Apparatus and method for multiple reactions in small volumes
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
B01L-003/00
C12M-001/00
C12M-001/32
출원번호
US-0811638
(2011-07-25)
등록번호
US-9878328
(2018-01-30)
국제출원번호
PCT/SG2011/000263
(2011-07-25)
§371/§102 date
20130122
(20130122)
국제공개번호
WO2012/011877
(2012-01-26)
발명자
/ 주소
Kim, Namyong
Cheong, Siah Chong
Lye, Melvin Han Wen
Phong, Mark Siew Peng
Li, Li
Seah, Teow Soon
출원인 / 주소
Curiox Biosystems Pte Ltd.
대리인 / 주소
Morgan, Lewis & Bockius LLP
인용정보
피인용 횟수 :
0인용 특허 :
30
초록▼
This describes an apparatus comprising a plate with a number of elements having a hydrophilic surface arranged in an array with an overlay having a hydrophobic surface. A height of the overlay is between about 5% and 100% of the diameter of the elements. The application also discloses a wall circumf
This describes an apparatus comprising a plate with a number of elements having a hydrophilic surface arranged in an array with an overlay having a hydrophobic surface. A height of the overlay is between about 5% and 100% of the diameter of the elements. The application also discloses a wall circumferential to the plate; and a removable grid insertable into the apparatus, said grid comprising dividers enclosing a number of through-holes, said through-holes spaced in the grid to allow alignment of the through-holes of the grid over the elements in the plate when said grid is inserted into the apparatus, said dividers of the grid inserted into the apparatus form sides of wells bottomed by the plate and at least one element on said plate.
대표청구항▼
1. An apparatus for improving retention of non-adherent cells on a plate, the apparatus comprising: the plate comprising a number of elements having a first surface energy arranged in an array with an overlay, on the plate, having a second surface energy, the first surface energy resulting in a hydr
1. An apparatus for improving retention of non-adherent cells on a plate, the apparatus comprising: the plate comprising a number of elements having a first surface energy arranged in an array with an overlay, on the plate, having a second surface energy, the first surface energy resulting in a hydrophilic surface and the second surface energy resulting in a hydrophobic surface, wherein: a diameter of the elements is at least 1 mm; anda height of the overlay having the second surface energy, which results in a hydrophobic surface, is between 5% and 100% of the diameter of the elements;a wall circumferential to the plate; anda removable grid insertable into the apparatus to be positioned over the plate and within the wall of the apparatus, the grid comprising dividers enclosing a number of through-holes, said through-holes spaced in the grid to allow alignment of the through-holes of the grid over the elements in the plate when the grid is inserted into the apparatus, wherein said dividers of the grid inserted into the apparatus form sides of wells bottomed by the plate and at least one element on said plate, the grid has a surface energy that results in a hydrophobic surface, and the grid is distinct and separate from the overlay and the wall. 2. The apparatus of claim 1, wherein the apparatus further comprises a positioning structure adjacent to the interior of the wall so as to guide alignment of the through-holes of the grid over the elements when the grid is inserted into the apparatus. 3. The apparatus of claim 1, wherein the number of through-holes in the grid is the same as the number of elements in the plate. 4. The apparatus of claim 1, wherein the number of though-holes in the grid is smaller than the number of elements in the plate. 5. The apparatus of claim 4, wherein each through-hole is aligned over multiple elements in the plate. 6. The apparatus of claim 1, wherein the grid further comprises baffles on the surface of the grid that touches the plate when the grid is inserted into the apparatus. 7. The apparatus of claim 1, wherein the grid has the second surface energy. 8. The apparatus of claim 1, wherein the grid further comprises at least one structure on the surface of the grid that does not touch the plate when the grid is inserted into the apparatus, said structure sized for a user to grab the grid and insert or remove the grid from the apparatus. 9. The apparatus of claim 1, wherein the grid further comprises a reservoir encompassing at least one through-hole of the grid, said reservoir on the surface of the grid that does not touch the plate when the grid is inserted into the apparatus. 