Surgery – Instruments – Light application
Reexamination Certificate
1997-06-04
2002-02-12
Shay, David M. (Department: 3739)
Surgery
Instruments
Light application
C606S001000, C606S010000, C606S013000, C606S017000, C604S019000, C604S020000, C128S898000
Reexamination Certificate
active
06346101
ABSTRACT:
The present invention relates to a new and novel technique for introducing biological materials, such as nucleic acids, proteins and the like, into cells and/or cellular components, such as cell nuclei.
As the scientific community's understanding of the field of molecular biology has grown over the last several years, so has interest in manipulating the biological processes that normally take place within individual cells. An active field of this endeavor is the field of recombinant DNA technology. Recombinant DNA technology typically involves altering the naturally-occurring DNA content of a cell, by addition, subtraction and/or substitution of DNA fragments, so that different proteins are encoded by the cell's DNA.
One well-known technique for introducing exogenous DNA fragments into target cells is called “transformation” and typically involves inserting the exogenous DNA fragments into genetic vehicles, such as plasmids or suitable types of viruses, which are capable of traversing cell membranes. One shortcoming of this technique is that little control can be exerted over which target cells will actually come into contact with the vehicles and, consequently, receive exogenous DNA.
Another well-known technique for introducing exogenous DNA fragments and other biological materials into target cells is known as “microinjection.” Microinjection, which is typically performed under a phase-contrast microscope, typically involves filling a glass microcapillary with the desired materials and injecting the materials into the cytoplasm of a target cell with the air of a micromanipulator and gentle air pressure exerted by a syringe connected to the capillary. Although microinjection overcomes the limitation discussed above in connection with transformation in that tight control can be exerted over the selection of a specific target cell, microinjection suffers from being very time-consuming and requiring a high degree of skill. In addition, vibration, grounding, electrostatic shielding, temperature control, optics, recording equipment, and fabrication of microtools can all be complicating factors.
Another technique that has been suggested in the past has been to coat the tip of a projectile with a biological material to be introduced into a cell and then directing the projectile into the cell.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide a new and novel technique for introducing biological materials into cells and/or cellular components.
It is another object of the present invention to provide a technique as described above which overcomes at least some of the limitations described above in connection with the above-noted existing techniques.
Accordingly, in keeping with the teachings of the present invention, there is disclosed in one embodiment of the present invention a method for introducing desired biological materials into a target cell and/or a target cellular component, the method comprising the steps of (a) providing a light supply, said light supply including an optical fiber having an output end; (b) contacting said output end of said optical fiber with desired biological materials; (c) orienting said output end of said optical fiber towards a target cell and/or a target cellular component; and (d) transmitting a pulse of light of an appropriate energy level through said optical fiber so as to impart momentum to the desired biological materials disposed at said output end and so as to cause ablation of the membrane of the target cell and/or of the target cellular component, whereby the biological materials enter the target cell and/or the target cellular component through the ablated membrane.
The present invention is also directed to a system for carrying out the aforementioned method.
Additional objects, as well as features and advantages, of the present invention will appear from the description to follow. In the description, reference is made to the accompanying drawings which form a part thereof, and in which is shown by way of illustration, specific embodiments for practicing the invention. These embodiments will be described in sufficient detail to enable those skilled in the art to practice the invention, and it is to be understood that other embodiments may be utilized and that structural changes may be made without departing from the scope of the invention. The following detailed description is, therefore, not to be taken in a limiting sense, and the scope of the present invention is best defined by the appended claims.
REFERENCES:
patent: 4309998 (1982-01-01), Aron nee Rosa et al.
patent: 5061265 (1991-10-01), Abela et al.
patent: 5246437 (1993-09-01), Abela
Trokel; “YAG Laser Ophthalmic Microsurgery”; Appleton-Century-Eroft/Norwalk, Ct. pp. 40-41.
Alfano Robert R.
Liu Cheng Hui
Kriegsman & Kriegsman
Research Foundation of City College of New York
Shay David M.
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