Methods and systems for using encapsulated microbubbles to process biological samples

Abstract

Methods and systems for using encapsulated microbubbles to process biological samples are disclosed. According to one aspect, a method for using encapsulated microbubbles to process a biological sample includes creating a mixture comprising encapsulated microbubbles mixed with a biological sample and adding activation energy to the mixture to cause at least some of the microbubbles to oscillate or burst and thereby process the sample, including effecting cell lysis, shearing DNA, and/or performing tissue dispersion.

1 . A system for delivering microbubbles to a biological sample, the system comprising:
 a container for holding a solution of microbubbles or a solution that will formulate encapsulated microbubbles when processed; and   at least one outlet for delivering the solution of microbubbles to a biological sample.   
     
     
         2 . The system of  claim 1  comprising an energy source for providing activation energy to the microbubbles that causes the microbubbles to oscillate or burst and thereby process the sample. 
     
     
         3 . The system of  claim 2  wherein the activation energy comprises thermal, sonic, or light energy, optionally wherein the activation energy is ultrasound having a frequency in the range from 0.01 MHz to 10 MHz. 
     
     
         4 . (canceled) 
     
     
         5 . The system of  claim 2  wherein the biological sample comprises DNA or DNA that has been cross-linked to protein and wherein processing the sample comprises shearing the DNA. 
     
     
         6 . The system of  claim 2  wherein the biological sample comprises cells and wherein processing the sample comprises effecting cell lysis, optionally wherein the cells comprise bacteria or yeast cells. 
     
     
         7 . (canceled) 
     
     
         8 . The system of  2  wherein the sample comprises tissue and wherein processing the sample comprises performing tissue dispersion. 
     
     
         9 . The system of  claim 1  wherein the sample comprises at least one of fresh tissue, cryogenically preserved tissue, and fixed and paraffin embedded tissue. 
     
     
         10 . The system of  claim 1  wherein the container includes a plurality of outlets for delivering the solution of microbubbles or the solution that will formulate encapsulated microbubbles when processed to different wells of a multi-well plate. 
     
     
         11 . The system of  claim 1  wherein a majority of the microbubbles have a diameter in the range from 0.1 microns to 10 microns. 
     
     
         12 . The system of  claim 1  comprising a second container for holding dehydrated microbubbles, which are mixed with a solution and thus re-suspended, and that provides the solution of microbubbles to the container for holding the solution of microbubbles. 
     
     
         13 . (canceled) 
     
     
         14 . The system of  claim 1  comprising a microbubble forming apparatus for processing the solution to form encapsulated microbubbles within the solution that will formulate encapsulated microbubbles when processed. 
     
     
         15 - 24 . (canceled) 
     
     
         25 . The system of  claim 1  where the solution that will formulate encapsulated microbubbles when processed includes surfactants, emulsifiers, polymers, or proteins. 
     
     
         26 . The system of  claim 14  wherein the microbubble forming apparatus comprises a probe for adding sonic energy to the solution that will formulate encapsulated microbubbles when processed in the presence of gas to induce the formation of the microbubbles in the solution before it is added to the biological sample. 
     
     
         27 . The system of  claim 14  wherein the microbubble forming apparatus comprises a mechanical mixer or impeller that rapidly agitates the solution that will formulate encapsulated microbubbles when processed in the presence of gas, forming the microbubbles in the solution, before the solution is added to the biological sample. 
     
     
         28 . The system of  claim 14  wherein the microbubble forming apparatus comprises at least one fluid system that forces gas surrounded by liquid through an opening to result in microbubble production. 
     
     
         29 . The system of  claim 14  wherein the microbubble forming apparatus comprises at least one fluid system that forces gas through a porous membrane to form the microbubbles. 
     
     
         30 . The system of  claim 1  wherein the solution that will formulate encapsulated microbubbles when processed comprises encapsulated liquid droplets, optionally wherein a majority of the droplets have a diameter in the range from 100 nanometers to 750 nanometers. 
     
     
         31 . The system of  claim 30  wherein the droplets comprise a shell surrounding a liquid core that converts to a gas upon the addition of acoustic, thermal, or optical energy. 
     
     
         32 . The system of  claim 31  wherein the liquid core comprises a hydrocarbon or perfluorocarbon, optionally at least one of isopentane, perfluoropentane, perfluorohexane, perfluorobutane, and perfluoropropane. 
     
     
         33 - 34 . (canceled) 
     
     
         35 . The system of  claim 30  comprising an energy source for converting the droplets into microbubbles, optionally wherein the energy source is configured to convert the droplets into microbubbles before or after the solution is delivered to the biological sample. 
     
     
         36 . (canceled)