10. The apparatus of claim 1, wherein the dividers of the grid and the wall of the apparatus are of the same height. 11. The apparatus of claim 1, wherein the dividers of the grid are of a height that is higher than the height of the wall of the apparatus. 12. The apparatus of claim 1, wherein the grid further comprises a lid perpendicular to the dividers of the grid attached to the surface of the grid that does not touch the plate when the grid is inserted into the apparatus, wherein the lid covers all of the through-holes of the grid. 13. The apparatus of claim 12, wherein the lid further comprises sides perpendicular to said lid, said sides parallel to and shorter than the dividers of the grid. 14. The apparatus of claim 13, wherein the grid further comprises baffles between the dividers and the lid. 15. The apparatus of claim 1, wherein the height of the overlay over the elements is between 10% and 80% of the diameter of the elements. 16. The apparatus of claim 1, wherein the height of the overlay over the elements is between 20% and 50% of the diameter of the elements. 17. The apparatus of claim 1, wherein the dividers are aligned directly over the hydrophobic surface the overlay having the second surface energy resulting in the hydrophobic surface when the grid is inserted into the apparatus. 18. The apparatus of claim 1, wherein the wall circumferential to the plate defines a reservoir and the apparatus includes an oil within the reservoir. 19. The apparatus of claim 18, wherein the removable grid has a weight that prevents flotation of the removable grid on the oil within the reservoir. 20. The apparatus of claim 1, wherein the diameter of the element is at least 2 mm. 21. The apparatus of claim 1, wherein the overlay is made of polytetrafluoroethylene. 22. A method for performing multiple reactions, comprising: (a) providing (i) an apparatus for improving retention of non-adherent cells on a plate, the apparatus comprising the plate comprising a number of elements having a first surface energy arranged in an array with an overlay, on the plate, having a second surface energy, the first surface energy resulting in a hydrophilic surface and the second surface energy resulting in a hydrophobic surface, wherein a diameter of the elements is at least 1 mm and a height of the overlay having the second surface energy, which results in a hydrophobic surface, is between 5% and 100% of the diameter of the elements; (ii) a wall circumferential to the plate; and (iii) a removable grid insertable into the apparatus to be positioned over the plate and within the wall of the apparatus, the grid comprising dividers enclosing a number of through-holes, said through-holes spaced in the grid to allow alignment of the through-holes of the grid over the elements in the plate when the grid is inserted into the apparatus, wherein said dividers of said grid inserted into the apparatus form sides of wells bottomed by the plate and at least one element on said plate, the grid has a surface energy that results in a hydrophobic surface, and the grid is distinct and separate from the overlay and the wall;(b) adding a first reagent-containing liquid through the through-holes of the grid under conditions where the first reagent in the liquid adheres to the elements on the plate;(c) adding rinsing oil to said plate in an amount necessary to cover said plate;(d) draining excess rinsing oil from the plate resulting from step (c);(e) adding a liquid through the through-holes of the grid, said liquid containing a second reagent suspected of reacting with the first reagent;(f) removing the grid from the apparatus;(g) washing the apparatus; and(h) detecting reaction of the second reagent with the first reagent. 23. The method of claim 22, wherein the first reagent is a cell. 24. The method of claim 23, wherein the cell is selected from the group consisting of an adherent cell, a non-adherent cell, and a permeabilized cell. 25. The method of claim 23, wherein the second reagent is selected from the group consisting of an agonist or an antagonist. 26. The method of claim 22, wherein the first reagent is a binding agent. 27. The method of claim 26, wherein the binding agent is selected from the group consisting of a single-stranded nucleic acid molecule, an antibody, and a ligand. 28. The method of claim 22 further comprising inserting the grid into the apparatus following step (g). 29. The method of claim 22, wherein step (h) is performed using a third reagent. 30. The method of claim 29, wherein the third reagent is detectably labeled.
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