CN103444699B

CN103444699B - A kind of miniature base system, well-bucket and the cryopreservation methods frozen for biomaterial

Info

Publication number: CN103444699B
Application number: CN201310333111.XA
Authority: CN
Inventors: 吕祁峰
Original assignee: Individual
Current assignee: Individual
Priority date: 2013-08-01
Filing date: 2013-08-01
Publication date: 2015-07-29
Anticipated expiration: 2033-08-01
Also published as: CN103444699A

Abstract

The invention provides a kind of miniature base system frozen for biomaterial and application thereof, form primarily of frozen unit, described frozen unit comprises a year bar, carries bar cover and carry bar cylinder; Carry in bar front end insertion year bar cover and form smooth elongated load units; be loaded into together again and carry in bar cylinder; described year bar cylinder is the blank pipe of at least bottom end closure, mean outside diameter < 8.5mm, and the top or the sidewall that carry bar cylinder are provided with the dress bar mouth and loading protection thereof and taking-up technology that load units can be put into and taken out from year bar cylinder.Miniature base system provided by the invention makes, not affecting the space basis even more increasing ease of use decreased exponentially shared by micro-Frozen Biological, significantly to have saved the floor space needed for stored frozen.Be particularly useful for frozen as embryo of micro-biomaterial.

Description

Micro carrier system for cryopreservation of biological materials, bucket and cryopreservation method

Technical Field

The invention relates to a micro carrier system for freezing and storing biological materials, in particular to a set of sample loading device used for carrying out micro biological material vitrification freezing or other freezing in the biological and medical fields and application thereof.

Background

There are many known frozen carrier systems, which can be classified into a freezing tube type and a freezing rod type according to the main loading unit. The former is a generally cylindrical or partially cylindrical tubular container with a blind end at one end and a cap to close the other end in a threaded or snap-fit manner. Such containers are generally large, typically over 1 ml in volume, and are stored in tiered racks in liquid nitrogen tanks, mostly by box-pack loading. Most of the current cell cryopreservation takes this form; the latter is then a rod-like device comprising: (1) the device comprises (1) a long and thin straw-shaped container which can be used for freezing and storing 1-5 hundred microliter of biological material samples, (2) a long and thin pipe which is further stretched at the tail end and can be used for freezing and storing a few microliter of samples by using a long and thin part, and (3) a long and thin rod-shaped non-container which is provided with a freezing and storing slide at the front end and can be used for adhering and storing biological materials within a few microliter. The latter two forms belong to micro-sample cryopreservation, and are more applied to vitrification cryopreservation of ova or embryos cultured in vitro. For the vitrification freezing of the ovum or embryo, there is also micro freezing in the form of freezing ring, that is, the liquid adhered to the microfilament ring is used to load the ovum or embryo, which is then put into a tubular container for protection and freezing.

However, the conventional devices for freezing and storing various micro (within several microliters) samples do not occupy relatively less storage space than the conventional devices (several milliliters of micro-liters) which causes the application efficiency of the storage space to be low. The reasons for this are: the loading and taking of a trace sample also needs to meet the requirements of simple identification, convenient operation and the like, so that besides the loading part of the sample carrier, other related structures and devices are also needed to help the loading, sealing, marking, classified storage, convenient access and the like of the sample, and the combination of the structures and the devices forms a larger space at present.

Taking the example of freezing the embryo cultured in vitro in the human assisted reproduction technology, the vitrification freezing of the embryo usually adopts the form of a freezing rod (hereinafter, referred to as embryo freezing rod or rod carrier for short), such as a slender rod-shaped non-container form, which is the most space-saving and convenient form for embryo freezing at present, and includes a rod carrier (with a freezing piece at one end), a rod carrier sleeve, a rod carrier cap and a rod carrier system. Fig. 1 is a schematic diagram of a conventional carrier system, which is composed of a carrier rod 1 ', a carrier rod sleeve 2 ', a carrier rod barrel 3 ', a carrier rod barrel cap 4 ' and a carrier rod barrel frame 5 '. As shown in figure 1, the front end of the carrier rod 1 'is inserted into the carrier rod sleeve 2' and then is loaded into the carrier rod barrel 3 ', and the carrier rod barrel 3' is clamped on the carrier rod barrel frame 5 'and is covered by the carrier rod barrel cap 4' clamped by the carrier rod barrel frame 5 'to prevent the carrier rod 1' from falling out. The freezing storage piece on the carrying rod is used for placing embryos, and one end of the carrying rod, where the embryos are placed, is sleeved with the carrying rod sleeve so as to protect the section from being damaged by external force; the rod carrying cylinder is used for carrying the carrying rod and the rod carrying sleeve thereof; and an aluminum alloy frame, namely a rod carrying cylinder frame, which is used for clamping and fixing the rod carrying cylinder and a rod carrying cylinder cap (the rod carrying cylinder cap is used for protecting articles in the cylinder from falling out of the cylinder). The carrying rod can be pasted with a biological material information label. The upper end of the rod-carrying barrel frame can be pasted with information labels such as a freezing position and the like to form a freezing unit. Multiple sample carrying rods from the same patient are typically loaded into the same or multiple cryopreservation units, and embryos from different patient sources cannot be mixed in the same cryopreservation unit, so that identity marking and identification are facilitated.

