Materials and methods for cell-based assays
First Claim
1. A device for performing dynamic cellular assays utilizing a microplate reader, the device comprising:
- a plate having a plurality of vessels, each vessel having a bottom and at least one wall extending around said bottom to define a cavity operable to hold a fluid therein, the cavity having a first area and a second area, the first area and the second area lying on a common horizontal plane normal to a detected emission direction, and the second area is at least about three times larger than the first area;
a plurality of cell support structures, each cell support structure having a porosity operable to permit transmission of at least one form of radiant energy therethrough, and having a thickness operable to allow three-dimensional cell growth therein and to allow amplification of the radiant energy through tunneling therethrough,wherein a cell support structure is anchored in the first area of the cavity of each vessel such that when the cavity is filled or substantially filled with a fluid, the fluid contacts the cell support structure in the first area of the cavity and the second area of the cavity is substantially occupied by the fluid;
wherein an emission from cells in the cell support structure is intensified in and can be read from the first area of the cavity;
wherein a noise signal resulting substantially from the fluid can be read from the second area of the cavity, the noise signal useful for performing noise correction of the emission read from the first area of the cavity.
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Accused Products
Abstract
The methods generally relate to three-dimensional culturing of cells with the steps of seeding a cell support structure in a device with at least one vessel for cell culture having a bottom, at least one wall and a cell support structure for three-dimensional cell culture. The cell support structure is inside the at least one vessel and the device is configured such that assays can be performed. Devices for performing cell culture and assays are also provided. It will be understood that the devices can have many different suitable configurations. In general, the devices are selected such that they can be used with suitable radiation based assays.
28 Citations
25 Claims
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1. A device for performing dynamic cellular assays utilizing a microplate reader, the device comprising:
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a plate having a plurality of vessels, each vessel having a bottom and at least one wall extending around said bottom to define a cavity operable to hold a fluid therein, the cavity having a first area and a second area, the first area and the second area lying on a common horizontal plane normal to a detected emission direction, and the second area is at least about three times larger than the first area; a plurality of cell support structures, each cell support structure having a porosity operable to permit transmission of at least one form of radiant energy therethrough, and having a thickness operable to allow three-dimensional cell growth therein and to allow amplification of the radiant energy through tunneling therethrough, wherein a cell support structure is anchored in the first area of the cavity of each vessel such that when the cavity is filled or substantially filled with a fluid, the fluid contacts the cell support structure in the first area of the cavity and the second area of the cavity is substantially occupied by the fluid; wherein an emission from cells in the cell support structure is intensified in and can be read from the first area of the cavity; wherein a noise signal resulting substantially from the fluid can be read from the second area of the cavity, the noise signal useful for performing noise correction of the emission read from the first area of the cavity. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method for analyzing cellular function of cells in three-dimensional culture, the method comprising:
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seeding at least one cell support structure with at least one cell, the cell support structure having a porosity operable to permit transmission of at least one form of radiant energy therethrough, and having a thickness operable to allow three-dimensional cell growth therein, providing a plate having a plurality of vessels, each vessel having a bottom and at least one wall extending around said bottom to define a cavity operable to hold a fluid therein, the cavity having a first area and a second area, the first area and the second area lying on a common horizontal plane normal to a detected emission direction, and the second area is at least about three times larger than the first area, anchoring a cell support structure in the first area of the cavity of each vessel; disposing a fluid in each vessel to culture the at least one cell such that the first area of the cavity is substantially occupied by the cell support structure and the fluid, and the second area of the cavity is substantially occupied by the fluid; detecting an emission from the culture in the first area of the cavity with a microplate reader; detecting a noise signal from the fluid in the second area of the cavity with the microplate reader; and using the noise signal to perform noise correction of the emission. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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Specification