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Systems and methods for identifying and disrupting cellular organelles

Systems and methods for identifying and disrupting cellular organelles description/claims

This application claims priority to U.S. Provisional Application Ser. No. 60/848,513 filed Sep. 29, 2006, the disclosure of which is incorporated herein by reference.

Procedures set forth in this application were supported in part by Grant No. NIH RR 14892 awarded by the National Institute of Health, and by Grant No. AFOSR No. F9620-00-1-0371 awarded by the United States Air Force. The government may have certain rights in any inventions derived therefrom.

Provided herein are systems and methods for the alteration, modification or disruption of target organelles and/or regions in a cell or group of cells.

Systems and methods for disrupting cellular organelles have been developed. Conventional systems generally rely on direct user interface to perform the necessary capture and ablation procedure on a targeted cell. Accordingly, conventional systems rely on the presence of a technician or other end-user in order to design, manipulate, sort, assemble, trap and disrupt a target object, such as a cellular organelle.

The application relates, in general, to the field of systems and methods for altering, modifying or disrupting a target object or portion thereof. A target object includes a cell or selected region in a cell such as an organelle. The cell or portion thereof can be associated with a carrier, such as a bead or other carrier that facilitates movement and/or detection of the cell in an optomechanical system.

In particular, the present application relates to optomechanical systems that correlate image acquisition of a target object with the controlled application of electromagnetic radiation to the target. An optomechanical system provided herein can be used in association with fluidic transport elements that transport and position objects in an environment suitable for imaging and electromagnetic radiation application. Embodiments of a system include a remote controller for remotely controlling image acquisition and radiation application. In some embodiments, a system is adapted to identify a target object in the absence of operator intervention. In other embodiments, a target object is identified by an operator. According to various embodiments, a system can be remotely controlled via a graphical user interface. The user interface may be viewed from a host computer or from a remote site.

In one embodiment, an optomechanical system is provided. The system includes a platform configured to accommodate at least one target object or portion thereof in a fluid medium. A target object includes a cell, an organelle, a cell or organelle complexed with a carrier, or any combination thereof. A system further includes a detector assembly operationally associated with the platform and configured to capture images associated with a target object contained in the fluid medium; at least one radiation source coupled to the detector assembly and operationally configured to emit a pattern of radiation sufficient to specifically disrupt the target object; and a controller operationally associated with the detector assembly and the radiation source. In general, and in the specific embodiments claimed, the controller is configured to coordinate the pattern of radiation emission from the radiation source with the image of the target object captured by the detector assembly. In some aspects the platform is associated with a fluidic flow path. The flow path can be connected to a reservoir that includes a one or more target objects such as cells or portions thereof. In one aspect the fluid flow path is a microfluidic flow path.

In some aspects the object is a cell, such as an embryonic stem cell. In other aspects, the object is an organelle. In still other aspects, a portion of a target object, such as an organelle is targeted for disruption. The portion of a cell can be a nucleus. In other aspects the portion of a cell can be a mitochondrion, chloroplast or any other nucleic acid-containing cellular structure such as a ribosome, nucleolus, inter-chromatin granule cluster, cytoplasmic P-body or any site of replication by a DNA or RNA virus, viroid or other nucleic acid-containing entity.

In some embodiments, a controller associated with a system of the invention can be operated by a user. In some aspects, the user is a remote user.

In other aspects, the controller is configured to determine a profile of the target object based on the images of the target object. The controller can be further configured to synchronize entry of an object in to a flow path associated with a platform. The flow path may be connected to a reservoir. The platform may be associated with image detection by the detector assembly and radiation emission from the radiation source.

In some embodiments, the radiation source included in a system provided herein is a laser. The laser can be a gas laser, a solid-state laser, a tunable dye laser, or semiconductor laser.

In some aspects, the detector assembly includes a complementary metal oxide semiconductor (CMOS) imager, a charge coupled device (CCD) imager, a camera with photosensitive film, a fluorescence imager, a Vidicon camera, or any combination thereof.

In another embodiment, a method of disrupting an object is provided. The method includes directing an object, such as a cell, in to a flow path; imaging a target object or portion thereof; generating a profile of the object based upon the image; and exposing the target object to a radiation emission sufficient to specifically disrupt the target object. In general the pattern of radiation emission is determined by the profile of the object.

In another embodiment, a method for identifying and disrupting a target object is provided. The method includes directing an object in to a flow path; imaging a target object; outlining the image of the object; generating a profile of the object by digitizing the outlined image; transmitting the profile of the object to a controller; contacting the target object or portion thereof to radiation suitable for disrupting the target object or portion thereof, wherein the contacting includes: i) emitting radiation in a pattern determined by the profile of the object; ii) translocating the emitted radiation in a pattern determined by the profile of the object; or iii) maintaining the emitted radiation substantially stationary while translocating the object in a pattern determined by the profile of the object, thereby exposing the target object to a pattern of radiation emission sufficient to specifically disrupt the target object. In some embodiments trapping radiation is applied to the object in order to maintain the position of the object.

• Provisional Patent
• Utility Patent

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