opposite to the first side portion, wherein, when viewed from the plan view, the test site has a length from the first side portion to the second side portion, and the length is greater than a width of the test site, wherein the plate and the substrate define a gap between the substrate and an edge of the through-well at the second surface of the plate, on a side of the through-well facing the second side portion of the test site, and wherein the gap extends substantially across the width of the test site. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for determining whether a test compound solution induces progressive cell activity response, the method comprising:
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depositing a media containing cells into a cavity defined between a plate and a substrate, wherein the substrate has an inner surface and an outer surface opposite to the inner surface, wherein the plate has a first surface and a second surface opposite to the first surface, the second surface facing the inner surface of the substrate, wherein the plate defines a through-well extending from the first surface of the plate to the second surface of the plate, wherein the through-well exposes the inner surface of the substrate, wherein the through-well is in fluid communication with the cavity, wherein a hydrophobic barrier on the substrate and/or the plate defines a test site enclosing the through-well when the substrate and the plate are viewed from a plan view perpendicular to a plane defined by the plate, wherein, when viewed from the plan view, the test site is elongated, having a length greater than a width, wherein the hydrophobic barrier allows passage of gas outside of the test site while limiting passage of liquid outside of the test site, wherein the substrate, the plate, and the hydrophobic barrier are configured to contain the cells within the test site, wherein the through-well is disposed at a first side portion of the test site when viewed from the plan view, the test site having a second side portion opposite to the first side portion, wherein the plate and the substrate define a gap between the substrate and an edge of the through-well at the second surface of the plate, on a side of the through-well facing the second side portion of the test site, and wherein the gap extends substantially across the width of the test site; allowing the media containing cells to wick into the cavity out to the hydrophobic barrier and form a meniscus at the gap; placing the test compound solution in the through-well and in contact with the media containing cells at the meniscus at the gap; diffusing into the media containing cells the test compound solution in a substantially linear front originating from the gap, spanning the width of the test site, and moving across the length of the test site from the first side portion of the test site to the second side portion of the test site, wherein a first cell of the media is closer to the gap than a second cell of the media; allowing the test compound solution to diffuse into the media for a predetermined duration such that the first cell is exposed to the test compound solution longer than the second cell is exposed to the test compound solution; after the predetermined duration, detecting a degree of activity of the first cell and a degree of activity of the second cell; and comparing the degree of activity of the first cell to the degree of activity of the second cell to determine the presence or absence of a progressive cell activity response by the cells of the media to determine whether the test compound solution induces progressive cell activity response. - View Dependent Claims (16, 17, 18, 19, 20, 21)
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Specification