The present invention relates generally to manufactured seeds and, more particularly, to a system and method for the delivery of plant embryos to a manufactured seed coat.
Modern agriculture, including silviculture, often requires the planting of large numbers of substantially identical plants genetically tailored to grow optimally in a particular locale or to possess certain other desirable traits. Production of new plants by sexual reproduction can be slow and is often subject to genetic recombinational events resulting in variable traits in its progeny. As a result, asexual propagation has been shown for some species to yield large numbers of genetically identical embryos, each having the capacity to develop into a normal plant. Such embryos must usually be further cultured under laboratory conditions until they reach an autotrophic “seedling” state characterized by an ability to produce their own food via photosynthesis, resist desiccation, produce roots able to penetrate soil and fend off soil microorganisms.
Some researchers have experimented with the production of artificial seeds, known as manufactured seeds, in which individual plant somatic or zygotic embryos are encapsulated in a seed coat, such as those disclosed in U.S. Pat. No. 5,701,699, issued to Carlson et al., the disclosure of which is hereby expressly incorporated by reference.
Typical manufactured seeds include a seed coat, a synthetic gametophyte and a plant embryo. Typically, the seed coat is a capsule having a closed end and an open end. The synthetic gametophyte is placed within the seed coat, such that the gametophyte substantially fills the seed coat. A cotyledon restraint may be centrally located within the synthetic gametophyte. The cotyledon restraint includes a centrally located cavity extending partially through the length of the cotyledon restraint and sized to receive the plant embryo therein. The well-known plant embryo is approximately 4-7 millimeters in length and roughly 0.5 millimeters in diameter. The shape of the plant embryo is somewhat cylindrical, but is irregular in cross-section and varies in diameter along its length. The plant embryo includes a radicle end and a cotyledon end. The plant embryo is deposited within the cavity of the cotyledon restraint cotyledon end first. The plant embryo is typically sealed within the seed coat by at least one end seal.
In the past, delivery of the plant embryo within the seed coat has utilized either conventional manually operated tweezers or vacuum pick-up devices to transfer the plant embryo through the manufactured seed production line. In such transfer systems that utilize conventional tweezers, the plant embryos are placed manually in separate seed coats, one at a time, by technicians. In such transfer systems that utilize vacuum pick-up devices, the plant embryos one at a time are grasped at their sides from a first position and transferred to a second position by an automated robotic arm. Attached to the end of the robotic arm is a pick-up head to which a source of vacuum to connected. The pick-up head includes a tip having a tip opening designed to grasp and hold a single plant embryo via vacuum pressure. After the pick-up head grasps the embryo, the embryo is positioned to acquire its morphological measurements and the location measurements for the radicle end. Then, the embryo is repositioned so that the embryo is held at the radicle end of the embryo, and is subsequently transferred to the second position for placing the embryo into the seed coat. Once the robotic arm is moved to the second position, the source of vacuum is shut off to release the embryo.
Although such plant embryo delivery systems are effective at transporting plant embryos, they are not without their problems. For example, when using conventional manually operated tweezers, the amount of force applied to the embryos is difficult to control. This results in the possibility of damaging the embryos, and the implementation of force sensors for such a small object using conventional methods to overcome this deficiency is too impractical for commercial success. When using vacuum pick-up heads, the embryo is not always successfully grasped due to the random orientation of the embryos and the variability of the size and shapes of the embryos. Additionally, the embryo surface is curved, which can prevent an adequate seal with the pick-up head tip opening. Such an imperfect seal may allow sufficient air flow around the embryo, resulting in a deficient vacuum to form. Accordingly, a lack of suction force is present to grasp and hold the embryo during the transfer process, which leads to unsuccessful transfers. Unsuccessful transfers of viable embryos are costly in modern automated material handling systems.
