CN117546832A - Carrier bar and carrier bar clamp - Google Patents

Abstract

The invention relates to a carrier rod (100) for carrying active cells before vitrification and/or during low-temperature freezing and/or thawing, the carrier rod being of an elongated shape and having a rear part adapted to be held and a front part for carrying active cells, wherein the carrier rod comprises a liquid exchange port (103) and a cell exchange tank (101), wherein the cell exchange tank is located at the front part of the carrier rod, the bottom of the cell exchange tank is sealed, the top of the cell exchange tank is provided with an open notch, and the side wall is communicated with the liquid exchange port through a pore channel (104). The invention also relates to a carrier rod clamp (111) which is provided with a base (113) and an opening and closing part (112) which can pivot around the base, the carrier rod is communicated with an external automatic liquid exchanging device through a pipe joint on the opening and closing part, and a sealing ring which is suitable for pressing against a liquid exchanging port (103) of the carrier rod is arranged around the pipe joint (114) at one side facing the liquid exchanging port.

Description

Carrier bar and carrier bar clamp

Technical Field

The present invention relates to a carrier bar for carrying living cells prior to vitrification freezing and/or during low temperature freezing and/or thawing, said carrier bar being of elongate shape and having a rear portion adapted to be held and a front portion for carrying living cells. The invention also relates to a carrier rod clamp which is used for communicating the carrier rod with an external automatic liquid exchange device when clamping the carrier rod.

Background

The active cells need to be vitrified and frozen to maintain the activity during preservation and then re-warmed and thawed during use. Before vitrification freezing, the environment of the active cells needs to be changed, and the cell culture solution is replaced by vitrification freezing solution. After the thawing by re-warming, the vitrification frozen solution is replaced by the cell culture solution. The active cells are immersed in a plurality of specific liquids according to a certain sequence in the vitrification freezing and re-warming thawing process, so that the environment outside the cells is gradually changed step by step through the diffusion of the liquids, and the cells are adapted to the slowly-changing external liquid environment in the liquid exchange process, so that the activity is maintained.

The current main fluid exchange method is still manual operation of embryologists, taking the current rattan adding (Kitazato) method used in hospitals as an example, and the standard operation flow is to sequentially transfer cells from a cell culture fluid to a plurality of balance solutions for a certain time through the operation of sucking and pushing out the cells by a suction tube, and finally transfer the cells to a vitrification freezing solution. After the surrounding of the active cells has been replaced with a vitrification freezing solution, the active cells can be rapidly frozen in liquid nitrogen at a rate of, for example, -23000 ℃/min. When the active cells need to be used, the active cells are thawed by re-heating. Firstly sucking active cells by a suction tube, putting the active cells into a thawing solution at 37 ℃ for 1 minute, then taking out the active cells from the thawing solution, sequentially transferring the active cells into a dilution solution and a washing solution for a certain time, and finally taking out the active cells again to reach a condition for use.

For example, for oocytes, one standard workflow is: the oocytes were placed in a petri dish, immersed in 20. Mu.L of a basic solution (cell culture solution), then 20. Mu.L of the equilibration solution was added to the petri dish, after 3 minutes, 20. Mu.L of the equilibration solution was added, after 3 minutes, 240. Mu.L of the equilibration solution was added, after 9 minutes, the oocytes were placed in 300. Mu.L of the vitrification frozen solution by pipetting for 0.5 minutes, and then placed in another 300. Mu.L of the vitrification frozen solution by pipetting for 0.5 minutes. When the oocyte is thawed by re-warming, firstly, the oocyte is sucked by a suction pipe, the oocyte is placed into a thawing solution at 37 ℃ for 1 minute, then the oocyte is taken out of the thawing solution by the suction pipe, and then is placed into 300 mu L of diluted solution for 3 minutes, then is placed into a washing solution for 5 minutes, then is placed into another washing solution for 1 minute, and then the oocyte is taken out and placed into a vessel for further use.

For embryonic cells, the standard workflow is, for example: embryo cells were placed in a petri dish, immersed in 300. Mu.L of equilibration solution for 12-15 minutes, then aspirated with a pipette, placed in 300. Mu.L of vitrification frozen solution for 0.5 minutes, and then aspirated with a pipette and placed in another 300. Mu.L of vitrification frozen solution for 0.5 minutes. When the embryo cells are thawed by re-warming, firstly, the embryo cells are sucked by a suction pipe and placed into a thawing solution at 37 ℃, after 1 minute, the embryo cells are taken out of the thawing solution by the suction pipe and placed into 300 mu L of diluted solution for 3 minutes, then placed into 300 mu L of washing solution for 5 minutes, then placed into 300 mu L of another washing solution for 1 minute, and then the embryo cells are taken out and placed into a vessel for further use.