The carrier rod holder and its clamped carrier rod and carrier rod cap as described above not only have a thickness of about 1-1.5 cm in diameter, but also have a total length of about 29 cm. In this set of devices, there are two aspects of creating maximum space usage: (1) the rod-carrying cylinder frame is characterized in that the rod-carrying cylinder frame is a rod-carrying cylinder frame, the rod-carrying cylinder frame is larger than a rod-carrying cylinder and can be wrapped around a rod-carrying cylinder cap which is used for clamping the rod-carrying cylinder and is as large as the rod-carrying cylinder, and the clamping is necessary in that the rod-carrying cylinder cap is fixed to be positioned at the opening end of the rod-carrying cylinder so as to prevent a rod-carrying in the rod-carrying cylinder from falling out; (2) is a rod carrier sleeve because it must be larger than the end of the rod on which the embryo is placed to fit over the end, often creating a larger footprint than the rod. This makes the rod carrier necessary to have a relatively large (about 1 cm) internal diameter to meet the usual loading (3-4 rods for 6-8 embryos).

Generally, at least one rod carrying barrel frame is needed for embryos of one patient, the freezing storage amount of each rod carrying barrel is 1-3 embryos according to the transplanting requirement, the number of the rod carrying barrels for carrying the rod carrying barrels is 1-4, namely when one patient needs to freeze and store about 8 embryos or less, one rod carrying barrel frame needs to be occupied, and when more than 8 embryos need to be occupied, 2 or more rod carrying barrel frames need to be occupied. The carrier system can facilitate the loading during freezing and also facilitate the rapid finding of the carrier bar frame from the liquid nitrogen tank and the extraction of the carrier bar from the carrier bar barrel clamped by the carrier bar frame during thawing. However, while such systems are convenient to use, they take up a lot of space. Each bucket can only put 26-28 loading rod barrel frames according to a liquid nitrogen tank of 47 liters and 10 buckets, and embryos of about 28 patients can be frozen in each bucket at most according to the calculation that each patient needs at least one loading rod barrel frame. A 47 liter 10-bucket liquid nitrogen tank can hold up to about 280 patient embryos. For a larger assisted reproductive center, if the number of egg taking cycles exceeds 500 per month on average, even if the space for unfreezing embryos is considered, the liquid nitrogen tank required by newly added frozen embryos can be increased by one month on average, and the extra space is occupied in a few years. This has limited the development of an assisted reproductive mechanism. Therefore, the space utilization efficiency of the existing micro biological sample freezing device and the corresponding loading technology thereof is relatively low, and the large-scale sample freezing storage and the development thereof cannot be met.

On the other hand, when freezing or thawing operation, the aluminum alloy material of the carrier rod barrel holder in the liquid nitrogen volatile vapor fog is easier to condense water vapor in the air and freeze than other plastic materials, which often causes the label part to be covered by condensed ice after exposing the liquid nitrogen for a certain time, thus causing inconvenience in operation.

Disclosure of Invention

In order to overcome the defects of the existing trace biological sample freezing device and the corresponding loading technology in the aspect of storage space utilization efficiency, the invention provides a micro carrier system for biological material freezing and a freezing storage technology thereof, so that the space occupied by the carrier device can be remarkably reduced while the operations of freezing loading, marking, storing, distinguishing, taking out, unfreezing and the like are simple and convenient, and the storage capacity of each liquid nitrogen tank (the number of cases for storing samples in the case of embryo vitrification freezing for assisted reproduction) is increased by about 3-4 times.

