System and method for laser lysis

Abstract

The present invention provides a system and method for lysing individual cells in situ, including the steps of capturing a tissue sample comprising a cellular content, subjecting the tissue sample to a stream of continuous fluid flow, lysing a selected area of the tissue sample with a laser, thereby releasing at least a portion of the cellular content from the tissue sample, recovering at least one target molecule from the cellular content in the stream, and processing the at least one target molecule.

We claim: 
     
         1 . A method of lysing individual cells in situ, comprising:
 lysing a selected area of a tissue sample being subjected to a steam of continuous fluid flow with a laser, thereby releasing at least a portion of a cellular content from the tissue sample.   
     
     
         2 . The method of  claim 1 , wherein the tissue sample is a live tissue, and wherein the selected area is a single cell of the live tissue. 
     
     
         3 . The method of  claim 1 , further comprising recovering at least one target molecule from cellular content in the stream. 
     
     
         4 . The method of  claim 3 , further comprising processing the at least one target molecule. 
     
     
         5 . The method of  claim 4 , wherein processing comprises single-cell quantitative in situ RT-PCR. 
     
     
         6 . The method of  claim 1 , wherein the tissue sample has a maximum dimension of about 200 μm to about 300 μm. 
     
     
         7 . The method of  claim 1 , wherein the laser is a two-photon laser. 
     
     
         8 . The method of  claim 1 , wherein the step of capturing further includes collecting the tissue sample with a tissue collection device. 
     
     
         9 . The method of  claim 8 , further including aspirating the tissue sample with the tissue collection device, and automatically depositing the tissue sample on an analysis platform. 
     
     
         10 . The method of  claim 9 , wherein the analysis platform is removably coupled to the tissue collection device. 
     
     
         11 . The method of  claim 9 , wherein the analysis platform is a microfluidic chip comprising a cage configured to retain the tissue sample. 
     
     
         12 . The method of  claim 11 , wherein the cage comprises a fluid channel and a plurality of posts spaced apart within the fluid channel. 
     
     
         13 . A system for lysing cells, comprising:
 a microfluidic chip having a fluid channel and a cage disposed within the fluid channel, the cage sized to capture a tissue sample;   a microscope for observing the tissue sample;   a laser for irradiating a selected area of the tissue sample; and   a downstream module coupled to the microfluidic chip for processing a target molecule collected from the tissue sample;   wherein irradiating the tissue sample with the laser lyses the selected area of the tissue sample, thereby releasing the target molecule from the tissue sample into the fluid channel.   
     
     
         14 . The system of  claim 13 , wherein the laser is a two-photon laser. 
     
     
         15 . The system of  claim 13 , wherein the module is a quantitative RT-PCR module. 
     
     
         16 . A device, comprising:
 an apparatus including a body defining a passage, a capillary in fluid communication with an inlet of the passage, and an output port in fluid communication with an outlet of the passage; and   a microfluidic chip removably coupled to the body, the microfluidic chip including a first fluid channel having an inlet and an outlet in communication with the passage, and a cage positioned between the inlet and the outlet, the cage comprising a plurality of structures sized to retain a tissue sample in the first fluid channel.   
     
     
         17 . The device of  claim 16 , further comprising an RT-PCR module in communication with the first fluid channel. 
     
     
         18 . The device of  claim 16 , wherein the microfluidic chip further comprises a second fluid channel in communication with the first fluid channel downstream of the cage, and a third fluid channel in communication with the first fluid channel downstream of the second fluid channel,
 wherein the second fluid channel is in communication with a source of RT-PCR reagents, and wherein the third fluid channel is in communication with a source of material for encapsulating the contents of the first fluid stream and the second fluid stream.   
     
     
         19 . The device of  claim 18 , further comprising a motorized platform for dispensing liquid from the microfluidic chip into a container. 
     
     
         20 . The device of  claim 19 , further comprising a sensor for monitoring at least one of the motorized platform and the container, wherein the sensor is configured to reduce carryover contamination.