Method and apparatus for disrupting cells

Claim 2

The method of claim 2 in which said natural frequency of oscillation is at least 8,000 c.p.s.

Claim 3

The method of claim 1 in which said mass of cells are intermittently discharged by a controlled frequency of oscillation.

Claim 4

The method of claim 4 in which said controlled frequency of oscillation is at least 15 c.p.s.

Claim 5

The method of claim 1 in which said mass of cells is comprised of plant cells.

Claim 6

The method of claim 6 wherein said plant cells are in the form of a viscous paste.

Claim 7

The method of claim 1 in which said mass of cells is comprised of animal cells.

Claim 8

The method of claim 8 wherein said animal cells are in the form of a viscous paste.

Claim 9

The method of claim 1 including the step of: maintaining said relatively high pressure on said mass of cells prior to discharge thereof.

Claim 10

The method of claim 10 in which: said mass of cells are maintained at a pressure of at least 10, 000 p.s.i., and said relatively low pressure environment is at atmospheric pressure.

Claim 11

The method of claim 10 wherein said mass of cells are maintained at a relatively high pressure for a period of 1 second to 30 minutes, prior to initiating the intermittent discharge thereof.

Claim 12

The method of claim 10 including the step of: heat-sinking said mass of cells in order to prevent denaturation caused by excessive heating of said cell mass.

Claim 13

The method of claim 1 wherein said portion of said mass of cells constitutes a thin-film of essentially monocellular thickness, in order to eliminate the possibility of buffering in the vertical plane, thereby ensuring the exposure of each discharged cell to rapid decompression and disruption.

Claim 14

The method of claim 1 wherein said mass of cells is intermittently discharged through a dynamic orifice which is oscillated such that the height of said dynamic orifice varies from zero to a height of less than 4 microns.

Claim 15

Apparatus for disrupting the cellular structure of cells comprising: means for confining a mass of cells under relatively high pressure; means for maintaining a substantially uniform relatively high pressure on said mass of cells to be disrupted; dynamic orifice means cooperating with said confining means, said dynamic orifice means being in communication with a relatively low pressure environment, said dynamic orifice means being operable to oscillate at a frequency which is sufficiently high that a portion of said mass of cells under pressure is intermittently discharged into said relatively low pressure environment, the pressure of said environment being sufficiently low that said discharged cells are disrupted due to rapid decompression caused by the exposure of said discharged cells to said relatively low pressure environment; and means for collecting said disrupted cells for subsequent processing.

Claim 16

The apparatus of claim 16 in which said dynamic orifice means comprises: an oscillating mEmber, said member being adapted to oscillate at a frequency which is sufficiently high that a portion of said mass of cells under pressure is intermittently discharged; and means cooperating with said oscillating member, for the purpose of periodically opening and closing said dynamic orifice means, in order to intermittently discharge said mass of cells to said relatively low pressure environment.

Claim 17

Apparatus for disrupting the cellular structure of cells comprising: means for confining a mass of cells under relatively high pressure; means for maintaining said relatively high pressure on said mass of cells to be discharged; dynamic orifice means cooperating with said confining means, said dynamic orifice means including an oscillating annular member, said member being adapted to oscillate at a frequency which is sufficiently high that a portion of said mass of cells under pressure is intermittently discharged and means cooperating with said oscillating member for the purpose of periodically opening and closing said orifice means in order to intermittently discharge said mass of cells to said relatively low pressure environment and said dynamic orifice means being in communication with a relatively low pressure environment, said dynamic orifice means being operable to oscillate at a frequency which is sufficiently high that a portion of said mass of cells under pressure is intermittently discharged into said relatively low pressure environment, the pressure of said environment being sufficiently low that said discharged cells are disrupted due to rapid decompression caused by the exposure of said discharged cells to said relatively low pressure environment; and means for collecting said disrupted cells for subsequent processing.

Claim 18

The apparatus of claim 18 in which said cooperating means includes: a tapered seat means for said annular oscillating member; and flexible sealing means positioned within said seat means, said sealing means contacting said annular oscillating member when said dynamic orifice means is in the closed position.

Claim 19

The apparatus of claim 19 in which said flexible sealing means comprises a ring assembly including: a resilient O-ring positioned within a concentric support ring, said support ring having an inner diameter which is greater than the outer diameter of said resilient O-ring, said support ring possessing a plurality of notches spaced along the outer periphery thereof, in order to prevent said O-ring from locking said ring assembly when torques or prepressures are applied to said ring assembly.

Claim 20

The apparatus of claim 20 wherein said annular oscillating member comprises a polished disc.

Claim 21

The apparatus of claim 21 wherein said dynamic orifice means communicates with said relatively low pressure environment by means of a plurality of radial slots which are formed at spaced intervals in the lower surface of a disc nut, said disc nut being threadedly engaged within a housing member and positioned adjacent to said polished disc of said apparatus for the purpose of securing said polished disc within said housing member.

Claim 22

The apparatus of claim 21 in which said polished disc and said resilient O-ring is adapted to function as a heat sink in order to prevent denaturation of said mass of cells caused by excessive heating of the cells.

Claim 23

The apparatus of claim 18 in which said annular oscillating member is adapted to oscillate at a natural frequency which is determined by the dimensions of said annular oscillating member and said cooperating means.

Claim 24

The apparatus of claim 24 wherein said natural frequency is at least 8000 c.p.s.

Claim 25

The apparatus of claim 18 in which said annular oscillating member is adapted to oscillate at a controlled frequency.

Claim 26

The apparatus of claim 26 wherein said controlled frequency is at least 15 c.p.s.

Claim 27

The apparatus of claim 16 in which: said maintaining means maintains said mass of cells at a pressure of at leasT 10,000 p.s.i., and said low pressure environment is at atmospheric pressure.

Claim 28

The apparatus of claim 17 in which said oscillating member is adapted to oscillate such that said mass of cells are intermittently discharged in a radial direction around the outer periphery of said oscillating member.

Claim 29

The apparatus of claim 17 in which the mode of oscillation of said oscillating member is such that only a thin film of said cell mass is intermittently discharged, said thin film being essentially of monocellular thickness, thus precluding the possibility of buffering in the vertical plane, in order to ensure that each discharged cell is exposed to rapid decompression and disruption.

Claim 30

The apparatus of claim 21 in which said polished disc is adapted to undergo a compound mode of oscillation, said compound mode including an orbital component and a vertical component, thereby generating a dynamic orifice of revolution, as well as a dynamic orifice which is variable in rectilinear dimensions.

Claim 31

The apparatus of claim 18 in which the height of said dynamic orifice means varies from zero to a height of less than 4 microns, and the circumference of said orifice means is substantially 11 inches.

Claim 32

Apparatus for disrupting the cellular structure of cells comprising: means defining an open-ended chamber for confining a mass of cells under relatively high pressure; means for maintaining said relatively high pressure on said mass of cells to be disrupted; dynamic orifice means disposed at the open end of said chamber for providing at least a single dynamic orifice between said chamber and a relatively low pressure environment, said orifice means being adapted to oscillate at a frequency which is sufficiently high so that said dynamic orifice is intermittently substantially closed and opened to a height not exceeding roughly 4 microns so that a portion of said mass of cells under pressure is intermittently discharged into said relatively low pressure environment, while said orifice is open, the pressure of said environment being sufficiently low that said discharged cells are disrupted due to rapid decompression caused by the exposure of said discharged cells to said relatively low pressure environment; and means for collecting said disrupted cells for subsequent processing.

Claim 33

Apparatus as in claim 33 wherein said orifice means is adapted to oscillate so that at least some sectors of said orifice do not have the same height at the same time.