High throughput cell-based assays fabricated with integrated silicon and cell culture technologies
View Patent ↗The present invention relates to articles and methods involving porous materials (e.g., membranes) which may interact with species, such as biological molecules, cells, etc., whereby the species may adhere to or become immobilized with respect to a surface of the porous material or an adhesion layer coating the porous surface. The porous material may be capable of attaching species with control over the positioning and spatial distribution of the species across the surface of the material. Such articles and methods may be useful in, for example, biological assays, biological sensors, or in the culturing of biological cells.
1. An apparatus comprising:
a membrane, the membrane comprising:
a first side;
a second, opposing side;
a plurality of pores spanning the membrane from the first side to the second side;
a cell adhesive material layer positioned on the first side of the membrane, with the second side of the membrane remaining essentially free of the cell adhesive material; and
cells positioned on the second side of the membrane and attached to the cell adhesive material layer through the pores,
wherein the membrane comprises primarily inorganic material, and
wherein at least about 50% of the pores are positioned less than about 25 microns from a nearest, adjacent pore and the apparatus is configured for cell culturing and/or performing a cell assay.
2. An apparatus as in claim 1 , further comprising:
first and second compartments constructed and arranged to contain fluid, the first and second compartments separated by the membrane; and
components constructed and arranged to facilitate containment, within each compartment, of a fluid positioned adjacent the membrane.
3. An apparatus as in claim 1 , wherein the cell adhesive material layer occludes at least some of the plurality of pores.
4. An apparatus as in claim 3 , wherein the cell adhesive material layer occludes essentially all of the plurality of pores.
5. An apparatus as in claim 1 , wherein the cell adhesive material layer comprises collagen.
6. An apparatus as in claim 1 , wherein the inorganic material is a metal oxide and/or metal nitride.
7. An apparatus as in claim 6 , wherein the inorganic material is silicon nitride.
8. An apparatus as in claim 1 , wherein the membrane comprises at least one portion having a thickness measured between the first and second sides of less than about 30 microns.
9. An apparatus as in claim 8 , wherein the membrane comprises at least one portion having a thickness measured between the first and second sides of less than about 20 microns.
10. An apparatus as in claim 9 , wherein the membrane comprises at least one portion having a thickness measured between the first and second sides of less than about 10 microns.
11. An apparatus as in claim 10 , wherein the membrane comprises at least one portion having a thickness measured between the first and second sides of less than about 5 microns.
12. An apparatus as in claim 1 , wherein no more than about 5% of all pores deviate in size from an average pore size of the plurality of pores by more than about 10%.
13. An apparatus as in claim 1 , wherein no more than about 5% of all pores deviate in size from an average pore size of the plurality of pores by more than about 5%.
14. An apparatus as in claim 1 , wherein the membrane includes at least one portion, having an area of at least about 1 mm 2 , which exhibits at least 30% transmission of light at least one wavelength within a wavelength range of from about 400 nm to about 800 nm.
15. An apparatus as in claim 14 , wherein the membrane includes at least one portion, having an area of at least about 1 mm 2 , which exhibits at least 40% transmission of light at least one wavelength within a wavelength range of from about 400 nm to about 800 nm.
16. An apparatus as in claim 15 , wherein the membrane includes at least one portion, having an area of at least about 1 mm 2 , which exhibits at least 50% transmission of light at least one wavelength within a wavelength range of from about 400 nm to about 800 nm.
17. An apparatus as in claim 1 , wherein each attached cell is associated with a single pore, such that a spatial distribution of the cells on the membrane is directed by a spatial distribution of the pores.
18. An article for cell culturing and/or cell assay, comprising:
a membrane having a first side and a second, opposing side;
a plurality of pores spanning the membrane from the first side to the second side;
a cell adhesive material layer positioned on the first side of the membrane such that the cell adhesive material layer is exposed through the pores to an environment in contact with the second, opposing side and occludes at least some of the plurality of pores; and
cells positioned on the second side of the membrane and attached to the cell adhesive material layer, wherein each attached cell is associated with a single pore, such that a spatial distribution of the cells on the membrane is directed by a spatial distribution of the pores.