The prior art has the defects that the manual liquid change needs to be operated by an experienced embryologist, which requires training and operation experience for 2-3 years, has high labor cost, and is highly dependent on the hand feeling of the embryologist in the liquid change process due to the size of the active cells, so that the active cells are damaged once the liquid change is carried out by mistake.

Since the operation time of the vitrification freezing and the rewarming thawing process of the active cells is very short (for example, the time of placing the activated cells into the thawing solution after sucking the activated cells by the suction pipe is only about 1 minute as described above), the operation can be made faster and faster manually, and the operation habit of the operator such as doctor is satisfied by directly placing the cells into the thawing solution by using the carrier bar for taking out the activated cells from the liquid nitrogen for storing the activated cells at a low temperature, the process of manually placing the carrier bar for the activated cells into the liquid nitrogen for vitrification freezing and storing the activated cells at a low temperature on the carrier bar, and taking out the carrier bar into the thawing solution for thawing the activated cells is not urgent at present. However, for the liquid exchange process before and after the vitrification freezing and the rewarming thawing mentioned in the standard working flow, the manual operation is not the optimal choice because of the multiple steps, the multiple types and the different amounts of the used liquids and the different residence time required by each step.

In this regard, some automatic liquid changing devices and methods have also been proposed in the prior art. The Gavi system of Genea Biomedx in australia allows automated pipetting before vitrification freezing and after thawing at re-temperature. For this purpose, the cells are manually fixed in a cell tank of a specially-made liquid exchange container (consumable material), and then a three-dimensional moving platform is used for controlling a liquid transfer gun to add and remove a balancing solution, a vitrification freezing solution, a cell culture solution and the like into the liquid exchange container, and the liquid exchange is realized by a diffusion principle. The liquid-changing container can be heat-sealed with a heat-sealing film, so that the container is isolated from the outside during low-temperature preservation. The problem is that the liquid changing container used as a consumable material in the system is completely different from a pipette for transferring cells and a carrier rod for low-temperature freezing placed in a low-temperature tank which are conventionally used by operators such as doctors, and the operation habit of the doctors is seriously changed. Moreover, the system is too expensive to manufacture due to the complex motion of the three-dimensional moving platform controlling the pipette.

The Sarah system of fertillesafe in israel achieves automatic liquid change and vitrification freezing before vitrification freezing and after rewarming and thawing, but the vitrification freezing process does not completely achieve automation, excluding heat-seal parts. The low-temperature preservation and the rewarming and thawing still need to be manually operated. The automatic liquid change mode before vitrification and after rewarming and thawing is realized by distributing different solutions (balancing solution, vitrification and thawing solution, cell culture solution, thawing solution, liquid nitrogen and the like) on a wheel disc capable of horizontally rotating. The living cells are loaded into a specially made pipette (consumable) mounted on a z-axis movement module that moves in a z-axis direction perpendicular to the surface of the wheel. The wheel transfers the corresponding solution to the position right below the suction pipe through rotation, and the suction pipe moves up and down along the z-axis direction under the action of the z-axis moving module so as to be immersed in and separated from different solutions, and the preset residence time in the different solutions is realized. The disadvantage of this system is that the various solutions placed on the wheel for immersion of the pipette with the active cells, if the different active cells do not share the solutions, it takes a lot of time to change the solutions each time, which is not suitable for continuous pipelining, but if the different active cells share the solutions, there is a potential risk of cross-contamination. In addition, the system is complex and costly due to the motion of the two components involved in the system, the z-axis movement module and the wheel.

In the existing automatic liquid changing process before various vitrification freezing and after re-thawing, whether active cells are continuously transferred between various liquids by using a suction tube or placed in a cell tank different from a common suction tube and a carrier rod, the active cells need to be transferred to a special carrier rod during low-temperature freezing, and the operation of transferring the active cells for many times is inconvenient and time-consuming for doctors, operators and the like.

Accordingly, it is desirable to provide a carrier rod capable of automating the liquid changing steps before vitrification freezing and after rewarming and thawing in cooperation with an automatic liquid changing device, while changing the operation habits of operators such as doctors as little as possible. On this basis, it is also desirable to be able to realize a carrier bar-reagent, avoiding the sharing of carrier bars or various reagent solutions for active cells, and avoiding possible cross-contamination from the flow path. Obviously, it is also desirable that the cost of the device be controlled at a reasonable level.

