METHOD AND APPARATUS FOR TISSUE TRANSFER

• US 20100248370A1
• Filed: 03/16/2010
• Published: 09/30/2010
• Est. Priority Date: 03/26/2009
• Status: Active Grant

• Current Assignee: Corteva Agriscience LLC (Corteva, Inc.)
• Original Assignee: Dow Agrosciences LLC (Corteva, Inc.)
• Inventors: Moynahan, Tonya Strange
• Granted Patent: US 8,293,532 B2
• Time in Patent Office: Days
• US Class Current: 435/420
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 3/0275 Interchangeable or disposable dispensing tips
• CPC Class Codes: B25J 15/0616 with vacuum
• CPC Class Codes: B25J 19/04 Viewing devices
• CPC Class Codes: B25J 9/1697 Vision controlled systems
• CPC Class Codes: C12M 33/04 by injection or suction, e.g. using pipettes, syringes, needles pipettes in general B01L3/02
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: Y10S 901/09 Closed loop, sensor feedback controls arm movement
• CPC Class Codes: Y10S 901/40 Vacuum or mangetic
• CPC Class Codes: Y10S 901/47 Optical

Abstract

A handheld tool is disclosed which may be used to transfer a plurality of plant tissue explants from a first container to a second container. The handheld tool may include a disposable tip member which couples the plurality of plant tissue explants through use of negative pressure. An automated system which transfers a plurality of plant tissue explants from a first container to a second container is also disclosed. The automated system may include a first presentment system which moves the first container to a region, a second presentment system which moves the second container to the region, and a robot system that transfers the plurality of plant tissue explants from the first container to the second container.

Claims

1. A method of transferring plant tissue explants from a first container having a first nutrient medium substrate adapted to provide nutrients to the plant tissue explants to a second container having a second nutrient medium substrate adapted to provide nutrients to the plant tissue explants, the method comprising the steps of:

(a) coupling a first plant tissue explant to a handheld tool, the handheld tool coupling the first plant tissue explant through a negative pressure present in an interior of the handheld tool;

(b) generally separating the first plant tissue explant from the first nutrient medium substrate adapted to provide nutrients to the first plant tissue explant and from the first container;

(c) locating the first plant tissue explant relative to the second nutrient medium substrate adapted to provide nutrients to the first plant tissue explant of the second container; and

(d) uncoupling the first plant tissue explant from the handheld tool.

2. The method of claim 1, wherein the handheld tool includes a fluid conduit having a first end and a second end, the second end of the fluid conduit being in fluid communication with the first end of the fluid conduit and the fluid conduit having at least one vent passage positioned between the first end of the fluid conduit and the second end of the fluid conduit and being in fluid communication with the first end of the fluid conduit and the second end of the fluid conduit and to the surrounding environment, the vent passage being blocked from fluid communication with the surrounding environment during steps (a) through (c) with a stop device and the vent passage being in fluid communication with the surrounding environment during step (d).

3. The method of claim 2, wherein stop device is one of an operator'"'"'s finger and a device supported by the operator'"'"'s finger.

4. The method of claim 2, wherein the fluid conduit is an assembly and includes at least a first portion and a second portion, both the first portion and the second portion having a respective interior in fluid communication with the first end of the handheld tool and the second end of the handheld tool when the first portion and the second portion are assembled, the method further comprising the steps of:

(e) providing a plurality of first portions of the handheld tool;

(f) assembling a first one of the plurality of first portions of the handheld tool to the second portion of the handheld tool;

(g) performing steps (a) through (d) for all of the plant tissue explants in the first container selected for transfer to the second container;

(h) separating the first one of the plurality of first portions of the handheld tool from the second portion of the handheld tool;

(i) obtaining a third container including another plurality of plant tissue explants for transfer to a fourth container;

(j) assembling a second one of the plurality of first portions of the handheld tool to the second portion of the handheld tool; and

(k) performing steps (a) through (d) for all of the plant tissue explants in the third container selected for transfer to the fourth container.

5. The method of claim 4, wherein the plurality of first portions are a plurality of pipette tips.

6. The method of claim 5, wherein the plurality of pipette tips are stored generally vertical in a pipette tip box with a tip portion lower than a coupling portion, the step of assembling a first one of the plurality of first portions of the handheld tool to the second portion of the handheld tool includes the step inserting a portion of the second portion of the handheld tool into the coupling portion of a first pipette tip of the plurality of pipette tips while the first pipette tip is stored in the pipette tip box.

