The present invention relates to an automatic freezing processing apparatus, an automatic freezing processing method, and a freezing vessel.
Technology for freeze-preservation of a cell (freezing target), such as a fertilized embryo or an egg, has been developed as technology useful in an infertility treatment. In freeze-preservation of cells, a target cell such as an embryo or an egg is placed in a freezing vessel, and the cell is subjected to a freezing pretreatment. The cell placed in the freezing vessel is then quickly frozen by liquid nitrogen. In the freezing pretreatment, the cell is immersed in an equilibration solution (ES) and a vitrification solution (VS). This achieves substitution and vitrification of a cell sap inside the cell.
When manually performing the freezing pretreatment, an embryologist traps a target at a tip by drawing in and out with the mouth using a glass Pasteur pipette connected to a mouthpiece by a tube, thus transporting the target to the equilibration solution and to the vitrification solution and washing the target. For the freezing pretreatment, for example, U.S. Pat. No. 9,826,733 discloses technology of injecting an equilibration solution into a hollow in which an embryo is placed, collecting the equilibration solution, injecting the vitrification solution, and collecting the vitrification solution.
In the technology as disclosed in U.S. Pat. No. 9,826,733, a freezing target (such as an embryo) may be collected together in collection of the equilibration solution or the vitrification solution. In contrast, if the operator such as an embryologist fears the collection of the freezing target and a solution to be collected in the colleting operation is collected insufficiently, the solution to be collected may remain in the freezing target as a foreign matter. This requires the operator to operate carefully, leading to a heavy burden on the operator.
The present invention has been made in view of the above circumstances. An object of the present invention is, in freeze-preservation, to provide technology of reducing a burden on an operator in a pretreatment of a freezing target while avoiding a situation where the freezing target is accidentally collected and a situation where a solution to be collected remains in the freezing target.
An automatic freezing processing apparatus according to an aspect of the present disclosure includes a vessel that stores a freezing target, a working unit configured for injection and discharge of a liquid into and from the vessel and for movement of the vessel, and a refrigerant container that stores refrigerant for freezing the freezing target. The vessel has, at a bottom thereof, an opening smaller in size than the freezing target. The working unit discharges the liquid from the vessel through the opening after the injection of the liquid into the vessel, and moves the vessel to the refrigerant container.
An automatic freezing processing method according to an aspect of the present disclosure includes: injecting a liquid into a vessel having, at a bottom thereof, an opening smaller in size than a freezing target; discharging the liquid from the vessel through the opening after injecting the liquid; and moving the vessel to a refrigerant container after discharging the liquid.
A freezing vessel according to an aspect of the present disclosure is a freezing vessel for immersing a freezing target into a liquid for a freezing pretreatment. The freezing vessel includes a placement portion that allows the liquid to pass therethrough and the freezing target to be placed therein.
The foregoing and other objects, features, aspects and advantages of the present invention will become more apparent from the following detailed description of the present invention when taken in conjunction with the accompanying drawings.
Embodiments of the present disclosure will now be described in detail with reference to the drawings. The same or corresponding parts have the same reference characters allotted, and description thereof will not be repeated.
<Automatic Freezing Treatment Apparatus>
As shown in
One or more caps 300 are placed in cap holder 10. One or more tubes 200 are placed in tube holder 20. Tube 200 is an example of the freezing vessel.
ES reservoir 30 stores an ES. VS reservoir 40 stores a VS. Each of the ES and the VS is an example of the liquid used in the pretreatment of the freezing target. Liquid nitrogen tank 50 stores liquid nitrogen. The liquid nitrogen is an example of the freezing refrigerant. In liquid nitrogen tank 50, one or more freezing canes 60 are immersed in the refrigerant. Each freezing cane 60 houses one tube 200.
Electric pipetter 70 includes one or more nozzles 71 and sends air to each of one or more tubes 200 through a corresponding one of one or more nozzles 71. Air is sent with a pump 151, which will be described below with reference to
<Tube>
The size of the gap between the meshes of bottom 202 is smaller than the size of the freezing target. In one implementation, the freezing target is a fertilized egg. The fertilized egg has a size of approximately 140 μm, and each of the vertical length and the horizontal length of the gap is smaller than 140 μm (e.g., several tens of micrometers). Each of the vertical length and the horizontal length of the gap may be not greater than half the size of the fertilized egg (e.g., approximately 50 to 70 μm).
The gap between the meshes of bottom 202 is an example of the opening of tube 200. As long as bottom 202 of tube 200 has an opening, bottom 202 is not required to have a net structure.
Bottom 202 of tube 200 causes a liquid such as ES and VS to pass therethrough while holding freezing target 900. This eliminates the need for the operator to manually remove the liquid from tube 200 with freezing target 900 immersed in tube 200. Thus, such a situation can be avoided in which the liquid is removed insufficiently from tube 200 as the operator fears removal of freezing target 900.
<Cap>
Bottom 302 has a net structure. The mesh of bottom 302 may have a structure similar to the mesh of bottom 202 of tube 200. The gap between the meshes of bottom 302 is an example of the opening of cap 300. As long as cap 300 has an opening, cap 300 is not required to have a net structure at bottom 302.