Disclosure of Invention

In view of the above technical problem, the present invention provides a carrier bar for carrying living cells before vitrification freezing and/or during low-temperature freezing and/or thawing, said carrier bar being of elongate shape and having a rear portion adapted to be held and a front portion for carrying living cells. The carrier bar includes: the cell liquid exchange device comprises a liquid exchange port and a cell liquid exchange tank, wherein the cell liquid exchange tank is positioned at the front part of the carrier rod, the bottom of the cell liquid exchange tank is sealed, the top of the cell liquid exchange tank is provided with an open notch, and the side wall of the cell liquid exchange tank is communicated with the liquid exchange port through a pore canal.

The carrying rod of the invention is communicated with the outside through the liquid exchange port, such as a liquid exchange channel of an automatic liquid exchange device. The advantage of being communicated with the only liquid exchange channel is that continuous connection can be kept in the liquid exchange process, time consumption of repeated mechanical connection is reduced, and system reliability is improved. It is, however, obvious that the structural design of the carrier rod according to the invention is not affected, and that the fluid exchange port of the carrier rod is designed to be capable of alternating communication with a plurality of fluid exchange channels, for example by means of a common through-hole connection or luer connection with a plurality of fluid supply guns, each providing a different fluid. In any event, liquid enters the channel through the channel via the port and enters the cell fluid reservoir through the channel, and when liquid is to be pumped out, the liquid flows in the opposite direction and flows out of the carrier rod via the channel and the port. The carrying rod is provided with one or more liquid exchange ports for liquid inlet and one or more liquid outlet, and the liquid exchange ports are communicated with the cell liquid exchange tank through one or more pore channels. Preferably, the carrier rod is provided with a single liquid exchange port which is used for liquid inlet and liquid outlet, and the liquid exchange port is communicated with the cell liquid exchange groove through one or more pore channels. It will be appreciated that the arrangement of a plurality of channels leading to the fluid exchange channels, provided that the fluid exchange ports form a unique external connection, enables the channels forming a parallel arrangement to provide fluid from the unique fluid exchange ports to or withdraw fluid from the cell exchange channels more quickly and evenly. However, it will be appreciated that it is also possible to arrange a plurality of fluid change ports on the carrier bar, which provides the possibility of the carrier bar communicating with a plurality of fluid supply guns for alternating fluid changes, or for the fluid changes that must be manually effected exclusively with one fluid change port and that can be automated with the other fluid change port. In order to cooperate with an external automatic liquid exchange device for feeding and extracting liquid under pressure for a carrier rod, in the case of such a carrier rod on which a plurality of liquid exchange ports are arranged, it is necessary to be able to close the respective liquid exchange ports in time so that only one liquid exchange port at a time is able to communicate with the external automatic liquid exchange device. Whether a plurality of liquid exchange ports for liquid inlet and a liquid exchange port for liquid outlet, a liquid exchange port for liquid inlet and a plurality of liquid exchange ports for liquid outlet, a plurality of liquid exchange ports for liquid inlet and a plurality of liquid exchange ports for liquid outlet, and only one liquid exchange port for liquid inlet and only one liquid exchange port for liquid outlet or only one liquid exchange port for liquid outlet are arranged, the liquid exchange ports are arranged, and are used for liquid inlet and liquid outlet, the liquid exchange ports are all feasible embodiments of the application.

Advantageously, the bottom of the cell exchange well is provided with a cell fixing portion designed to accommodate the living cells and to restrict movement of the living cells, but not to prevent liquid from entering the surrounding area of the living cells. The cell fixing portion may be located at any position within the cell exchange liquid tank, but it is preferable that the cell exchange liquid tank has an elongated shape extending in the longitudinal direction of the carrier rod, and the cell fixing portion is located on a side of the cell exchange liquid tank remote from the aperture. Because the liquid enters the cell replacement tank through the pore canal, the arrangement of the cell fixing part away from the pore canal ensures that the active cells fixed in the cell replacement tank are less disturbed by the flow of the liquid, and the liquid can be more smoothly diffused to the periphery of the active cells to replace the original liquid. Alternatively, when the cell changing tank is small in size, the cell fixing portion may not be disposed, and the cell changing tank may be sized to directly restrict the movement of the living cells.

