Microfluidic system and method with focused energy apparatus

Patent metadata

Field	Value
Publication number	US-11796449-B2
Application number	US-201916279430-A
Country	US
Kind code	B2
Filing date	Feb 19, 2019
Priority date	Oct 30, 2013
Publication date	Oct 24, 2023
Grant date	Oct 24, 2023

Abstract

A method of inseminating an animal including flowing a stream of a population of sperm cells through a channel, differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells, selecting a desired subpopulation, ablating an undesired subpopulation, and collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together, wherein the collected population of sperm cells is used to fertilize an egg.

First claim

What is claimed is:

1. A method of producing a sperm cell composition, comprising the steps of
flowing a stream of a population of sperm cells through a channel;
differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells;
selecting a desired subpopulation;
ablating an undesired subpopulation at a predetermined delay time from selecting the desired subpopulation, wherein the predetermined delay time is 1 μs or longer; and
collecting both the subpopulations of sperm cells including the desired subpopulation and the ablated undesired subpopulation together.

2. A method of fertilizing one or more eggs comprising the steps of providing at least one egg obtained from a female mammal, providing the sperm cell composition of claim 1 from a male mammal of the same species as the female mammal, and mixing the one or more eggs with the sperm cell composition.

3. The method of claim 2 , wherein the female mammal is a cow or a heifer.

4. The method of claim 2 , wherein the male mammal is a bull or boar.

5. The method of claim 2 , further comprising implanting the at least one fertilized egg within the female animal within forty-eight hours following fertilization.

6. The method of claim 2 , further comprising obtaining genetic information of the sperm cells or the fertilized egg prior to implantation.

7. The method of claim 6 , wherein the implanting the fertilized egg within the female animal occurs within forty-eight hours following obtaining the genetic information of the fertilized egg.

8. The method of claim 6 , wherein obtaining the genetic information of the fertilized egg comprises removing at least one cell of the fertilized egg for analysis.

9. The method of claim 6 , wherein obtaining the genetic information of the fertilized egg comprises sequencing a portion of a DNA molecule or an RNA molecule of the fertilized egg.

10. The method of claim 6 , wherein obtaining the genetic information of the fertilized egg occurs within forty-eight hours following removing at least one cell of the fertilized egg for analysis.

11. The method of claim 10 , wherein obtaining the genetic information of the fertilized egg occurs within twenty-four hours following removing at least one cell of the fertilized egg for analysis.

12. A method of producing an embryo comprising using the sperm cell composition of claim 1 and an assisted reproductive technique.

13. The method of claim 12 , wherein the assisted reproductive technique is selected from the group consisting of in vitro fertilization (IVF), artificial insemination (AI), intracytoplasmic sperm injection (ICSI), multiple ovulation and embryo transfer (MOET), and other embryo transfer techniques.

14. The method of claim 13 , wherein a heifer or cow is inseminated using artificial insemination.

15. The method of claim 14 , wherein success rate for insemination is better than insemination comprising a physical separation of selected sperm cells from unselected sperm cells.

16. The method of claim 15 , wherein success rate for insemination of a heifer is at least about 60% on average.

17. The method of claim 15 , wherein success rate for insemination of the cow is at least about 37% on average.

18. The method of claim 1 wherein flowing the stream of the population of sperm cells through the channel comprises
orienting the sperm cells in the population in a particular position with respect to the channel;
aligning the sperm cells wherein the sperm cells are flowing singularly through the channel; and
exiting all the sperm cells through an output portion of the channel.

19. The method of claim 18 , wherein orienting the sperm cells and aligning the sperm cells comprise
compressing the stream by sheath fluid flowing along at least one side of the stream wherein the compressing further comprises intersecting at a substantially 90 degree vertical plane by the sheath fluid from above the stream.

20. The method of claim 19 , wherein the sheath fluid is in laminar flow with the stream of the population of sperm cells during the differentiating and the ablating steps.

21. The method of claim 18 , wherein orienting the sperm cells and aligning the sperm cells comprises compressing the sperm cells by an inertial force directed by a designed structure of the channel.

22. The method of claim 1 , wherein differentiating the sperm cells into two subpopulations of X-chromosome containing sperm cells and Y-chromosome containing sperm cells is accomplished via the steps of:
illuminating the sperm cells with a beam of light; and
detecting a difference between the desired subpopulation and the ablated undesired subpopulation.

23. The method of claim 22 , wherein the beam of light is from an LED light source.

24. The method of claim 22 , wherein the beam of light is from a detection laser.

25. The method of claim 22 , wherein the beam of light is split into multiple beams of light; and wherein each beam of light of the multiple beams of light is configured to illuminate a different sperm cell.

26. The method of claim 22 , wherein the beam of light is not split into multiple beams and the beam illuminates one sperm cell at a time.

27. The method of claim 22 , wherein the beam of light is emitted as a continuous wave.

28. The method of claim 22 , wherein the beam of light is emitted in pulses.

29. The method of claim 22 , wherein the difference comprises how the sperm cells reflect, refract, diffract, absorb, and/or emit the light.

30. The method of claim 22 , wherein a molecule of the sperm cell or a molecule associated with the sperm cell is excited by the beam of light and the molecule emits a second beam of light in a Stoke shift.

31. The method of claim 22 , wherein the difference reflects a difference in DNA concentration per cell.

32. The method of claim 31 , wherein the difference between the subpopulations is at least about 3.8% difference in DNA content.

33. The method of claim 1 , wherein selecting the desired subpopulation comprises gating each subpopulation based on a physical difference for each subpopulation; and
identifying the desired subpopulation.

34. The method of claim 33 , wherein gating each subpopulation and identifying the desired subpopulation comprises
correlating a graphic representation for each subpopulation; and
choosing or marking the desired subpopulation.

35. The method of claim 1 , wherein ablating the undesired subpopulation further comprises
damaging or killing the undesired subpopulation of sperm cells with a second beam of light in an area of the channel,
wherein a damaged or killed sperm cell is rendered unable to fertilize an egg.

36. The method of claim 35 , wherein the second beam of light is from a laser.

37. The method of claim 36 , wherein the laser is configured to provide a pulse of beam of light and wherein the laser is configured to recharge to damage or kill the next sperm cell that enters the area of the channel.

38. The method of claim 35 , wherein the beam of light is tightly focused with a blade-shape configured to cut across at least approximately 2 microns in length of the channel.

Assignees

• Abs Global Inc

Inventors

• Appleyard David
• Betthauser Jeff
• Faust Marjorie
• Larsen John
• Shao Guocheng
• Xia Zheng
• Zhou Yu

Classifications

• G01N15/1484 Primary

  microstructural devices · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title

• C12N13/00

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• G01N1/44

  Sample treatment involving radiation, e.g. heat · CPC title

• G01N15/1404

  Handling flow, e.g. hydrodynamic focusing · CPC title