The outside diameter of base 301 near bottom 302 is nearly the same as the inside diameter of base 201 of tube 200 near upper end 203. In other words, cap 300 covers tube 200 as its portion near bottom 302 is fitted into upper end 203 of tube 200.
A thread groove may be formed in the outer surface of base 301 near bottom 302 and in the inner surface of base 201 of tube 200 near upper end 203. Cap 300 may be screwed into tube 200 according to the threaded structure, thereby being fixed to tube 200.
<Control Block>
CPU 121 comprehensively controls freezing apparatus 100. CPU 121 deploys a program stored in ROM 122 to RAM 123 and executes the program. ROM 122 stores a program in which the procedure of freezing apparatus 100 is described. RAM 123 serves as a work area when CPU 121 executes the program, and temporarily stores the program or data in execution of the program, or the like.
Input unit 131 accepts an input including an instruction to freezing apparatus 100 from the user. Input unit 131 is, for example, a keyboard, a mouth, or a touch panel. Display 132, which is a display device that displays various screens, displays a state of process in freezing apparatus 100. Display 132 includes, for example, a display such as a liquid crystal display or an organic electro luminescence (EL) display.
Freezing apparatus 100 further includes a pump 151, an X-axis direction motor 161, a Y-axis direction motor 162, and an eject motor 171.
Pump 151 is driven for discharging and sucking air through a nozzle 71. X-axis direction motor 161 is driven for moving electric pipetter 70 in the X-axis direction. Y-axis direction motor 162 is driven for moving nozzle 71 of electric pipetter 70 in the Y-axis direction. Freezing apparatus 100 is equipped with a member that forms a mechanism for coupling electric pipetter 70 to X-axis direction motor 161 and Y-axis direction motor 162.
Eject motor 171 is driven for detaching, from nozzle 71, cap 300 attached to the tip of each of one or more nozzles 71. Electric pipetter 70 is equipped with a member that forms a mechanism for detaching cap 300 from nozzle 71.
In freezing apparatus 100, control device 110, pump 151, X-axis direction motor 161, Y-axis direction motor 162, and eject motor 171 are housed in, for example, case 101.
<Flow of Treatment>
Referring to
In step SA20, freezing apparatus 100 performs equilibration of freezing target 900.
Referring to
Freezing apparatus 100 moves electric pipetter 70 with cap 300 attached to nozzle 71, as shown in
Freezing apparatus 100 then moves nozzle 71 downward (
Freezing apparatus 100 then moves nozzle 71 upward (
Returning to
Freezing apparatus 100 causes nozzle 71 to suck air when bottom 202 of tube 200 is positioned to face the ES as shown in
Returning to
Freezing apparatus 100 discharges air into tube 200 through nozzle 71 with ES 31 injected into tube 200 as shown in
Returning to
Referring to
Freezing apparatus 100 discharges the ES from tube 200 (
Returning to
When bottom 202 of tube 200 is positioned to face the VS as shown in
Returning to
Freezing apparatus 100 discharges air into tube 200 from nozzle 71 with VS 41 injected into tube 200 as shown in
Returning to
Freezing of freezing target 900 in step SA40 will be descried with reference to
After the discharge of the VS from tube 200 (
Freezing apparatus 100 then drives eject motor 171 while raising nozzle 71 upward, thereby detaching cap 300 from nozzle 71 (
Then, a lid 61 is attached to freezing cane 60 (
As described above, freezing apparatus 100 freezes a freezing target.
In the process described with reference to
In the process described with reference to
<Automatic Freezing Treatment Apparatus>
Compared with freezing apparatus 100 shown in
<Flow of Treatment>
Referring to
Referring to
In step SB120, freezing apparatus 100A injects the ES into tube 200. More specifically, freezing apparatus 100A moves nozzle 71 with chip 401 attached thereto upward, moves electric pipetter 70 to ES reservoir 30, lowers nozzle 71, and drives pump 151. Thus, chip 401 sucks the ES. Freezing apparatus 100A then moves nozzle 71 upward, moves electric pipetter 70 to tube holder 20, and lowers nozzle 71.
Freezing apparatus 100 then drives pump 151. Thus, chip 401 discharges the ES into tube 200, and the ES is injected into tube 200.
Returning to
In step SB140, freezing apparatus 100A discharges the ES from tube 200. In one implementation, freezing apparatus 100 inserts chip 401 into the upper portion of tube 200, as shown in
Returning to
Returning to
Referring to
In step SB220, freezing apparatus 100A injects the VS into tube 200. More specifically, freezing apparatus 100A moves nozzle 71 with chip 402 attached thereto upward, moves electric pipetter 70 to VS reservoir 40, lowers nozzle 71, and drives pump 151. Thus, chip 402 sucks the VS. Freezing apparatus 100A then moves nozzle 71 upward, moves electric pipetter 70 to tube holder 20, and lowers nozzle 71. Freezing apparatus 100 then drives pump 151. Thus, chip 402 discharges the VS into tube 200.
