System for high throughput sperm sorting

Patent metadata

Field	Value
Publication number	US-9757726-B2
Application number	US-201313830316-A
Country	US
Kind code	B2
Filing date	Mar 14, 2013
Priority date	Mar 14, 2013
Publication date	Sep 12, 2017
Grant date	Sep 12, 2017

Abstract

This disclosure relates to a system for sorting sperm cells in a microfluidic chip. In particular, various features are incorporated into the system for aligning and orienting sperm in flow channels, as well as, for determining sperm orientation and measuring relative DNA content for analysis and/or sorting.

First claim

We claim:

1. A sperm sorting system comprising:
a substrate;
at least one flow channel formed in the substrate, the flow channel having a sample inlet that receives sample, the flow channel further comprising: an inspection region, a first outlet, a second outlet, and a core stream forming geometry, the core stream forming geometry further comprising: a lateral fluid focusing region on the same plane as the sample inlet, a first vertical fluid focusing channel, and a second vertical fluid focusing channel, wherein one of the first fluid focusing channel or the second vertical fluid focusing channel is in fluid communication with the flow channel from a second plane above the inlet and the other is in fluid communication with the flow channel from a third plane below the inlet, and wherein the second vertical fluid focusing channel is downstream of the first vertical focusing channel in the flow channel;
at least one diverting mechanism in communication with the at least one flow channel to selectively divert sperm in the at least one flow channel away from the first outlet;
an electromagnetic radiation source for illuminating sperm at the inspection region of the at least one flow channel;
an analyzer in communication with the detector to determine sperm characteristics;
a controller in communication with the analyzer for selectively activating the diverting mechanism based on measured sperm characteristics; and
a collection vessel in communication with the second outlet.

2. The system of claim 1 , wherein the at least one flow channel comprises multiple flow channels formed on a microfluidic chip.

3. The system of claim 2 , wherein the multiple flow channels comprises between 4 and 512 flow channels.

4. The system of claim 2 , wherein either sperm characterized as viable X-chromosome bearing sperm or sperm characterized as viable Y-chromosome bearing sperm are deflected to the second outlet of each flow channel.

5. The system of claim 4 , wherein the collection vessel comprises a common collection vessel in fluid communication with the second outlet of the multiple flow channels.

6. The system of claim 1 , wherein each flow channel further comprises a third outlet.

7. The system of claim 6 , wherein the sperm cells characterized as viable X-chromosome bearing sperm are diverted to one of the second outlet or the third outlet and sperm characterized as viable Y-chromosome bearing sperm are diverted to the other of the second outlet and the third outlet.

8. The system of claim 6 , wherein each second outlet of the flow channels are connected to a first common collection vessel.

9. The system of claim 6 , wherein each third outlet of the flow channels are connected to a second common collection vessel.

10. The system of claim 1 , further comprising a passive collection vessel in communication with the first outlet.

11. The system of claim 1 , further comprising a sheath inlet that receives sheath fluid.

12. The system of claim 11 , further comprising a sheath fluid recycling system comprising:
a transport mechanism in fluid communication with the passive collection vessel;
a fluid path providing fluid communication between the passive collection vessel and the sheath inlet; and
a particle concentrating device or a fluid removing system in the fluid path providing fluid communication between the passive collection vessel and the sheath inlet.

13. The system of claim 1 , wherein the at least one flow channel comprises multiple flow channels formed on a microfluidic chip and wherein at least a portion of the diverting mechanism is embedded within the microfluidic chip.

14. The system of claim 1 , wherein the at least one flow channel comprises multiple flow channels formed on a microfluidic chip and wherein at least a portion of the diverting mechanism is positioned on the exterior of the microfluidic chip.

15. The system of claim 1 , wherein the diverting mechanism comprises a side passage in fluid communication with the flow channel and in fluid communication with a volume of fluid through a flexible interface.

16. The system of claim 15 , wherein the fluid comprises one selected from the group consisting of: a gel, a liquid, and a gas.

17. The system of claim 15 , further comprising an actuator contacting a portion of the flexible interface, wherein the actuator is in communication with the controller.

18. The system of claim 17 , wherein the actuator is movable between a resting position and two or more activation positions while maintaining contact with the flexible interface.

19. The system of claim 18 , further comprising a third outlet and wherein particles passively flow to the second outlet and wherein actuator movement between the resting position and the first active position diverts particles to the first outlet and wherein actuator movement between the resting position and the second active position diverts particles to a third outlet.

20. The system of claim 18 , wherein the actuator is attached to the flexible interface.

21. The system of claim 18 , wherein the actuator is preloaded onto the flexible interface.

22. The system of claim 15 , further comprising a bimorph piezoelectric element.

23. The system of claim 22 , wherein the bimorph piezoelectric element comprises the flexible interface.

24. The system of claim 22 , wherein the bimorph piezoelectric element contacts the flexible interface.

25. The system of claim 22 , wherein the bimorph piezoelectric element is configured for deflection in two directions to divert sperm in the flow channel two directions.

26. The system of claim 1 , wherein the diverting mechanism comprises a transducer coupled to the flow channel.

27. The system of claim 26 , wherein the transducer comprises an ultrasonic transducer for diverting particles in the flow channel.

28. The system of claim 27 , wherein the ultrasonic transducer comprise an array of ultrasonic transducers and further comprising a driving element which times the activation of each transducer in the array to achieve the desired deflection.

29. The system of claim 28 , comprising a second array of ultrasonic transducers, wherein each array of ultrasonic transducers is located on opposite sides of the flow channel.

30. The system of claim 28 , wherein the array of ultrasonic transducers is configured to produce multiple standing waves.

31. The system of claim 28 , wherein the array of ultrasonic transducers are configured to maintain the trajectory of a sperm cell in the flow path towards the first outlet, deflect the trajectory of a sperm cell in a flow path towards the second outlet, or deflect the trajectory of a sperm cell in a flow path towards a third outlet.

32. The system of claim 26 , wherein the transducer is at least partially embedded in the substrate adjacent to the flow channel.

33. The system of claim 26 , wherein the transducer is placed in contact with an exterior surface of the substrate.

34. The system of claim 1 , further comprising one or more sources of electromagnetic radiation for deflecting sperm in the flow channel.

35. The system of claim 1 , further comprising beam shaping optics for manipulating electromagnetic radiation produced from the electromagnetic radiation source to in

Assignees

• Inguran Llc

Inventors

• Sharpe Johnathan Charles
• Sedoglavich Nemanya
• Buchanan Kris
• Morad Blair
• Perrault Jr Donald Francis
• Koksal Erin

Classifications

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• B01L2400/0677

  phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers · CPC title

• B01L2400/0622

  distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves · CPC title

• B01L3/502738

  characterised by integrated valves (throttle valves in microfluidic sample containers B01L3/502746) · CPC title

• G01N2015/1006

  for cytology · CPC title