The invention provides a micro carrier system for cryopreservation of biological materials, which comprises a cryopreservation unit, a freezing unit and a freezing unit, wherein the cryopreservation unit comprises a carrying rod, a carrying rod sleeve and a carrying rod barrel; wherein,

the carrying rod sleeve is of a hollow structure, and at least one end of the carrying rod sleeve is provided with an opening communicated with the hollow structure;

the loading rod comprises a rod body and a freezing part connected with the bottom end of the rod body, a sleeving part is arranged between the rod body of the loading rod and the freezing part, a certain distance is reserved between the outer surface of the sleeving part at the position of the rod body of the loading rod and the outer surface of the rod body of the loading rod, the distance is equal to the wall thickness of an opening of the loading rod sleeve, the freezing part is inserted into a hollow structure of the loading rod sleeve to form a loading unit, and the loading rod sleeve is fixedly sleeved on the sleeving part of the loading rod;

the rod carrying cylinder is a hollow pipe with at least a closed bottom end and an average outer diameter less than 8.5mm (the average outer diameter refers to the average value of the maximum outer diameter and the minimum outer diameter, preferably 4-8.5mm, and more preferably 5-7 mm), and a rod loading opening capable of placing and taking out the loading unit from the rod carrying cylinder is formed in the top end or the side wall of the rod carrying cylinder.

Wherein, there may be a clearance between the inner wall of the rod carrying cylinder and the loading unit, or the inner wall of the rod carrying cylinder is contacted with the loading unit, and there is no clearance between the two, but the loading unit can slide in the rod carrying cylinder.

Wherein, the socket joint portion can be a carrying rod body or a part of the freezing storage piece.

The sleeve portion preferably continues to gradually decrease toward the cryopreserved section, and more preferably slightly gradually decreases toward the cryopreserved section.

In a preferred embodiment, the rod loading port is at least one notch arranged on the side wall of the rod carrying cylinder and used for placing and taking a loading unit in and out of the rod carrying cylinder, and the length of the loading unit is greater than the length from the bottom end of the notch to the bottom of the rod carrying cylinder but less than the length from the middle position of the notch to the bottom of the rod carrying cylinder.

Wherein the notch is preferably elongated.

Wherein a length of the slot may be greater than or equal to a length of the loading unit; and may even be less than the length of the loading unit and enable the loading unit in the slide holder to be removed from the slide holder through the slot.

Wherein the length of the loading unit is preferably less than the length from the 1/3 (more preferably 1/4, more preferably 1/5, more preferably 1/6, more preferably 1/7, more preferably 1/8) notch position from the bottom end of the notch to the bottom of the carrier rod barrel.

Preferably, the lower section of the slot is a throat section with a reduced opening width, and the length of the loading unit is greater than the length from the bottom end of the throat section to the bottom of the rod carrier, but less than the length from the top end of the throat section to the bottom of the rod carrier.

Preferably, the freezing and storing unit further comprises a blocking rod, and the blocking rod slides at a position above the bottom end of the notch of the rod carrying cylinder in the rod carrying cylinder, so that the blocking length of the notch is changed, and the rod loading port is opened or closed. The gear rod can be completely or only the lower section of the gear rod is positioned in the rod carrying cylinder.

Preferably, the top end of the rod carrying cylinder is provided with an opening, the lower section of the stop lever is inserted into the rod carrying cylinder from the opening at the top end of the rod carrying cylinder, and the upper section of the stop lever is thickened, so that the outer diameter of the upper section of the stop lever exceeds the inner diameter of the opening at the top end of the rod carrying cylinder to fix the stop lever and position the maximum shielding position of the stop lever on the notch.

Preferably, the stop lever is a hard straight hollow pipe, the bottom end of the stop lever is a closed end when the stop lever is completely positioned in the rod carrying cylinder, and the two ends of the stop lever are closed ends when only the lower section of the stop lever is positioned in the rod carrying cylinder; the top surface or the peripheral wall surface of the closed end is provided with small ventilation holes.

Preferably, a metal weight is arranged at the bottom end of the rod carrying cylinder.