Secondly, with both aforementioned transfer methods, a problem may exist when either the operator or the automated pick-up head attempts to release the embryo into the seed coat. Specifically, since the embryos are kept moist or wet to prevent damage from desiccation, the embryo may remain attached to the tip of either the tweezers or the pick-up head due to the surface tension formed between the moisture on the embryo and the contact area of the tweezers or the pick-up head tip. In the case of conventional tweezers, to release the embryo, the technician typically positions the embryo to contact the side of the cotyledon restraint opening to create surface tension therebetween to overcome the surface tension associated with the tweezer tips. In the case of the vacuum pick-up head, a puff of air pressure is expelled out of the tip opening to overcome the surface tension and to force the embryo out of the vacuum head. In some instances, the burst of air flow is either insufficient to release the embryo or too great, in which case, the embryo is damaged by the impact force of the embryo against the bottom of the restraint. In either case, viable embryos may be wasted, which is costly in commercial applications. Further, the effects of surface tension and the conventional methods for overcoming the same may cause unwanted movement of the embryo, which in turn, affects the orientation of the embryo for insertion into the seed coat, and may lead to improper placement of or damage to the embryo.
The present invention is directed to an embryo delivery system that addresses the deficiencies of the prior art and others by employing automated microtweezers in embryo transfer process. The microtweezers, as will be described in detail below, are specifically designed to reduce the contact area of the tweezer tips on the embryos for reducing the surface tension therebetween. The reduction in surface tension results in improved embryo release capabilities for the embryo delivery system.
In accordance with one embodiment of the present invention, a method is provided for delivering embryos. The method includes positioning at least one embryo located on a support surface in a retrieval position. The oriented embryo is retrieved with automated microtweezers by actuation of the microtweezers to a closed position. The microtweezers are movable between a retrievable position and a release position. The automated microtweezers are moved to the release position where a seed coat is positioned relative to the release position. The embryo is then released into the seed coat by actuation of the microtweezers to an open position.
In accordance with another embodiment of the present invention, a method is provided for delivering plant embryos to a growing medium. The method includes imaging a plurality of plant embryos supported on a first surface for obtaining at least one selected plant embryo attribute, and orienting one plant embryo in a predetermined retrieval position based on the plant embryo attribute. The oriented embryo is transferred with microtweezers from the retrieval position to a release position, and then released from the microtweezers into the growing medium at the release position.
In yet another embodiment of the present invention, a method for delivering cultivated embryos is providing in a material handling system having an first positioning table, a transfer device having microtweezers, and a second positioning table. The method includes positioning a surface having a plurality of randomly oriented embryos onto the first positioning table, and obtaining at least one attribute of the randomly oriented embryos. One of the plurality of embryos is then orientated according to the obtained attribute by controlled actuation of the first positioning table so that the embryo achieves a selected, repeatable retrieval position. The embryo is transferred from the surface with the automated microtweezers to a selected, repeatable release position spaced from the surface, and placed into a seed coat positionally controlled by the second positioning table.
The foregoing aspects and many of the attendant advantages of this invention will become more readily appreciated by reference to the following detailed description, when taken in conjunction with the accompanying drawings, wherein:
The present invention will now be described with reference to the figures where like numerals represent like elements.
Referring now to
The orientation assembly 22 further includes an imaging system 50 or other suitable system for obtaining attributes of the plant embryos 46. The imaging system 50 may obtain any number of plant embryo attributes, such as size, shape, axial symmetry, cotyledon shape or development, surface texture, color, etc. In one embodiment, the imaging system 50 obtains either size or size and shape measurements, and based on these measurements, the embryos 46 will be classified as unqualified or qualified plant embryos. To be classified as a qualified embryo, the measurements of the embryo should indicate, within a sufficient tolerance, that the embryo will fit into the opening 126 of a cotyledon restraint 128 (See
The aforementioned attributes are obtained by the imaging system 50 by first acquiring and then digitally storing, if necessary, images of the plant embryos 46 by a well known digital imaging camera 54. The acquired and digitally stored images are then processed by a software program executed by the computer 56 of the control system 28 (See