7. The method of claim 1, wherein steps (a) through (d) are performed in an aseptic environment.

8. The method of claim 7, wherein steps (a) through (d) are performed in a laminar flow hood.

9. The method of claim 1, wherein the step of uncoupling the first plant tissue explant from the handheld tool includes the step of reducing the negative pressure in the handheld tool by opening a vent passage of the handheld tool.

10. A system coupled to a negative pressure source for use transferring plant tissue explants from a first container having a first nutrient medium substrate adapted to provide nutrients to the tissue explants to a second container having a second nutrient medium substrate adapted to provide nutrients to the plant tissue explants, the system comprising:

a flexible fluid conduit having a first end adapted to be coupled to the negative pressure source and a second end which is in fluid communication with the first end through an interior of the flexible fluid conduit;

a handheld pen shaped instrument coupled to the second end of the flexible conduit, the handheld pen shaped instrument having an interior in fluid communication with the interior of the flexible fluid conduit; and

a tip member removably coupled to the handheld pen shaped instrument, the tip member having an interior in fluid communication with the interior of the handheld pen shaped instrument and a first end having an opening in fluid communication with the interior of the tip member, the opening of the first end of the tip member being smaller than the plant tissue explants to prevent the plant tissue explants from entering the interior of the tip member when a negative pressure is present in the interior of the flexible fluid conduit, the handheld pen-shaped instrument, and the tip member.

11. The system of claim 10, wherein the handheld pen shaped instrument includes at least one vent passage along an exterior of the handheld pen shaped instrument and in fluid communication with the surrounding environment and the interior the handheld pen shaped instrument, wherein when the vent passage is not in fluid communication with the surrounding environment the negative pressure in the tip member is increased compared to when the vent passage is in fluid communication with the surrounding environment.

12. The system of claim 10, wherein the tip member is a pipette tip.

13. The system of claim 10, further comprising a filter positioned between the first end of the flexible fluid conduit and the first end of the tip member.

14. The system of claim 13, wherein the flexible fluid conduit is divided into a first portion and a second portion, the filter being housed in a housing which is interposed between the first portion and the second portion.

15. A system coupled to a negative pressure source for use transferring plant tissue explants from a first container having a first nutrient medium substrate adapted to provide nutrients to the tissue explants to a second container having a second nutrient medium substrate adapted to provide nutrients to the plant tissue explants, the system comprising:

a flexible fluid conduit having a first end adapted to be coupled to the negative pressure source and a second end which is in fluid communication with the first end through an interior of the flexible fluid conduit; and

a tip member removably coupled to the flexible fluid conduit, the tip member having an interior in fluid communication with the interior of the flexible fluid conduit and a first end having an opening in fluid communication with the interior of the tip member, the opening of the first end of the tip member being smaller than the plant tissue explants to prevent the plant tissue explants from entering the interior of the tip member when a negative pressure is present in the interior of the flexible fluid conduit and the tip member, the tip member further includes at least one vent passage along an exterior of the tip member and in fluid communication with the surrounding environment and the interior the tip member, wherein when the vent passage is not in fluid communication with the surrounding environment the negative pressure in the tip member is increased compared to when the vent passage is in fluid communication with the surrounding environment.

16. The system of claim 15, wherein the tip member is a pipette tip.

17. The system of claim 15, further comprising a filter positioned between the first end of the flexible fluid conduit and the first end of the tip member.

18. The system of claim 17, wherein the flexible fluid conduit is divided into a first portion and a second portion, the filter being housed in a housing which is interposed between the first portion and the second portion.

19. A system coupled to a negative pressure source for use transferring a plurality of plant tissue explants from a first container having a first nutrient medium substrate adapted to provide nutrients to the tissue explants to a second container having a second nutrient medium substrate adapted to provide nutrients to the plant tissue explants, the system comprising:

a robot system supporting a fluid conduit having an interior which is in fluid communication with the negative pressure source;

at least one camera positioned to monitor a region including the first container and the second container; and

a controller operatively coupled to the at least one camera and the robot system, wherein the controller based on input from the at least one camera (a) moves the robot system to couple a first tissue explant of the plurality of plant tissue explants in the first container to the fluid conduit through negative pressure in the interior of the fluid conduit, (b) moves the robot system such that the first tissue explant is proximate the second nutrient medium of the second container; and

(c) uncouples the first tissue explant from the fluid conduit such that the first tissue explant is left in the second container.

