Microelectromechanical devices useful for manipulating cells or embryos, kits thereof, methods of making same, and methods of use thereof

Abstract

The present invention relates generally to microelectromechanical systems (MEMS) devices for the manipulation of cells or groups of cells, such as oocytes, embryos, and sperm. In particular, the present invention relates to Cell Labeling MEMS devices ( 2 F), Microinjection MEMS devices, IntraCytoplasmic Sperm Injection (“ICSI”) MEMS devices, Zona Coring MEMS devices, Enucleation MEMS devices, Enucleation/Nuclear Transfer MEMS devices, and Cytoplasmic Transfer MEMS devices. The present invention also relates to kits containing the MEMS devices of the present invention.

1 - 47 . (canceled) 
   
   
       48 . A microinjection MEMS device for injecting a fluid, a suspension or a material into a cell or group of cells comprising:
 (a) a first substrate comprising at least one well for holding the cell or group of cells and wherein the well comprises at least one hollow protuberance for penetrating the cell or group of cells and wherein the well is in fluid communication with a fluid transfer channel wherein the fluid transfer channels permits the fluid to enter the hollow protuberance and to then enter the cell; and   (b) a second substrate comprising an input manifold in fluid communication with the fluid transfer channel wherein the input manifold allows for the input of the fluid, suspension or material into the hollow protuberances.   
   
   
       49 . The microinjection MEMS device of  claim 48  further comprising a pumping means. 
   
   
       50 . The microinjection MEMS device of  claim 48  wherein the well is cube-shaped. 
   
   
       51 . The microinjection MEMS device of  claim 50  wherein the cube-shaped well is from about 50 μm to about 200 μm in length per side. 
   
   
       52 . The microinjection MEMS device of  claim 48  wherein the well is conical-shaped. 
   
   
       53 . The microinjection MEMS device of  claim 48  wherein the hollow protuberance is a needle. 
   
   
       54 . The microinjection MEMS device of  claim 48  wherein the hollow protuberance is from about 0.01 μm to about 100 m in diameter. 
   
   
       55 . The microinjection MEMS device of  claim 48  further comprising a coating on the surfaces of the device exposed to the cells. 
   
   
       56 . The microinjection MEMS device of  claim 55  wherein the coating is a polypeptide, peptide or protein. 
   
   
       57 . The microinjection MEMS device of  claim 56  wherein the polypeptide is polylysine. 
   
   
       58 . The microinjection MEMS device of  claim 48  wherein the hollow protuberance acts as an emitter to facilitate piercing the cell or group of cells. 
   
   
       59 . A method of making a microinjection MEMS device comprising the steps of:
 (a) etching a plurality of parallel channels on a first side of a plurality of silicon wafers in which the wafers each have a second unetched side;   (b) silicon fusion bonding the unetched side of a plurality of silicon wafers of step (a) to the etched side of a plurality of silicon wafers of step (a) such that the etched channels are in parallel to form a mega-laminate wherein the mega-laminate has a plurality of channels;   (c) cutting the mega-laminate at an angle perpendicular to the long axis of the etched channels thereby forming a slice of the mega-laminate having a top surface and a bottom surface wherein each surface exposes an end of the channel;   (d) silicon fusion bonding the bottom surface of the slice of the mega-laminate to the etched side of a channel-etched base-plate wafer;   (e) depositing a first mask on the top surface of the slice of the mega-laminate such that a region surrounding each channel end is free of mask;   (f) etching the mask to form a plurality of wells;   (g) depositing a second mask on the mega-laminate top surface such that a boarder forms around each channel end such that material around the channel is not etched; and   (h) etching the second mask thereby forming a plurality of hollow protuberances.   
   
   
       60 . The method of making a microinjection MEMS device of  claim 59  further comprising applying a coating to the mega-laminate top surface after step (h). 
   
   
       61 . The method of making a microinjection MEMS device of  claim 60  wherein the coating is a polypeptide, peptide or protein. 
   
   
       62 . The method of making a microinjection MEMS device of  claim 61  wherein the polypeptide is polylysine. 
   
   
       63 . A microinjection MEMS device kit for injecting a fluid, a suspension or a material into a cell or group of cells comprising:
 (a) a centrifugal platter for applying a centripetal force to a cell or group of cells contained within a MEMS device wherein the centrifugal platter comprises a circular disk, a plurality of ports for holding the MEMS devices and a securing means to secure the platter to a spinner or driving means; and   (b) at least one microinjection MEMS device of  claim 48 .   
   
   
       64 . The microinjection MEMS device kit of  claim 63  wherein the microinjection MEMS device is permanently affixed to the centrifugal platter. 
   