The cell fixing section may be only a region defined in advance in the cell changing bath without any special structural feature. But advantageously the cell fixing portion is a recess recessed into the bottom of the cell exchange well. After the active cells are placed in the pits, the active cells can be always kept in the pits in the liquid exchange process. Alternatively, a circumferentially closed mesh or grid structure protruding from the bottom of the cell exchange well may be used as the cell fixing portion. At this time, the liquid may flow into or out of the surrounding environment of the living cells through the gaps of the mesh-like or grid-like structure. It will be appreciated that the cell fixing portion may also be formed by a combination of a well recessed into the bottom of the cell exchange well and a circumferentially closed mesh or grid-like structure protruding at the circumferential edge of the well. At this time, the pit may have a shallow size.

In addition, the bottom of the cell changing bath is preferably transparent so that an operator such as a doctor can easily observe the state of the living cells carried therein under a microscope without taking out the living cells from the carrying bar. That is, the active cells can be not only subjected to liquid change and low temperature freezing after being placed on the carrier rod, but also can be directly observed under a microscope, and no tools are required to take out and transfer the active cells in these processes. This significantly simplifies the operation and provides convenience for operators such as doctors.

In order to enable smooth supply of liquid to the cell exchange liquid tank independently of the applied pressure at the liquid exchange port, the channel has a constriction on the side close to the cell exchange liquid tank, the constriction being dimensioned to block the entry of cells into the cell exchange liquid tank.

The carrier rod is provided with an information area outside the cell liquid exchange groove and the liquid exchange port, and the information area is used for providing visual or machine-readable information, such as bar codes, two-dimensional codes and the like carrying unique identification information of the carrier rod. An operator such as a doctor can uniquely associate and track the carrier bar by reading information in the information area.

The carrier bar may be flat or cylindrical in elongate shape to provide a feel that approximates that of existing pipettes or carrier bars. The carrier bar may be a bar-like structure adapted for finger pinching. Preferably, the carrier bar may also have concave contours adapted for finger pinching at the end remote from the cell exchange well. For example, a pair of concave contour portions disposed on both sides of the carrier bar are adapted to be pinched with two fingers by an operator such as a doctor. The concave contour portion is disposed as close to the rear portion of the carrier bar as possible so that an operator such as a doctor can grip the carrier bar even when the carrier bar is placed in the cryotank. In the case of a smaller carrier bar, the above-mentioned concave contour may not be arranged on the carrier bar.

Preferably, the carrier bar has a flat front face, and the open notch at the top of the cell exchange well is flush with or at least partially protrudes beyond the front face of the carrier bar. The flush design facilitates the fitting of a sleeve, which will be described in more detail below, over the carrier rod. The design of the front surface of the partially protruding carrier rod is beneficial to limiting the overflow of the liquid in the inclined state of the carrier rod.

At least a portion of the information area is also disposed on the front face of the carrier bar for viewing by an operator such as a doctor and for reading information in the information area using a machine if necessary. It is obvious that the information area may also be arranged in other areas than the front side of the carrier bar, e.g. the machine readable information carried in the information area need not be arranged on the front side of the carrier bar, but only in a position oriented for machine reading, e.g. on the side remote from the operator of a doctor or the like, since this side is often closer to the external automatic liquid changing device for supplying and retrieving liquid.

The liquid exchanging port is arranged on the front surface of the carrying rod or the back surface opposite to the front surface. In the case of a carrier rod having a sufficient thickness, the liquid exchange port can also be arranged on the side of the carrier rod. The arrangement on the back or side of the carrier rod is advantageous in that the connection of the fluid exchange port to the external automatic fluid exchange device does not obstruct the view of the cell fluid exchange tank by operators such as doctors. The arrangement on the front side of the carrier rod is however more preferred, since this avoids the risk of liquid leakage, provides a convenient handling of the liquid change port by means of a common through-hole connection or luer connection, and is more advantageous in terms of processing, since the cell change tank and the liquid change port can then be formed simply by punching two through-holes in one blank and covering these two through-holes by means of the other blank, preferably transparent at least in the region of the cell change tank, the portholes then being enclosed jointly with the surface of the latter blank by means of a groove in the back side of the former blank.

