US5795755A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method of implanting living cells by laser poration at selected sites
Assignee: Individual
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Filing: 1996-02-22
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Granted: 1998-08-18
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Expires: 2016-02-22
Abstract
Abstract A method and apparatus for operating on living cells in order to transfer material to the interior of selectively targeted cells is disclosed. The operation may take place in a reaction chamber in which a group of cells are disposed, only some of which are desired to receive material which serves to transform or otherwise affect the cells. In one embodiment, certain of the cells in the reaction chamber are targeted by computerized image analysis of images of the cells as generated by confocal microscopy. The targeted cells are then transfected with genetic material using a computer controlled laser beam. The laser beam creates pores in the membranes of the targeted cells through which genetic material also present within the reaction chamber may pass. An electric field may also be employed to facilitate pore formation.
Inventors
- 👤 Jerome H. Lemelson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 17:38
CPC Classifications
C; C12; C12M; C12M35/00; C12M35/02
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
140 words
A method of transferring exogenous material into living cells comprising the steps of: disposing a specimen of said living cells within a reaction chamber together with the exogenous material; sensing a plurality of images of the surfaces of said living cells and sections of said living cells, and generating cell sensing signals from said plurality of images; transmitting said cell sensing signals to a computer and operating said computer to identify desired cells in the specimen, identify desired specific locations on the surfaces of said desired cells, and determine location coordinates within the reaction chamber for the desired specific locations; and under control of the computer, generating and directing at least one source of laser light energy at the location coordinates, whereby said laser light energy creates transient pores in the desired specific locations through which the exogenous material passes.
Claim 2
Independent Claim
140 words
A method of transferring exogenous material into living cells comprising the steps of: disposing a specimen of said living cells within a reaction chamber together with the exogenous material; sensing a plurality of images of the surfaces of said living cells and sections of said living cells, and generating cell sensing signals from said plurality of images; transmitting said cell sensing signals to a computer and operating said computer to identify desired cells in the specimen, identify desired specific locations on the surfaces of said desired cells, and determine location coordinates within the reaction chamber for the desired specific locations; and under control of the computer, generating and directing at least one source of laser light energy at the location coordinates, whereby said laser light energy creates transient pores in the desired specific locations through which the exogenous material passes.
Claim 3
Dependent Claim
14 words
The method of claim 1, wherein the exogenous material comprises fragments of foreign DNA.
Claim 4
Dependent Claim
20 words
The method of claim 1, wherein the exogenous material comprises monoclonal antibodies directed against intracellular targets within the desired cells.
Claim 5
Dependent Claim
23 words
The method of claim 1, wherein said plurality of images are sensed in the spectrum of radiation fluorescently emitted from said living cells.
Claim 6
Dependent Claim
28 words
The method of claim 1, further comprising applying an electric field to the specimen at a low intensity just sufficient to facilitate the creation of said transient pores.
Claim 7
Dependent Claim
20 words
The method of claim 1, wherein said plurality of images are sensed by generating confocal microscopic images of the specimen.
Claim 8
Dependent Claim
26 words
The method of claim 6, wherein said confocal microscopic images are generated by scanning the living cells with light passing through a plurality of moving apertures.
Claim 9
Independent Claim
184 words
An apparatus for transferring exogenous material into living cells comprising: a chamber defining a volume of space sufficient to retain and hold a specimen of said living cells and said exogenous material within its boundaries and having a means for accessing the inside of said chamber and at least one transparent window; a confocal microscope for sensing a plurality of images of the surfaces of said living cells and sections of said living cells; a photo detector array for generating cell sensing signals from said plurality of images; a computer for analyzing said cell sensing signals in order to identify desired cells in the specimen and for determining location coordinates within the reaction chamber for the desired cells; a pair of electrodes for applying an electric field to the specimen under control of said computer in order to facilitate formation of pores in the membranes of the desired cells; and a radiant energy generating device under control of said computer for directing radiant energy at the desired cells in order to create the pores in the membranes thereof through which said material may pass.
Claim 10
Independent Claim
184 words
An apparatus for transferring exogenous material into living cells comprising: a chamber defining a volume of space sufficient to retain and hold a specimen of said living cells and said exogenous material within its boundaries and having a means for accessing the inside of said chamber and at least one transparent window; a confocal microscope for sensing a plurality of images of the surfaces of said living cells and sections of said living cells; a photo detector array for generating cell sensing signals from said plurality of images; a computer for analyzing said cell sensing signals in order to identify desired cells in the specimen and for determining location coordinates within the reaction chamber for the desired cells; a pair of electrodes for applying an electric field to the specimen under control of said computer in order to facilitate formation of pores in the membranes of the desired cells; and a radiant energy generating device under control of said computer for directing radiant energy at the desired cells in order to create the pores in the membranes thereof through which said material may pass.
Claim 11
Dependent Claim
28 words
The apparatus of claim 8, wherein said confocal microscope contains a plurality of moving apertures through which illuminating and reflected light passes in order to scan the specimen.
Claim 12
Independent Claim
199 words
A method of selectively transferring desired exogenous material into living cells comprising the steps of: disposing a specimen of said living cells within a reaction chamber together with the exogenous material; sensing a plurality of images of the surfaces of said living cells and sections of said living cells, and generating cell sensing signals and exogenous material sensing signals from said plurality of images; transmitting said cell sensing signals and said material sensing signals to a computer and operating said computer to identify desired cells in the specimen, identify desired specific locations on the surfaces of the desired cells, identify specific locations of desired exogenous material, and determine location coordinates within the reaction chamber for poration sites, which poration sites are said desired specific locations on the surfaces of the desired cells which are sufficiently close to said specific locations of desired exogenous material to permit the selective passage of said desired exogenous material into the desired cells; and under control of the computer, generating and directing at least one source of laser light energy at the location coordinates, whereby said laser light energy creates transient pores in the poration sites through which the desired exogenous material selectively passes.
Claim 13
Independent Claim
199 words
A method of selectively transferring desired exogenous material into living cells comprising the steps of: disposing a specimen of said living cells within a reaction chamber together with the exogenous material; sensing a plurality of images of the surfaces of said living cells and sections of said living cells, and generating cell sensing signals and exogenous material sensing signals from said plurality of images; transmitting said cell sensing signals and said material sensing signals to a computer and operating said computer to identify desired cells in the specimen, identify desired specific locations on the surfaces of the desired cells, identify specific locations of desired exogenous material, and determine location coordinates within the reaction chamber for poration sites, which poration sites are said desired specific locations on the surfaces of the desired cells which are sufficiently close to said specific locations of desired exogenous material to permit the selective passage of said desired exogenous material into the desired cells; and under control of the computer, generating and directing at least one source of laser light energy at the location coordinates, whereby said laser light energy creates transient pores in the poration sites through which the desired exogenous material selectively passes.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-08-17 | 2002-02-16 | 2002-08-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-08-17 | 2006-02-16 | 2006-08-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-08-17 | 2010-02-16 | 2010-08-18 | $7700.00 | Window Closed / Lapsed |