In an alternative embodiment, the plant embryos 46 may be qualified or otherwise determined to be suitable for germination based on other criteria, for example, surface texture, color, IR absorption or reflection, Beta ray absorption, axial symmetry, and cotyledon development or any other attribute generally measurable by camera-like sensing devices. To this end, the acquired and digitally stored images of the digital imaging camera 54 may be sent to the computer 56 of the control system 28 (See
It will be appreciated that other classification methods and systems may be practiced with the present invention for selecting qualified embryos. For example, the embryos may be classified by the multi-stage screening process disclosed in copending U.S. patent application Ser. No. 10/611,756, entitled: Automated System And Method for Harvesting and Multi-Stage Screening of Plant Embryos, filed Jun. 30, 2003, the disclosure of which is hereby incorporated by reference. Additionally, the embryos may be classified as qualified using a spectroscopic analysis method, such as IR spectroscopy, NIR spectroscopy, or Raman spectroscopy, as disclosed in PCT Application Serial No. PCT/US99/12128, entitled: Method for Classification of Somatic Embryos, filed Jun. 1, 1999. These classification methods may be applied to any absorption, transmittance, or reflectance spectra of the embryos to classify the embryos according to their chemical composition. Other methods using Raman spectroscopy for classifying embryos that may be practiced with the present invention are disclosed in copending U.S. patent application Ser. No. 10/611,530, entitled: Method For Classifying Plant Embryos Using Raman Spectroscopy, filed Jun. 30, 2003, the disclosure of which is hereby incorporated by reference. Further, the apical dome located at the cotyledon end of a plant embryo may be three dimensionally imaged and analyzed for classifying embryos as qualified. Some methods of three-dimensionally imaging an apical dome of a plant embryo can be found in copending U.S. patent application Ser. No. 10/611,529, entitled: Method and System For Three-Dimensionally Imaging an Apical Dome of a Plant, filed Jun. 30, 2003, which is hereby incorporated by reference.
In operation, once a plurality of embryos 46 are randomly positioned on the support surface 44, the imaging camera 54 of the imaging system 50 acquires images of the embryos 46 and transmits the images to the computer 56 (See
In accordance with one aspect of the present invention, the queuing order in which the qualified embryos 48 are selected for retrieval may be specifically determined for improving the throughput of the embryo delivery process. The retrieval order of the qualified embryos 48 from the support surface 44 may be determined by any number of throughput enhancement routines. In the preferred embodiment, the throughput enhancement routine is executed by the computer 56 (See
Referring now to
As best shown in
In operation, once the positioning table 40 orients one qualified embryo 48 into the retrieval position, the transfer assembly retrieves the qualified embryo 48. To do so, the transfer device 60 is translated along the track 62 and the microtweezers 80 are rotated by the rotary shaft 70 to the retrieval position, shown in phantom in
After the qualified embryo 48 is retrieved from the support surface 44, the transfer device 60 is translated along the track 62 to a second, release position, while contemporaneously rotating the shaft 70 in the direction shown by the arrow 92 and opposite of the retrieval direction. Due to the small size of the microtweezers 80 and the qualified embryo 48 to be retrieved, the imaging camera 54 may be operated continuously to provide feedback control information for repositioning the positioning table 40 and/or controlling the actuation of the microtweezers 80 via the computer 86 (See
While the transfer device 60 is shown linearly translating along the track 62, it will be appreciated that other methods for transferring the qualified embryos from the retrieval position to the release position are possible. For example, the transfer device 60 may employ a robotic swing arm that rotates about the Z-axis for moving the microtweezers between such known positions. Additionally, it will be appreciated that the housing 66 may be a robotic housing capable of movement in the X, Y, and Z directions, as well as rotating about the Z axis. The robotic housing of such a transfer device may be used in conjunction with or in the absence of the positioning table 40 for positioning the microtweezers to retrieve the selected qualified embryos.
Returning to
Located on top of the positioning table 100 is a receptacle tray 110. The receptacle tray 110 includes a plurality of cavities 114 extending vertically therethrough, only one being shown in
In operation, having the positional information of the cotyledon restraint opening 126 of the manufactured seed coat 120 and the positional information of the cotyledon end 58 of the qualified embryo 48 held by the microtweezers 80 above the positioning table 100, the positioning table 100 precisely adjusts or indexes the location of the receptacle tray 134, such that it moves the opening 126 of the cotyledon restraint 128 to the precise location of the qualified embryo 48 held by the microtweezers 80. At this point, the microtweezers 80 are actuated from the closed position to the open position, and the qualified embryo 48 is released from the microtweezers 80 into the cotyledon restraint 128 of the manufactured seed coat 120.