According to a preferred embodiment of the carrier rod according to the invention, the fluid exchange port is designed for a common through-hole connection or luer connection, which communicates with a corresponding common through-hole connection or luer connection of the external automatic fluid exchange device. However, it is also possible to communicate with an external automatic liquid exchange device through a special carrier bar clamp. For example, according to another aspect of the present invention, there is provided a carrier bar clamp for communicating a carrier bar with an external automatic liquid changing device when clamping the carrier bar, having a base having a receiving portion adapted to receive the carrier bar with a notch of a cell liquid changing tank facing upward, and an opening and closing portion arranged on the base in a pivotable manner about the base, wherein a pipe joint penetrating from a side of the opening and closing portion facing the liquid changing port to a side facing away from the base is fixed on the opening and closing portion, the pipe joint being adapted to communicate with the external automatic liquid changing device on a side of the opening and closing portion facing the liquid changing port being adapted to be aligned with the liquid changing port of the carrier bar received in the receiving portion when the opening and closing portion is closed onto the base, the opening and closing portion being arranged with a seal ring around the pipe joint on a side facing the liquid changing port, the pipe joint being adapted to press against the carrier bar when the opening and closing portion is closed onto the base, so as to realize a seal against the periphery of the liquid changing port when the opening and closing the liquid changing port.

Obviously, the design of the opening and closing part can be changed according to the difference of the positions of the liquid exchange ports on the carrying rod relative to the cell liquid exchange groove. Under the condition that the cell liquid exchange groove and the liquid exchange port are both designed on the front surface of the carrier rod, and the opening and closing part is closed on the base, a gap penetrating from one side of the opening and closing part, which faces the liquid exchange port, to one side opposite to the base can be formed in the opening and closing part according to the requirement, and the size and the position of the gap are designed so as not to cover the notch and the bottom of the cell liquid exchange groove on the carrier rod when the opening and closing part is closed on the base. If the liquid exchange port of the carrier rod is not arranged on the front surface of the carrier rod together with the cell liquid exchange tank but on the side surface or the back surface of the carrier rod, or if the pivot shaft of the opening and closing part is not arranged on the side close to the cell liquid exchange tank but on the side far away from the cell liquid exchange tank or on the side surface of the carrier rod, the opening and closing part does not need to be designed with the above-mentioned gap, because the opening and closing part does not block the notch and the bottom of the cell liquid exchange tank in the pivoting opening and closing process. The non-blocking of the notch allows the active cells to be easily placed in the cell-changing tank, and the non-blocking of the bottom (in the case of a bottom made of transparent material of the cell-changing tank) facilitates observation of the active cells in the cell-changing tank by a microscope without having to take them out.

The support rod clamp has the advantages that the pipe joint is continuously communicated with the liquid supply channel of the automatic liquid exchange device on one side of the pipe joint opposite to the base, and the pivoting of the opening and closing part and the pressing of the sealing ring jointly realize quick connection and reliable sealing. Compared with luer connection, the carrier rod can be deployed more quickly, and the working efficiency is improved.

Drawings

The present invention will be described in detail with reference to the accompanying drawings. In the drawings:

fig. 1 schematically shows a schematic diagram of an automatic liquid changing device according to the invention;

fig. 2 schematically shows a perspective view of a carrier bar according to the invention;

fig. 3 schematically shows a longitudinal section perspective of a carrier rod according to the invention;

FIG. 4 schematically illustrates an enlarged view of a portion of the carrier bar within the dashed box of FIG. 3 in accordance with the present invention;

fig. 5 schematically shows an assembly view of a carrier bar according to the invention arranged on a carrier bar clamp.

Detailed Description

Fig. 1 schematically shows a schematic diagram of an automatic liquid exchange device 110 according to the invention. In which liquid reservoirs 3 to 8 for storing liquid and a waste liquid reservoir 11 for storing waste liquid are shown in dashed boxes. The automatic liquid changing device 110 communicates with the carrier rod 100 (not shown in this figure) through the liquid changing channel 10. The dashed box indicates that the containers together form an integrally replaceable container assembly.

As shown in fig. 1, the positive pressure device 1 in the pumping device supplies positive pressure to the liquid storage containers 3 to 8, and the negative pressure device 13 supplies negative pressure to the waste liquid channel 14. When the multi-channel switching valve 9 communicates the liquid exchanging channel 10 with any one of the liquid supply channels 23 to 28, the liquid in the corresponding liquid storage containers 3 to 8 is supplied to the liquid exchanging channel 10 and thus to the carrier rod. When the multi-channel switching valve 9 communicates the liquid exchange channel 10 with the liquid waste channel 14, the liquid in the carrier rod is drawn back through the liquid exchange channel 10 and discharged out of the automatic liquid exchange device through the liquid waste channel 14.