20. The system of claim 19, wherein the controller repeats steps (a) through (c) for each of the plurality of plant tissue explants in the first container.

21. The system of claim 20, wherein each of the plurality of plant tissue explants are placed in the second container to form a predefined pattern.

22. The system of claim 19, further comprising an identification system operatively coupled to the controller, the identification system including a reader which determines an identification of the first container and a marker which provides identifying indicia on the second container.

23. The system of claim 19, further comprising a first presentment system which places the first container in the region and a second presentment system which places the second container in the region.

24. The system of claim 23, wherein the first presentment system removes the first container from a first queue of containers for transfer and places the first container in a queue of waste containers subsequent to a completion of the transfer of plant tissue explants from the first container to the second container.

25. The system of claim 24, wherein the first presentment system is operatively coupled to the controller, the controller providing instructions to the first presentment system regarding the movement of the first container.

26. The system of claim 25, wherein the first presentment system includes a turntable which transports the first container from the first queue of containers for transfer to the region and from the region to the queue of waste containers.

27. The system of claim 25, wherein the second presentment system removes the second container from a second queue of containers for transfer and places the second container in a third queue of containers subsequent to a completion of the transfer of plant tissue explants from the first container to the second container.

28. The system of claim 27, wherein the second presentment system is operatively coupled to the controller, the controller providing instructions to the second presentment system regarding the movement of the second container.

29. The system of claim 28, wherein the second presentment system includes a turntable which transports the second container from the second queue of containers for transfer to the region and from the region to the third queue of containers.

30. The system of claim 21, wherein the fluid conduit includes a removable tip member which interfaces with the plurality of tissue portions.

31. The system of claim 30, wherein the controller prior to steps (a) through (c) selects the removable tip member from a queue of removable tip members and subsequent to completion of steps (a) through (c) discards the removable tip member and selects a second removable tip member from the queue of removable tip members.

32. A method of transferring plant tissue explants, the method comprising the steps of:

(a) monitoring a region with at least one camera;

(b) providing a first container with a plurality of plant tissue explants supported on a first nutrient substrate for transfer in the monitored region;

(c) providing a second container in the monitored region for receiving the plurality of plant tissue explants and supporting the plurality of plant tissue explants on a second nutrient substrate; and

(d) automatically transferring the plant tissue explants from the first container to the second container through negative pressure applied by a robot system.

33. The method of claim 32, wherein the robot system includes a fluid conduit in fluid communication with a source of negative pressure and the step of automatically transferring the plant tissue explants from the first container to the second container through negative pressure applied by a robot system includes the steps of:

selecting a tip member from a queue of tip members;

coupling the tip member to the robot system such that a first end of the tip member is in fluid communication with the fluid conduit;

for each plant tissue explant in the first container;

locating the respective plant tissue explant in the first container;

positioning the first end of the tip member proximate the respective plant tissue explant;

coupling the respective plant tissue explant to the first end of the tip member due to the first end of the tip member being in fluid communication with the source of negative pressure through a fluid conduit of the tip member;

moving the tip member and the respective plant tissue explant to a predefined location relative to the second container;

uncoupling the respective plant tissue explant from the tip member by changing the pressure in the fluid conduit of the tip member; and

uncoupling the tip member from the robot system and discarding the tip member.

34. The method of claim 33, wherein the step of providing a first container with a plurality of plant tissue explants supported on a first nutrient substrate for transfer in the monitored region includes the steps of:

identifying the first container;

selecting the first container from a first queue of containers;

removing a lid of the first container; and

moving the first container to the region being monitored.

35. The method of claim 34, wherein the step of providing a second container in the monitored region for receiving the plurality of plant tissue explants and supporting the plurality of plant tissue explants on a second nutrient substrate includes the steps of:

selecting the second container from a second queue of containers;

identifying the second container;

removing a lid of the second container; and

moving the second container to the region being monitored.