As was described above in an alternative embodiment, the housing 66 of the transfer device may be a robotic housing capable of movement in the X, Y, and Z directions. The robotic housing of such a transfer device may be used in conjunction with or in the absence of the positioning table 100 for moving the microtweezers into a position to release the qualified embryo into the seed coat.
The operation of the embryo delivery system 20 will now be described by referring to
If the embryos 46 are qualified to be placed in a manufactured seed, the positional information of each qualified embryo 48 is determined and is used to assemble an embryo retrieval queue. In one embodiment of the present invention, the qualified embryos 48 are sorted and arranged in the queue by rotational coordinate information. Once the control system 28 generates a retrieval queue, whether using a throughput enhancement routine or not, the first qualified embryo 48 is oriented by the positioning table 40, through control signals sent by the control system 28, to the precise retrieval position.
Contemporaneously with or sequentially after orientating the qualified embryo 48 to the retrieval position, the control system 28 sends controls signals to the transfer device 60 such that the transfer device 60 translates to the retrieval position and the rotary shaft 70 rotates the microtweezers 80 in the direction opposite the arrow 92 to the embryo retrieval position. Once the microtweezers 80 are in the retrieval position, the microtweezers 80 are actuated to the closed position, thereby grasping the qualified embryo 48 between the microtweezer tips 88. In one embodiment, to improve the accuracy of the retrieval process and to control the force applied to the qualified embryo 48, the imaging system 50 may be continuously acquiring images of the position of the microtweezer tips 88 with respect to the qualified embryo 48, for providing feedback control information to the computer.
After the qualified embryo 48 is retrieved from the support surface 44, the transfer device 60 is translated in the opposite direction along the track 62 to the release position, while contemporaneously rotating the shaft 70 in the opposite direction shown by the arrow 92. In the release position, the microtweezers 80 hold the qualified embryo 48 within a sensor field of the position sensor 124 for obtaining positional information of the cotyledon end 58 of the qualified embryo 48. As best shown in
As noted above, simultaneous with or prior to the acquisition of the positional information for the qualified embryo, a second imaging camera associated with the positioning table 100 may locate the position of the opening 126 of the cotyledon restraint 128 in the manufactured seed 120 located on the positioning table 100. Alternatively, the receptacle tray 110 may be oriented on the positioning table so that the positional information of the restraint opening 126 of each seed coat 120 may be obtained and stored by the control system. As a result, having both the positional information of the cotyledon restraint opening 126 of the manufactured seed coat 120 and the positional information of the cotyledon end 58 of the qualified embryo 48, the positioning table 100 then locates itself through control signals sent by the computer 56, to accurately and precisely align the qualified embryo 48 with the opening 126 of the cotyledon restraint 128.
Once the qualified embryo 48 in aligned with the opening 126 of the cotyledon restraint 128, the microtweezers 80 are actuated by the control system 28 to the open position, thereby releasing the qualified embryo 48 into the manufactured seed coat 120. As was described above, the tips 88 of the microtweezers 80 are configured to reduce the contact area against the qualified embryo 48. As such, the weight of the qualified embryo may overcome the surface tension generated between the moist qualified embryo and the contact area of the microtweezer tips 88, thereby releasing the qualified embryo 48 from the microtweezers 80. If for some reason the qualified embryo 48 remains coupled to the microtweezer tips 88, the positioning table 100 may be slightly jogged to release the qualified embryo 48 from the microtweezers 80.
The embodiments of the present invention provide several advantages over currently available embryo delivery systems, some of which will now be explained. First, by employing microtweezers, and controlling its actuation distance, the force exerted on the qualified embryos can be precisely controlled, minimizing potential damage to the qualified embryos. Secondly, by employing the microtweezers, the contact area of the tips of the microtweezers against the embryo is purposefully and significantly reduced as compared to prior art methods, which in turn, minimizes the surface tension forces between the microtweezer tips and the qualified embryo.
While the orientation assembly 22 in the embodiments shown in
While the preferred embodiments of the invention has been illustrated and described, it will be appreciated that various changes can be made therein without departing from the spirit and scope of the invention, as claimed.
The present application claims the benefit of U.S. Provisional Application No. 60/525,449, filed Nov. 25, 2004, under 35 USC §119(e).
Number | Date | Country | |
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60525449 | Nov 2003 | US |