In another preferred embodiment, the rod carrying cylinder rod loading port is a rod carrying cylinder port arranged at the top end of the rod carrying cylinder; the cryopreservation unit also comprises a sleeve, the sleeve is a long pipe with the upper end closed and the lower end open, the top and/or the peripheral wall of the upper end of the sleeve is provided with a vent hole, the lower section of the sleeve is provided with a connecting section which can be inserted into the rod carrying cylinder from the opening of the rod carrying cylinder, the outer diameter of the connecting section is gradually increased from bottom to top, and the maximum outer diameter of the connecting section is larger than or equal to (preferably equal to) the inner diameter of the opening of the rod carrying cylinder, so that the sleeve can be inserted into the opening of the rod carrying cylinder for fixation, the rod loading opening is closed, and the cryopreservation unit; the distance from the lower end of the sleeve after being inserted and fixed to the bottom of the rod carrying cylinder is greater than the length of the loading unit; the bottom of the rod carrying cylinder is provided with an iron block which can slide in the rod carrying cylinder.

Wherein a gap exists between the inner wall of the rod carrier and the lower end of the connecting section of the sleeve, and the gap is not enough for inserting the loading unit.

There may be a gap between the inner wall of the rod carrier and the iron block, and the gap is insufficient for insertion of a loading unit; or the inner wall of the rod carrying cylinder is contacted with the iron block, no gap exists between the inner wall of the rod carrying cylinder and the iron block, but the iron block can slide in the rod carrying cylinder.

When the rod loading port of the rod loading cylinder is used for unfreezing the freezing storage unit arranged at the top end of the rod loading cylinder, the miniature carrier system further comprises a magnetic positioning concave seat, a groove capable of accommodating the rod loading cylinder is arranged on the magnetic positioning concave seat, and a magnet block is arranged on the side wall of the groove at a position which is at least 3mm (preferably at least 6mm, and more preferably at least 9 mm) away from the bottom of the groove.

In another preferred embodiment, the freezing part of the carrier rod is a fine hollow tube structure or a strip structure.

Wherein the outer diameter of the fine hollow pipe structure is 0.3-3mm, more preferably 0.8-2.5mm, and more preferably 1.2-2.2 mm.

Wherein the inner diameter of the micro hollow tube is 0.1-2.8mm, more preferably 0.5-2.2mm, and more preferably 0.8-1.8 mm.

The cross-section of the strip-like structure is preferably concave.

Wherein the strip-like structure is a groove-shaped or arc-shaped piece and forms an end at the end. The ends may be pointed or rounded.

In any of the carrier systems of the present invention, the cross section of the carrier rod cylinder may be in various shapes including a circle, such as a circle, an ellipse, a semicircle, a semi-ellipse, a rectangle, a polygon, and even an irregular shape, such as a triangle, a quadrangle, a pentagon, a hexagon, etc.; but is preferably circular or oval or oblong, most preferably circular.

In any of the above carrier systems of the present invention, the part of the carrier rod other than the cryopreserving sheet may be a solid or hollow shape such as a cylinder, a partial cylinder, a prism, an elliptic cylinder, etc., and is preferably a cylinder.

In any of the above carrier systems of the present invention, the carrier sleeve may be cylindrical, prismatic, elliptical, etc., and is preferably cylindrical.

Preferably, the rod body of the carrying rod is tubular.

In another preferred embodiment, a label attaching portion is provided at a tip end of the rod body of the carrier rod, and the label attaching portion is a thin rod thinner than the rod body or a flat body narrower than the rod body.

The pore passages such as the vent holes, the small vent holes and the liquid through holes at the upper ends or the tops of the tube cavities have the functions of liquid nitrogen through flow and liquid nitrogen steam discharge, and can enter liquid nitrogen during freezing to form freezing protection or discharge liquid nitrogen steam pressure during thawing to prevent explosion.

In another preferred embodiment, 1 or more loading units are loaded in the rod carrier. The loading rod carrier with different inner diameters and shapes can be designed according to requirements so as to be suitable for loading different loading units or combined loading.

In another preferred embodiment, the carrier system further comprises a carrier rod sleeve stacking aid slot; the rod carrier sleeve stacking aid slot at least comprises three wall surfaces.

Preferably, the carrying rod stacking auxiliary slot comprises two clamping walls and a bottom wall, and the three wall surfaces are connected in pairs to form a triangular conical slot with an opening on one surface.

And, preferably, the dihedral angle formed between the two chuck walls may be a sharp corner or a rounded corner, and the circular arc radius of the rounded corner is smaller than that of the freezing storage slice groove.

Wherein, the carrier bar sleeve stacking auxiliary slot is preferably in a polygonal cone shape, such as a triangular cone, a quadrangular cone and the like, and is most preferably in a triangular cone shape.