Fig. 2 schematically shows a perspective view of a carrier bar according to the invention. The overall profile of the carrier bar 100 is elongate, with a rear portion for gripping and a front portion for carrying living cells. The carrier rod has a cell changing fluid bath 101 at the front and an information area at the rear for labeling visually or machine-readable information. In the middle, preferably front middle, of the carrier rod, in the region closer to the cell changing tank 101, a changing port 103 is arranged for communication with the changing channel 10 of the automatic changing device 110. The cell changing tank 101 has an open notch at the top, i.e., the side facing the front of the carrying rod 100, for facilitating the doctor or operator to take and place the active cells with a tool such as a pipette, and has a bottom and a periphery sealed and a channel 104 communicating with the changing port 103 at the side of the bottom, as shown in fig. 3. As shown in detail in fig. 4, the channel 104 has a constriction 105 on the side close to the cell-exchange liquid tank 101, the constriction being dimensioned to block the passage of cells into the cell-exchange liquid tank, so that the liquid flowing under the action of the pumping means can be smoothly fed into the cell-exchange liquid tank 101 or withdrawn from the cell-exchange liquid tank 101.

A cell fixing portion 102, preferably a well away from the well 104 as shown in fig. 3, is provided in the cell exchange well 101. Thus, after the cells are placed in the cell fixing section 102 in the cell changing tank 101, the cells do not move along with the supply and extraction of the liquid.

As shown in FIG. 2, the carrier rod also has an information area 106 for providing visual or machine-readable information in the area outside the cell change channel and the change port. The information area 106 carries a unique identification of the carrier bar, such as a digital code and a machine-readable bar code or two-dimensional code. The digital code is located on the front surface of the carrier bar 100, and the machine-readable bar code or two-dimensional code may be additionally located on both sides of the carrier bar in addition to the front surface, so that an operator such as a doctor may directly read the digital code from above, and the external automatic liquid changing device 110 may obtain a field of view of the bar code or two-dimensional code on the side surface of the carrier bar, no matter in the case that the operator such as the doctor holds the carrier bar with the left hand or the right hand, thereby implementing recognition and tracking of the carrier bar.

To close the notch of the cell changing bath 101 of the carrier bar, the doctor or the like inserts the carrier bar 100 placed in the low temperature bath into a bar sleeve (not shown) in the low temperature bath.

The carrier bar may be a bar-like structure adapted for finger pinching. The carrier bar also has a concave profile 107 near the rear for finger pinching. Only a pair of concave contours 107 on either side of the carrier bar are shown in fig. 2, which are adapted for a doctor or other operator to pinch the carrier bar into the cryotank with two fingers.

The manner of fluid communication between the carrier rod 100 and the automatic liquid changing device 110 is not shown in the drawings. A direct luer connection is a simple rotation, but it is also conceivable to use a combination of a common through-hole connection and a carrier bar clamp 111 as shown in fig. 5, the carrier bar 100 being accommodated in a receptacle 116 of a base 113 of the carrier bar clamp 111 with the notch of the cell exchange well 101 facing upwards. The opening and closing portion 112 is arranged on the base 113 in a pivotable manner to be opened and closed about the base 113. The pipe joint 114 is fixed to the opening and closing part 112, penetrates from the side of the opening and closing part 112 facing the liquid exchanging port 103 to the side facing away from the base 113, communicates with the external automatic liquid exchanging device 110 at the side of the opening and closing part 112 facing the base 113, is adapted to align with the liquid exchanging port 103 of the carrier rod 100 accommodated in the accommodating part 116 when the opening and closing part 112 is closed onto the base 113, and the opening and closing part 112 is provided with a sealing ring (not shown) around the pipe joint 114 at the side facing the liquid exchanging port 103, the sealing ring being adapted to press against the liquid exchanging port 103 of the carrier rod 100 when the opening and closing part 112 is closed onto the base 113, so as to realize sealing against the periphery when communicating with the liquid exchanging port 103. The size and position of the recess 115 provided in the opening and closing part 112, extending from the side of the opening and closing part 112 facing the liquid exchange port 103 to the side facing away from the base 113, is designed so as not to obstruct the notch and bottom of the cell exchange liquid tank 101 on the carrier rod when the opening and closing part 112 is closed onto the base 113.

