Method and device for selectively targeting cells within a...

Image analysis – Applications – Biomedical applications

Reexamination Certificate

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C359S368000

Reexamination Certificate

active

06804385

ABSTRACT:

BACKGROUND OF THE INVENTION
This invention relates to methods and devices for selectively identifying and individually manipulating particles in mixtures and more specifically to methods and devices for selectively removing unwanted target cells from a tissue with an energy beam.
Previous methods of removing target cells from tissues have been developed based upon the ability to effectively separate target cells from the tissue or availability of toxic chemical agents that are delivered specifically to the target cells. Effective separation of target cells is difficult to achieve due to the difficult balance to be struck between providing conditions that are rigorous enough to remove target cells from their native tissues and gentle enough not to damage other cells desired to be maintained in the tissue. Furthermore, many of the separation methods require extensive disruption of the tissue thereby precluding or rendering difficult reconstitution of viable tissue with the remaining cells. Although, toxic agents can be delivered to the target cells while still in the tissue, the degree of specificity required to prevent collateral damage to surrounding tissue is often difficult to achieve.
Thus, there is a need for apparatus and methods for rapidly and efficiently identifying and targeting particular cells within complex populations found in biological tissues. The present invention satisfies this need and provides related advantages as well.
SUMMARY OF THE INVENTION
The invention provides an apparatus for electromagnetically affecting a particle of interest in a specimen. The apparatus includes (a) a stage capable of supporting the specimen; (b) a detector including at least one camera, wherein the detector is capable of resolving a particle of interest within the specimen; (c) a means for locating the particle of interest in three dimensions; (d) a means for focusing electromagnetic radiation to a focal volume within the specimen; and (e) a means for adjusting the relative positions of the stage and electromagnetic radiation focusing means, thereby positioning the particle of interest within the focal volume.
The invention further provides a method for electromagnetically affecting a particle of interest in a specimen. The method includes the steps of (a) obtaining a plurality of nonidentical two-dimensional sectional representations of the specimen in which the particle of interest is discernable in at least one of the sectional representations; (b) combining the plurality of sectional two-dimensional representations to produce a three-dimensional representation of the specimen; (c) locating the particle of interest in three dimensions based on the three-dimensional representation; and (d) focusing electromagnetic radiation to a focal volume, the focal volume intersecting a portion of the specimen containing the particle of interest, wherein the radiation within the focal volume substantially affects only the portion of the specimen within the focal volume, the portion of the specimen within the focal volume being surrounded by an envelope of substantially unaffected specimen.
Further provided is a method for electromagnetically affecting a particle of interest in a specimen. The method includes the steps of (a) focusing a plurality of detectors on a plurality of focal planar regions in a specimen; (b) obtaining a plurality of two-dimensional sectional representations each corresponding to one of the focal planar regions, wherein a particle of interest can be discerned in at least one of the two-dimensional sectional representations; (c) storing the plurality of two-dimensional sectional representations in a computer memory; (d) combining the plurality of two-dimensional sectional representations to produce a three-dimensional representation of at least a portion of the specimen; (e) locating the particle of interest in the specimen based on the three-dimensional representation of at least a portion of the specimen; and (f) focusing electromagnetic radiation to a focal volume, the focal volume intersecting a portion of the specimen containing the particle of interest, wherein the radiation within the focal volume substantially affects only the portion of the specimen within the focal volume, the portion of the specimen within the focal volume being surrounded by an envelope of substantially unaffected specimen.
The invention also provides a method for electromagnetically affecting a particle of interest in a specimen. The method includes the steps of: (a) obtaining a plurality of two-dimensional sectional representations of the specimen at different Z-levels, wherein the particle of interest is discernable in at least one of the sectional representations; (b) selecting a two-dimensional sectional representation from the plurality of two-dimensional sectional representations wherein the particle of interest is in focus; (c) locating the particle of interest in the specimen with reference to X and Y coordinates of the particle in the selected two-dimensional sectional representation and the Z-level of the selected two-dimensional sectional representation; and (d) focusing electromagnetic radiation to a focal volume, the focal volume intersecting a portion of the specimen containing the particle of interest, wherein the radiation within the focal volume substantially affects only the portion of the specimen within the focal volume, the portion of the specimen within the focal volume being surrounded by an envelope of substantially unaffected specimen.


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