Systems and methods of separating and singulating embryos

Patent metadata

Field	Value
Publication number	US-9688953-B2
Application number	US-201514665344-A
Country	US
Kind code	B2
Filing date	Mar 23, 2015
Priority date	Apr 2, 2014
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

Separating and singulating embryos employs a spray module configured to spray a plurality of embryos which are loaded on a porous substrate so as to separate and singulate the plurality of embryos, and a drying module configured to dry the plurality of separated and singulated embryos retained on the porous substrate while the porous substrate is moved across the drying module. A robotic arm operates to transfer the porous substrate from module to module, and a control device controls the operation of the system of separating and singulating embryos.

First claim

What is claimed:

1. A system for separating and singulating embryos, comprising:
a dispensing module configured to dispense a blank porous substrate mounted in a frame;
a loading module for loading a plurality of embryos onto the porous substrate;
a spray module configured to spray a plurality of embryos loaded on the porous substrate so as to separate and singulate the plurality of embryos;
a drying module configured to dry the porous substrate upon which the plurality of separated and singulated embryos are retained;
a bioreactor loading module configured for loading the porous substrate upon which the plurality of separated and singulated embryos are retained into a bioreactor; and
a robotic arm operable to transfer the porous substrate from the spray module to the drying module;
wherein the dispensing module, the loading module, the spray module, the drying module, and the bioreactor loading module are arranged consecutively in a line from left to right or from right to left so that the porous substrate can be linearly transferred from one end to the other end of a processing line.

2. The system of claim 1 wherein the spray module comprises:
a spray hood capable of being raised or lowered by a lift mechanism and capable of engaging around a frame in which the porous substrate is mounted to create an isolated spray environment when lowered; and

a spray nozzle mounted to the approximate center of the internal top surface of the spray hood.

3. The system of claim 2 wherein the spray distance between the spray nozzle and the top surface of the porous substrate is adjustable by raising or lowering the spray nozzle.

4. The system of claim 2 wherein the spray angle of the spray nozzle is in the range of from about 26 degrees to about 32 degrees.

5. The system of claim 2 wherein the spray pressure at the spray nozzle is in the range of from about 20 psi to about 50 psi.

6. The system of claim 2 wherein the spray time of the spray module is in the range of from about 2 to about 30 seconds.

7. The system of claim 1 wherein the porous substrate comprises a plurality of pores, each of the plurality of pores having an opening width of from about 500 microns to about 2000 microns.

8. The system of claim 7 wherein the open area percentage of the porous substrate is equal to or greater than about 55%.

9. The system of claim 1 wherein the system comprises two processing lines which are arranged opposite to each other.

10. The system of claim 1 , further comprising a control device configured to control the operation of the system, the control device being coupled to the dispensing module, the spray module, the drying module, the bioreactor loading module, and the robotic arm.

11. The system of claim 1 , further comprising an overhead vision mechanism configured to count or at least estimate the number of separated and singulated embryos, the overhead vision mechanism being centered over the porous substrate after it has been dried at the drying module.

12. The system of claim 11 wherein the overhead vision mechanism comprises:
an image generator configured to capture the image of the separated and singulated embryos retained on the porous substrate; and
a light source configured to illuminate the separated and singulated embryos retained on the porous substrate, the light source being positioned such that a beam of light produced by the light source is approximately centered about the center of the porous substrate after it has been dried at the drying module.

13. A method of separating and singulating embryos, comprising:
dispensing a blank porous substrate mounted in a frame from a dispensing module;
loading a plurality of embryos onto the blank porous substrate while it is transferred to a predetermined loading position at a loading module;
spraying the plurality of embryos loaded on the porous substrate so as to separate and singulate the plurality of embryos at a spray module;
transferring the porous substrate from the spray module to a drying module; and
drying the porous substrate upon which the plurality of separated and singulated embryos are retained at the drying module; and
loading the porous substrate upon which the plurality of separated and singulated embryos are retained into a bioreactor at a bioreactor loading module;
wherein the dispensing module, the loading module, the spray module, the drying module, and the bioreactor loading module are arranged consecutively in a line from left to right or from right to left so that the porous substrate can be linearly transferred from one end to the other end of a processing line.

14. The method of claim 13 wherein the spray module comprises:
a spray hood configured to be raised or lowered by a lift mechanism and engage around a frame in which the porous substrate is mounted to create a closed spray environment when lowered; and
a spray nozzle mounted to the approximate center of the internal top surface of the spray hood.

15. The method of claim 14 wherein the spray distance between the spray nozzle and the top surface of the porous substrate is adjustable by raising or lowering the spray nozzle.

16. The method of claim 14 wherein the spray angle of the spray nozzle is in the range of from about 26 degrees to about 32 degrees.

17. The method of claim 14 wherein the spray pressure at the spray nozzle is in the range of from about 20 psi to about 50 psi.

18. The method of claim 14 wherein the spray time of the spray module is in the range of from about 2 to about 30 seconds.

19. The method of claim 14 wherein the porous substrate comprises a plurality of pores, each of the plurality of pores having an opening width of from about 500 microns to about 2000 microns.

20. The method of claim 19 wherein the open area percentage of the porous substrate is equal to or greater than about 55%.

21. The method of claim 13 wherein two processing lines are arranged opposite to each other.

22. The method of claim 13 , further comprising controlling the operation of the processing line by a control device.

23. The method of claim 22 , further comprising counting or at least estimating the number of separated and singulated embryos by an overhead vision mechanism, the overhead vision mechanism being centered over the porous substrate after it has been dried at the drying module.

24. The method of claim 23 wherein the overhead vision mechanism comprises:
an image generator configured to capture the image of the separated and singulated embryos retained on the porous substrate; and
a light source configured to illuminate the separated and singulated embryos retained on the porous substrate, the light source being positioned such that a beam of light produced by the light source is approximately centered about the center of the porous substrate after it has been dried at the drying module.

25. A system for separating and singulating embryos, comprising:
a spray module configured to spray a plurality of embryos loaded on a porous substrate so as to separate and singulate the plurality of embryos;
a drying module configured to dry the porous substrate upon which the plurality of separated and singulated embryos are retained;
an overhead vision mechanism configured to count or at least estimate the number of separated and singulated embryos, the overhead vision mechanism being centered over the porous substrate after it has been dried at the d

Assignees

• Weyerhaeuser Nr Co

Inventors

• Swanda Anthony P
• Givens Henry M

Classifications

• C12N5/04

  Plant cells or tissues {(culture media C12N5/0025)} · CPC title

• B07B4/08

  while the mixtures are supported by sieves, screens, or like mechanical elements · CPC title

• C12M47/04 Primary

  Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04) · CPC title