Examples

The oocyte is placed in the cell fixing part 102 in the cell changing groove 101 of the carrier rod 100 by a pipette, and then the changing port 103 of the carrier rod 100 is communicated with the changing channel 10 of the automatic changing device 110. Then, the cell culture solution is supplied from the automatic solution changing device 110 to the cell solution changing tank 101 of the carrier rod 100 via the solution changing port 103 to submerge the oocyte. Then, the automatic liquid changing device 110 is operated to withdraw the cell culture solution from the carrier bar 100, and then a frozen equilibrium solution is supplied into the cell changing tank 101 of the carrier bar 100 through the liquid changing port 103 to submerge the oocyte, and is maintained for 3 minutes. Then, the automatic liquid changing device 110 is operated to withdraw the freezing balance solution from the carrier rod 100, and then vitrified freezing solution is provided to the cell liquid changing tank 101 of the carrier rod 100 through the liquid changing port 103 to submerge the oocytes, and after the operation is maintained for 0.5 minutes, the automatic liquid changing device 110 provides prompt information of the completion of liquid changing, so that operators such as doctors can disconnect the liquid communication between the carrier rod 100 and the automatic liquid changing device 110 to remove the carrier rod 100 from the automatic liquid changing device 110, sleeve the carrier rod 100 on the rod sleeve, and then put the carrier rod 100 together with the oocytes carried by the carrier rod into a low-temperature tank to be rapidly frozen by liquid nitrogen at the speed of-23000 ℃/min.

When the thawing is required, the doctor or other operator first takes out the carrier rod 100 carrying the oocyte from the low temperature tank, removes the rod sleeve, then adds the thawing liquid of 37 ℃ to the cell changing tank 101 by the pipette gun, and after 1 minute of soaking, then connects with the automatic changing device 110 by the changing port 103. The residual thawing solution in the cell-exchanging groove 101 of the carrier bar 100 was withdrawn by the automatic liquid-exchanging device 110, and then the diluted solution was supplied to the cell-exchanging groove of the carrier bar 100 for 3 minutes, and then the diluted solution in the cell-exchanging groove 101 of the carrier bar 100 was withdrawn and the washing solution was supplied to the cell-exchanging groove 101 of the carrier bar 100 for 3 minutes. Then, the operator such as a doctor can take out the oocyte from the cell fixing part 102 in the cell changing bath 101 of the carrier rod 100 with a pipette for further use. The operator such as a doctor may optionally determine whether the wash solution remaining in the cell changing bath 101 of the carrier rod 100 is first withdrawn by the automatic liquid changing device 110 before taking out the oocyte by using the pipette, according to the operation habit.

From the above, it is apparent that it is not necessary to remove the cells from the cell-changing tank 101 of the carrier bar 100 with a pipette for placement in the cryopanel during the whole process of pre-vitrification freezing, low-temperature freezing and re-thawing. The notch of the cell changing bath 101 of the carrier rod 100 may be simply closed by the nesting of the rod cover before being placed in the cryotank. When the cell-exchanging groove 101 of the carrier bar 100 is removed from the low-temperature groove, the groove opening of the cell-exchanging groove is exposed.

The foregoing has disclosed preferred embodiments of the present invention, however, the spirit and scope of the present invention is not limited to the specific disclosure. Those skilled in the art can devise many more embodiments and specific applications that do not depart from the spirit and scope of the invention. Therefore, it should be understood that the detailed description and specific examples, while indicating the scope of the invention, are not intended to limit the scope of the invention.

List of reference numerals

100. Loading rod

101. Cell changing liquid tank

102. Cell fixing part

103. Liquid exchanging port

104. Duct channel

105. Constriction part

106. Information area

107. Concave profile portion

108. Front face of carrier bar

110. Automatic liquid changing device

111. Carrier rod clamp

112. Opening and closing part

113. Base seat

114. Pipe joint

115. The absence of

116. Housing part

1. Positive pressure device

2. Switch valve

3. 4, 5, 6, 7, 8 liquid storage container

9. Multi-channel switching valve

10. Liquid exchange channel

11. Waste liquid container

12. Switch valve

13. Negative pressure device

14. Waste liquid channel

23. 24, 25, 26, 27, 28.

Claims (18)

1. A carrier rod for carrying active cells before and/or during low temperature freezing and/or thawing, said carrier rod being of elongated shape and having a rear portion adapted to be held and a front portion for carrying active cells, characterized in that said carrier rod comprises a liquid exchange port (103) and a cell exchange tank (101), wherein said cell exchange tank is located at the front portion of said carrier rod (100), wherein the bottom of said cell exchange tank (101) is sealed, the top has an open notch, the side walls are in communication with said liquid exchange port (103) through a duct (104).

