Systems and methods for identifying and disrupting cellular organelles

Abstract

This invention relates to optomechanical systems and methods for altering, modifying or disrupting a target object. Such systems and methods are used for, for example, ablating the endogenous nucleus in a cell.

1 . An optomechanical system comprising:
 a) a platform configured to accommodate at least one target object in a medium;   b) a detector assembly functionally associated with the platform and configured to capture images of the target object or a portion thereof;   c) at least one emitter functionally associated with the platform and configured to emit patterns of radiation suitable for:
 i) optically trapping the target object; and 
 ii) disrupting the target object or portion thereof; 
   d) a controller operatively associated with the detector assembly and the emitter, wherein the controller is configured to coordinate the pattern of radiation emission from the emitter with the image of the target object, or portion thereof, captured by the detector assembly.   
     
     
         2 . The system of  claim 1 , wherein the target object is selected from the group consisting of a cell, an organelle, a cell or organelle complexed with a carrier, or any combination thereof. 
     
     
         3 . The system of  claim 1 , wherein the platform is associated with a flow path fluidly connected with a reservoir comprising one or more target objects. 
     
     
         4 . The system of  claim 1 , wherein the flow path is a microfluidic flow path. 
     
     
         5 . The system of  claim 2 , wherein the cell is a stem cell. 
     
     
         6 . The system of  claim 5 , wherein the stem cell is an embryonic stem cell. 
     
     
         7 . The system of  claim 2 , wherein the organelle is a nucleus, a mitochondria, a protoplast, a chloroplast or a centrosome. 
     
     
         8 . The system of  claim 1 , wherein the controller is operated by a user. 
     
     
         9 . The system of  claim 8 , wherein the user is a remote user. 
     
     
         10 . The system of  claim 1 , wherein the controller is configured to determine a profile of the target object based on the images of the target object. 
     
     
         11 . The system of  claim 3 , wherein the controller is further configured to synchronize entry of a target object in to the flow path from the reservoir with image detection by the detector assembly and radiation emission from the emitter. 
     
     
         12 . The system of  claim 1 , wherein the emitter is a laser. 
     
     
         13 . The system of  claim 12 , wherein the laser is a gas laser, a solid-state laser, a tunable dye laser, or semiconductor laser. 
     
     
         14 . The system of  claim 1 , wherein the detector assembly comprises 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. 
     
     
         15 . A method comprising:
 a) directing a target object or portion thereof to a platform;   b) imaging the target object or portion thereof;   c) generating a profile of the target object or portion thereof based upon the image; and   d) contacting the target object or portion thereof to radiation suitable for disrupting the target object or portion thereof, wherein the contacting comprises:
 i) emitting the 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. 
   
     
     
         16 . The method of  claim 15 , further comprising contacting the target object or portion thereof with trapping radiation. 
     
     
         17 . The system of  claim 15 , wherein the target object is selected from the group consisting of a cell, an organelle, a cell or organelle complexed with a carrier, or any combination thereof. 
     
     
         18 . The method of  claim 17 , wherein the object is a cell. 
     
     
         19 . The method of  claim 18 , wherein the cell is an embryonic stem cell. 
     
     
         20 . The method of  claim 17 , wherein the organelle is a nucleus. 
     
     
         21 . The method of  claim 15 , wherein the radiation is emitted by a laser. 
     
     
         22 . The method of  claim 21 , wherein the laser is a gas laser, a solid-state laser, a tunable dye laser, or semiconductor laser. 
     
     
         23 . A method for identifying and disrupting a target object or portion thereof, the method comprising:
 a) directing a target object in to a platform;   b) imaging a target object or portion thereof;   c) outlining the image of the target object or portion thereof;   d) generating a profile of the target object or portion thereof by digitizing the outlined image;   e) transmitting the profile of the target object or portion thereof to an electronic controller operatively associated with an emitter;   f) contacting the target object or portion thereof with radiation emission suitable for:
 i) optically trapping the target object or portion thereof; and 
 ii) disrupting the target object or portion thereof. 
   
     
     
         24 . The method of  claim 23 , wherein contacting the target object or portion thereof for disrupting the object or portion thereof comprises:
 a) emitting the radiation in a pattern determined by the profile of the object;   b) translocating the emitted radiation in a pattern determined by the profile of the object; or   c) maintaining the emitted radiation substantially stationary while translocating the object in a pattern determined by the profile of the object.   
     
     
         25 . The method of  claim 22 , wherein translocating the emitted radiation comprises scanning by a scanning mirror.