In step SB230, freezing apparatus 100A stirs the liquid inside tube 200 as in step SB130.
In step SB240, freezing apparatus 100A discharges the VS from tube 200 as in step SB140.
In step SB250, freezing apparatus 100A detaches chip 402 from nozzle 71 as in step SB150. Freezing apparatus 100A then returns control to
Returning to
In step SB40, freezing apparatus 100A freezes freezing target 900 in tube 200. Freezing apparatus 100A then ends the process of
In one implementation, in step SB40, freezing apparatus 100A houses tube 200 and cap 300 attached to nozzle 71 in freezing cane 60, as described with reference to
In the process described with reference to
[Variation]
[Aspects]
It will be appreciated by a person skilled in the art that the exemplary embodiments described above provide specific examples of the following aspects.
(Clause 1) An automatic freezing processing apparatus according to an aspect may include a vessel that stores a freezing target, a working unit configured for injection and discharge of a liquid into and from the vessel and for movement of the vessel, and a refrigerant container that stores refrigerant for freezing the freezing target. The vessel may have, at a bottom thereof, an opening smaller in size than the freezing target. The working unit may discharge the liquid from the vessel through the opening after the injection of the liquid into the vessel, and move the vessel to the refrigerant container.
With the automatic freezing processing apparatus according to clause 1, in freeze-preservation, a burden on the operator in the pretreatment of the freezing target is reduced while avoiding a situation where the freezing target is accidentally collected and a situation where a liquid to be collected remains in the freezing target.
(Clause 2) In the automatic freezing processing apparatus according to clause 1, the injection of the liquid into the vessel may include injection of the liquid into the vessel from above the vessel, and the discharge of the liquid from the vessel may include sending air to the vessel from above.
With the automatic freezing processing apparatus according to clause 2, the liquid moves to the vessel under its own weight, thus allowing for more reliable injection of the liquid into the vessel.
(Clause 3) In the automatic freezing processing apparatus according to clause 2, the injection of the liquid into the vessel from above the vessel may include injection of the liquid using a chip.
With the automatic freezing processing apparatus according to clause 3, the liquid is injected into the vessel more reliably.
(Clause 4) In the automatic freezing processing apparatus according to clause 1, the injection of the liquid into the vessel may include sucking the liquid into the vessel from a tank that stores the liquid.
With the automatic freezing processing apparatus according to clause 4, the liquid is injected into the vessel reliably by a suction strength.
(Clause 5) In the automatic freezing processing apparatus according to any one of clauses 1 to 4, the working unit may perform an operation for stirring the liquid in the vessel between the injection of the liquid into the vessel and the discharge of the liquid from the vessel.
With the automatic freezing processing apparatus according to clause 5, the liquid acts on the freezing target in the vessel more reliably.
(Clause 6) The automatic freezing processing apparatus according to clause 5 may further include a pump that discharges air to the vessel for the operation.
With the automatic freezing processing apparatus according to clause 6, the liquid is stirred readily.
(Clause 7) In the automatic freezing processing apparatus according to any one of clauses 1 to 6, the working unit may attach a cap to the vessel.
With the automatic freezing processing apparatus according to clause 7, leakage of the freezing target in the vessel from the vessel is avoided more reliably.
(Clause 8) In the automatic freezing processing apparatus according to clause 7, the cap may have an opening smaller in size than the freezing target.
With the automatic freezing processing apparatus according to clause 8, the liquid can be injected into the vessel with the cap attached thereto while avoiding a situation where the freezing target in the vessel leaks from the vessel.
(Clause 9) In the automatic freezing processing apparatus according to any one of clauses 1 to 8, the injection of the liquid into the vessel may include injection of an equilibration solution into the vessel, and injection of a vitrification solution into the vessel after discharge of the equilibration solution from the vessel.
With the automatic freezing processing apparatus according to clause 9, a burden on the operator is reduced in both equilibration and vitrification of the freezing target.
(Clause 10) An automatic freezing processing method according to an aspect may include: injecting a liquid into a vessel having, at a bottom thereof, an opening smaller in size than a freezing target; discharging the liquid from the vessel through the opening after injecting the liquid; and moving the vessel to a refrigerant container after discharging the liquid.
With the automatic freezing processing method according to clause 10, in freeze-preservation, a burden on the operator in the pretreatment of the freezing target is reduced while avoiding a situation where the freezing target is accidentally collected and a situation where a liquid to be collected remains in the freezing target.
(Clause 11) A freezing vessel according to an aspect is a freezing vessel for immersing a freezing target into a liquid for a freezing pretreatment. The freezing vessel may include a placement portion that allows the liquid to pass therethrough and the freezing target to be placed therein.
With the freezing vessel according to clause 11, in freeze-preservation, a burden on the operator in the pretreatment of the freezing target is reduced while avoiding a situation where the freezing target is accidentally collected and a situation where a liquid to be collected remains in the freezing target.
Although the present invention has been described and illustrated in detail, it is clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation, the scope of the present invention being interpreted by the terms of the appended claims.
Number | Date | Country | Kind |
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2022-013066 | Jan 2022 | JP | national |