   
       65 . The microinjection MEMS device kit of  claim 64  wherein
 (a) the centrifugal platter comprises a plurality of grooves arranged in a concentric pattern and wherein each groove has an inner and outer edge;   (b) at least one microinjection MEMS device is bonded to the outer edge of a groove in an orientation such that the axis of each well of the microinjection MEMS device is horizontal to the plane of the centrifugal platter; and   (c) the inner edge of the grooves forming divided compartments comprising a single well which restrict the movement of materials from one compartment containing a containing a single well to another compartment.   
   
   
       66 . A method of using a microinjection MEMS device kit of  claim 63  comprising the steps of:
 (a) filling the input manifold of at least one microinjection MEMS device resident on a centrifugal platter with a fluid;   (b) loading the fluid-filled wells of step (b) with at least one oocyte or embryo;   (c) placing the microinjection MEMS/centrifugal platter into a centrifuge; and   (d) applying a centripetal force on the microinjection MEMS device kit by rotating the kit using a spinner or driving means.   
   
   
       67 . A method of using a microinjection MEMS device kit of  claim 63  comprising the steps of:
 (a) filling the grooves of the centrifugal platter with a fluid;   (b) loading the grooves of the centrifugal platter with at least one oocyte or embryo; and   (c) applying a centripetal force to the kit whereby the oocyte or embryo makes contact with the hollow protuberance of the microinjection MEMS device and the hollow protuberance penetrates the surface of the oocyte or embryo.   
   
   
       68 . A method of using a microinjection MEMS device kit of  claim 63  wherein, upon rotation of centrifugal platter, a volume of fluid is caused to enter the oocyte or embryo in the cell well through the hollow protuberance. 
   
   
       69 . A microinjection MEMS array for injection of a fluid, a suspension or a material into a cell or group of cells comprising:
 (a) a first substrate comprising at least one well for accepting a cell or group of cells and wherein the well comprises a hollow protuberance for penetrating the cell or group of cells;   (b) a second substrate comprising a fluid handling means in fluid communication with said hollow protuberance; and   (c) a central loading manifold for loading a fluid into the array.   
   
   
       70 . A microinjection MEMS array of  claim 69  wherein the fluid handling means is a dynamic hydropressure column. 
   
   
       71 . A microinjection MEMS array of  claim 69  wherein the device is embedded in a centrifugal platter. 
   
   
       72 . A method of using the microinjection MEMS array of  claim 69  comprising:
 (a) applying an inertial force to the device using a centripetal (angular) acceleration means brought about by rotation of the centrifugal platter.   
   
   
       73 . A microinjection MEMS array for injecting a fluid, a suspension or a material into a cell or group of cells comprising:
 (a) a central loading manifold for loading the fluid, suspension or material into the array;   (b) a plurality of wells for receiving cells;   (c) a hollow protuberance within each well for penetrating the cell and injecting the fluid, suspension or material; and   (d) a plurality of dynamic hydropressure columns in fluid communication with the central loading manifold and with the hollow protuberances wherein the dynamic hydropressure columns provide pressure for forcing the fluid, suspension or material through the hollow protuberance and into the cell.   
   
   
       74 . The microinjection MEMS array of  claim 73  further comprising at least one valve in the dynamic hydropressure column for modulating fluid flow. 
   
   
       75 . A microinjection MEMS array of  claim 74  wherein each valve is in operable communication with a controller to control the fluid, suspension or material flowing into the cell. 
   
   
       76 . A microinjection MEMS device of  claim 75  wherein the operable communication is mediated by electronic circuits. 
   
   
       77 . A microinjection MEMS device of  claim 75  wherein the operable communication is mediated by Electro-optical circuits. 
   
   
       78 . A method of using a microinjection MEMS device of  claim 73  comprising:
 (a) loading at least one cell or group of cells into an injection domain of the microinjection MEMS device; and   (b) applying a centripetal force to the microinjection MEMS device thereby causing penetration of the cell or group of cells by the hollow protuberance of the microinjection MEMS device; and deposition of a substance in the cell or group of cells from the hollow protuberance.   
   
   
       79 . A method of using a microinjection MEMS device of  claim 73  comprising:
 (a) loading at least one cell or group of cells into the well of the microinjection MEMS array; and   (b) rotating the microinjection MEMS device whereby the cell or group of cells is thrust upon the hollow protuberance resident within well and simultaneously providing for the passive movement of fluid through dynamic hydropressure columns provides pressure to push fluid into cell. (c)   
   
   
       80 . The method of using the microinjection MEMS of  claim 79  further comprising a valve providing for variable fluid flow from the dynamic hydropressure column into the cell when activated by a controller by way of a circuit. 
   
   
       81 - 134 . (canceled)