2. Carrier bar according to claim 1, characterized in that the carrier bar has one or more liquid exchange ports (103) for liquid intake and one or more liquid discharge, the liquid exchange ports (103) being in communication with the cell exchange tank (101) via one or more of the portholes (104).

3. Carrier bar according to claim 2, characterized in that the carrier bar has only one fluid exchange port (103), the fluid exchange port (103) being used for both fluid intake and fluid discharge.

4. A carrier bar according to claim 3, characterized in that the bottom of the cell changing tank (101) is transparent.

5. Carrier bar according to claim 4, characterized in that the bottom of the cell changing tank (101) is arranged with a cell fixation (102) designed to accommodate the living cells and to restrict their movement, but not to hinder liquid from entering the surrounding area of the living cells.

6. The carrier bar according to claim 5, characterized in that the cell exchange fluid bath (101) has an elongated shape extending in the longitudinal direction of the carrier bar, the cell fixing portion (102) being located at a side of the cell exchange fluid bath (101) remote from the porthole (104).

7. The carrier bar according to claim 6, wherein the cell fixing portion (101) is a recess recessed into the bottom of the cell changing tank (101).

8. The carrier bar according to claim 6, characterized in that the cell fixation part (101) is a circumferentially closed mesh or grid structure protruding from the bottom of the cell exchange well (101).

9. The carrier bar according to claim 6, wherein the cell fixing portion (101) is a combination of a recess recessed into the bottom of the cell exchange well (101) and a circumferentially closed mesh or grid-like structure protruding at the circumferential edge of the recess.

10. Carrier bar according to any one of claims 1 to 9, characterized in that the channel (104) has a constriction (105) on a side close to the cell-exchange liquid bath (101), the constriction (105) being dimensioned to block cells from entering the cell-exchange liquid bath (101).

11. The carrier bar according to any one of claims 1 to 9, characterized in that the carrier bar (100) further has an information area (106) outside the cell exchange well (101) and the exchange well (103) for providing visually visible or machine readable information.

12. Carrier bar according to claim 11, characterized in that the carrier bar has a flat front face, the open notch of the top of the cell exchange well (101) being flush with or at least protruding partially out of the front face of the carrier bar.

13. Carrier bar according to claim 12, characterized in that at least a part of the information area (106) is also arranged on the front side of the carrier bar (100), the liquid exchange port (103) being arranged on the front side of the carrier bar (100) or on the back side opposite to the front side or on the side of the carrier bar (100).

14. Carrier rod according to any of claims 1 to 9, characterized in that the fluid exchange port (103) is designed to be adapted for a luer connection.

15. The carrying bar according to any one of claims 1 to 9, characterized in that the carrying bar (100) is a bar-like structure adapted for finger pinching.

16. The carrier bar according to any one of claims 15, characterized in that the carrier bar (100) further has a number of concave contours (107) adapted for finger pinching at an end remote from the cell exchange well (101).

17. A carrier bar clamp (111) for communicating the carrier bar (100) with an external automatic liquid changing device when clamping the carrier bar (100) according to any of claims 1 to 16, characterized in that the carrier bar clamp (111) is provided with:

a base (113) having a receiving portion adapted to receive the carrier bar (100) with a notch of the cell changing bath (101) upward; and

an opening/closing portion (112) which is arranged on the base (113) in a manner of being pivotable to open and close about the base (113),

wherein a pipe joint (114) penetrating from one side of the opening and closing part (112) facing the liquid exchanging port (103) to one side opposite to the base (113) is fixed on the opening and closing part (112), the pipe joint is suitable for being communicated with an external automatic liquid exchanging device at one side of the opening and closing part (112) opposite to the base (113), one side of the opening and closing part (112) facing the liquid exchanging port (103) is suitable for aligning the liquid exchanging port (103) of the carrier rod (100) accommodated in the accommodating part (116) when the opening and closing part (112) is closed on the base (113),

wherein the opening and closing part (112) is arranged around the pipe joint (114) on the side facing the liquid exchanging port (103), and the sealing ring is suitable for pressing against the liquid exchanging port (103) of the carrying rod (100) when the opening and closing part (112) is closed on the base (113) so as to realize sealing to the periphery when the liquid exchanging port (103) is communicated.

18. The carrier bar clamp (111) of claim 17, wherein the opening and closing part (112) has a void (115) penetrating from a side of the opening and closing part (112) facing the liquid exchange port (103) to a side facing away from the base (113), the void not blocking a notch and a bottom of a cell liquid exchange tank (101) on the carrier bar (100) when the opening and closing part (112) is closed onto the base (113).