초록 ▼

This invention provides methods and apparatus for performing microanalytic analyses and procedures, particularly miniaturized cell based assays. These methods are useful for performing a variety of cell-based assays, including drug candidate screening, life sciences research, and clinical and molecu

This invention provides methods and apparatus for performing microanalytic analyses and procedures, particularly miniaturized cell based assays. These methods are useful for performing a variety of cell-based assays, including drug candidate screening, life sciences research, and clinical and molecular diagnostics.

대표청구항 ▼

1. A centripetally-motivated microsystems platform comprising:a) a rotatable platform comprising a substrate having a surface comprising a one or a multiplicity of microfluidics structures embedded in the surface of the platform, wherein each microfluidics structure comprisesi) a distribution manifo

1. A centripetally-motivated microsystems platform comprising:a) a rotatable platform comprising a substrate having a surface comprising a one or a multiplicity of microfluidics structures embedded in the surface of the platform, wherein each microfluidics structure comprisesi) a distribution manifold,i) one or a plurality of cell culture chambers adapted or treated to permit cell attachment and growth, andii) one or a plurality of overflow reservoirswherein each of said cell culture chambers is fluidly connected to the distribution manifold and to at least one of the plurality of overflow reservoirs microchannels, and wherein the platform further comprisesb) a distribution manifold overflow reservoir, wherein the overflow reservoir is fluidly connected to the distribution manifold by a microchannel that makes a fluid connection with the distribution manifold at a position on the manifold proximal to the axis of rotation,and wherein fluid within the microchannels of the platform is moved through said microchannels by centripetal force arising from rotational motion of the platform for a time and a rotational velocity sufficient to move the fluid through the microchannels. 2. A microsystems platform according to claim 1, wherein each of the plurality of cell culture chambers further comprises a cell support or filtering element. 3. A microsystems platform according to claim 1, further comprising a plurality of detection chambers, wherein each detection chamber is fluidly connected by microchannels to one of the plurality of cell culture chambers and one of the plurality of overflow reservoirs, and is positioned on the platform between the cell culture chamber and the overflow reservoir. 4. A microsystem platform of claim 3 wherein the detection reservoirs are optically transparent. 5. A microsystem platform of claim 1 wherein the distribution manifold has a volumetric capacity of from about 1 nL to about 500 μL. 6. A microsystem platform of claim 1 wherein each cell culture chamber has a volumetric capacity of from about 2 nL to about 1000 μL. 7. A microsystem platform of claim 3 wherein each detection reservoir has a volumetric capacity of from about 2 nL to about 1000 μL. 8. A microsystem platform of claim 2 wherein the cell support or filtering element is a porous membrane having a pore size that prevents passage of cultured cells therethrough. 9. A microsystem platform of claim 1 comprising from about 24 to about 10,000 microfluidics structures. 10. A microsystem platform of claim 1 that is a circular disk having a radius of about 1 to about 25 cm. 11. The microsystem platform of claim 1, wherein the microsystem platform is constructed of a material selected from the group consisting of an organic material, an inorganic material, a crystalline material and an amorphous material. 12. The microsystem platform of claim 11, wherein the microsystem platform further comprises a material selected from the group consisting of silicon, silica, quartz, a ceramic, a metal or a plastic. 13. The microsystem platform of claim 1, wherein the microsystem platform has a thickness of about 0.1 to 100 mm, and wherein the cross-sectional dimension of the microchannels embedded therein is less than 1 mm and from 1 to 90 percent of said cross-sectional dimension of the platform. 14. A microsystem platform of claim 3 wherein each detection chamber has a volumetric capacity of from about 2 nL to about 1000 μL. 15. A microsystems platform of claim 1, wherein each of the microchannels fluidly connected to the cell culture chamber and overflow reservoir is coated with a hydrophobic coating. 16. The microsystem platform of claim 1, comprising a first layer and a second layer, wherein the first layer comprises a distribution manifold and cell culture chambers, and the second layer comprises microchannels, distribution manifold overflow reservoir, detection chambers and overflow reservoirs, wherein the distribution manifold and cell culture chambers in the first layer are fluidly connected by the microchannels, distribution manifold overflow reservoir, detection chambers and overflow reservoirs in the second layer when the first layer is in contact with the second layer. 17. A centripetally-motivated fluid micromanipulation apparatus that is a combination ofa microsystem platform according to claim 1, anda micromanipulation device, comprising a base, a rotating means, a power supply and user interface and operations controlling means, wherein the rotating means is operatively linked to the microsystem platform and in rotational contact therewithwherein a volume of a fluid within the microchannels of the platform is moved through said microchannels by centripetal force arising from rotational motion of the platform for a time and a rotational velocity sufficient to move the fluid through the microchannels. 18. The apparatus of claim 17, wherein the rotating means of the device is a motor. 19. The apparatus of claim 17, wherein the device comprises a rotational motion controlling means for controlling the rotational acceleration and velocity of the microsystem platform. 20. An apparatus of claim 17 wherein the micromanipulation apparatus further comprises an optical detector that measures absorbance, fluorescence, epifluorescence or chemoluminescence. 21. An apparatus of claim 17 wherein the micromanipulation apparatus further comprises a scanning, imaging, or confocal microscopy detector. 22. An apparatus of claim 17 wherein the micromanipulation apparatus further comprises a radiometric detector. 23. An apparatus of claim 20, wherein the detector is brought into alignment with the collection chamber on the platform by rotational motion of the microsystem platform. 24. The apparatus of claim 23, wherein the detector is an optical detector comprising a light source and a photodetector. 25. A method for performing a cell-based assay, comprising the steps of:a) applying a volume of a fluid comprising a cell suspension to the distribution manifold of a microsystem platform of claim 1 when the platform is stationary,b) rotating the platform at a first rotational speed wherein a portion of the suspension occupies one or a plurality of subvolumes of the distribution manifold,c) rotating the platform at a second rotational speed that is higher than the first rotational speed wherein the portion of the cell suspension that does not occupy one or a plurality of subvolumes of the distribution manifold is motivated from the distribution manifold to the cell suspension overflow reservoir;d) rotating the platform at a third rotational speed that is higher than the second rotational speed to motivate fluid flow from the subvolumes of the distribution manifold into one or each of a plurality of cell culture chambers;e) incubating the platform for a time and under conditions for the cells to attach to the cell culture chamber and grow;f) applying a volume of a reagent solution to the distribution manifold of a microsystem platform of claim 1 when the platform is stationary,g) rotating the platform at a first rotational speed wherein a portion of the reagent solution occupies one or a plurality of subvolumes of the distribution manifold,h) rotating the platform at a second rotational speed that is higher than the first rotational speed wherein the portion of the reagent solution that does not occupy one or a plurality of subvolumes of the distribution manifold is motivated from the distribution manifold to the cell suspension overflow reservoir;i) rotating the platform at a third rotational speed that is higher than the second rotational speed to motivate fluid flow from the subvolumes of the distribution manifold into one or each of a plurality of cell culture chambers;j) incubating the platform for a time and under conditions for the cells to interact with the reagent and produce a detectable product; andk) detecting a product of the biological or biochemical reactio n. 26. A method according to claim 22, wherein the reagent is a drug lead compound.