Methods and systems for using encapsulated microbubbles to process biological samples

Patent metadata

Field	Value
Publication number	US-11485994-B2
Application number	US-201815990171-A
Country	US
Kind code	B2
Filing date	May 25, 2018
Priority date	Oct 4, 2012
Publication date	Nov 1, 2022
Grant date	Nov 1, 2022

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.

First claim

What is claimed is:

1. A method for using encapsulated microbubbles to process a biological sample to shear and extract random, unbiased DNA fragments from the sample, the method comprising:
creating a mixture comprising encapsulated nanodroplets mixed with a biological sample comprising DNA, wherein creating the mixture comprises adding a solution of encapsulated nanodroplets to the sample and mixing the solution with the sample;
adding energy to the mixture to cause at least some of the nanodroplets to form encapsulated microbubbles, which oscillate or burst and thereby process the sample and shear the DNA, and
extracting the sheared, random, and unbiased DNA fragments from the sample.

2. The method of claim 1 wherein a majority of the encapsulated microbubbles have a diameter in the range from 0.1 microns to 10 microns.

3. The method of claim 1 wherein the biological sample comprises cells and wherein processing the sample comprises effecting cell lysis.

4. The method of claim 3 wherein the cells comprise cells from a tissue culture, bacteria cells, or yeast cells.

5. The method of claim 1 wherein the sample comprises tissue and wherein processing the sample comprises performing tissue dispersion.

6. The method of claim 1 wherein the sample comprises at least one of fresh tissue, cryogenically preserved tissue, and fixed and paraffin embedded tissue.

7. The method of claim 6 wherein the sample comprises fixed and paraffin embedded tissue.

8. The method of claim 1 wherein adding the energy to the mixture comprises sonicating the mixture.

9. The method of claim 8 wherein sonicating the mixture includes applying energy having a frequency in the range from 0.01 MHz to 10.0 MHz.

10. The method of claim 1 wherein adding the energy to the mixture comprises exposing the mixture to laser light.

11. The method of claim 1 wherein the nanodroplets comprise a shell surrounding a liquid core which converts to a gas upon the addition of the energy, wherein the energy comprises at least one of acoustic, thermal, and optical energy.

12. The method of claim 11 wherein the liquid core comprises a hydrocarbon or a perfluorocarbon.

13. The method of claim 12 wherein the liquid core comprises at least one of isopentane, perfluoropentane, perfluorohexane, perfluorobutane, and perfluoropropane.

14. The method of claim 1 wherein a majority of nanodroplets have a diameter in the range from 100 to 750 nanometers.

15. The method of claim 1 wherein adding the energy comprises adding formation energy to a biological sample to induce the formation of encapsulated microbubbles in the biological sample and adding activation energy to cause at least some of the microbubbles to oscillate or burst.

16. The method of claim 15 wherein creating the mixture comprises adding a surfactant, emulsifier, polymer, or protein to the sample prior to or during the addition of the formation energy to enhance bubble stability so that the microbubbles persist until the addition of activation energy.

17. The method of claim 15 wherein creating the mixture comprises adding the nanodroplets to the sample prior to or during the addition of the formation energy, wherein, during administration of formation energy, at least some of the nanodroplets vaporize into the bubbles that will oscillate or burst in response to the addition of activation energy.

18. The method of claim 15 wherein adding the formation energy comprises applying laser light to the sample to form the microbubbles.

19. The method of claim 15 wherein adding the formation energy includes sonicating the sample to form the microbubbles.

20. The method of claim 19 wherein sonicating the sample includes applying sonic energy having a frequency in the range from 0.01 MHz to 10.0 MHz.

21. The method of claim 1 wherein adding the solution of encapsulated nanodroplets to the sample comprises adding the solution to a plurality of biological samples located in individual wells of a multi-well sample plate, wherein mixing the solution includes mixing the solution with the samples in the wells wherein adding the energy to the mixture includes adding the energy to each of the mixtures in the wells to perform high throughput processing of a plurality of biological samples.

Assignees

• Univ North Carolina Chapel Hill

Inventors

• Janzen William Perry
• Pattenden Samantha Gail
• Jayakody Chatura Nadisha
• Streeter Jason Eric
• Dayton Paul Alexander
• Wood Cameron Champion
• Shenoy Siddharth Kaup

Classifications

• C12N15/10

  Processes for the isolation, preparation or purification of DNA or RNA (chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34) · CPC title

• C12N15/1003

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• G01N2001/387

  mixing by blowing a gas, bubbling · CPC title

• G01N1/38 Primary

  Diluting, dispersing or mixing samples · CPC title

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title