In another preferred embodiment, the carrier system further comprises a bucket for placing the cryopreservation unit.

The second aspect of the invention provides a side-leakage type bucket suitable for storing a micro cryopreservation unit, wherein the bottom of the bucket is closed, and the side wall of the bucket is provided with a liquid through hole at least at a part which is 3mm higher than the bottom (preferably 5mm higher than the bottom, more preferably 7mm higher than the bottom, and more preferably 10mm higher than the bottom).

Preferably, the interior of the bucket is provided with a plurality of partition plates perpendicular to the bottom of the bucket, and the interior of the bucket is divided into a plurality of spaces (for example, 3-5 spaces) so as to reduce the number of frozen units in each space, thereby improving the speed of searching for the specific mark.

Preferably, a column seat is arranged in the bucket and comprises a base arranged at the bottom of the bucket and a column vertically arranged on the base.

Preferably, a circle of flat sunken bottom is arranged on the upright post seat around the upright post, the periphery of the sunken bottom is an outer ring bottom with a raised plane, a plurality of bottom edge clamping grooves allowing the bottom edges of the separating sheets to be clamped in are radially arranged on the outer ring bottom by taking the upright post as a center, the bottoms of the clamping grooves are parallel to the sunken bottom, and the depth of the clamping grooves, namely the height difference between the outer ring bottom and the sunken bottom, is at least 1 mm. The design can make the bottom edge slot in the bucket many. When the quantity of the freezing units in the original interval is reduced greatly due to continuous unfreezing, the separating sheet can be withdrawn from the clamping groove and can be moved to a new clamping groove in the bucket to be inserted. This allows compression of the compartment without affecting the separation of the freezing units in the original compartment, leaving a new space for a new separation.

Preferably, a plurality of side clamping grooves which are perpendicular to the base and allow a certain side of the separating sheet to be clamped in are also arranged on the upright column.

Preferably, the base (the plane bottom) is provided with a plurality of bottom edge clamping grooves which are communicated with the side edge clamping grooves in a one-to-one correspondence manner and allow the bottom edges of the separation sheets to be clamped in.

Preferably, the upright post is a central upright post, and the length of the top edge of the separating sheet is equal to the distance from the bottom of the side clamping groove to the inner wall of the bucket, so that the fixing effect is further better generated.

Preferably, the width of the lower part of the separating sheet is reduced from the side edge which is not inserted into the side clamping groove (the side edge of the separating sheet inserted into the side clamping groove is kept to be a neat edge and is perpendicular to the bottom edge), and the reduced width is at least larger than the depth of the side clamping groove. When the inserted separating sheet is withdrawn upwards to the reduced side edge and exposed out of the edge of the hanging bucket, the separating sheet can be taken out from the side edge clamping groove towards the lateral outer direction, the separating sheet is moved to another clamping groove in the hanging bucket, then the separating sheet is laterally clamped into the side edge clamping groove and then downwards inserted into the bottom edge clamping groove. When the quantity of the freezing units in the original interval is reduced greatly due to continuous unfreezing, the operation of moving the separating sheet in the bucket can realize the compression of the interval without influencing the separation of the freezing units in the original interval, and a new space is reserved for new separation.

Preferably, the upright columns can also be eccentric upright columns, and the width of each partition sheet is the length linearly extending from the bottoms of the clamping grooves at different sides to the inner wall of the bucket along the clamping groove at the bottom side, so that the widths are not uniform.

The separator sheets may be of different colours.

The third aspect of the present invention provides a cryopreservation method for biological materials, which comprises the following steps:

step 1, providing the micro carrier system, placing biological materials in a freezing part for freezing in liquid nitrogen, sleeving a rod carrying sleeve to form a loading unit, placing the loading unit into a rod carrying cylinder from a rod loading port of the rod carrying cylinder, and closing the rod loading port to form a freezing unit;

step 2, providing any one of the lifting buckets, dividing the inner space of the lifting bucket into a plurality of sections, and placing the lifting bucket in a liquid nitrogen tank;

and 3, filling the freezing unit obtained in the step 1 into the bucket in the liquid nitrogen tank provided in the step 2 for freezing. In a preferred embodiment, the sleeve carrier sleeve in step 1 comprises:

step 1.1, providing a loading rod sleeve stacking auxiliary slot; the loading rod sleeve stacking auxiliary slot at least comprises three directly contacted wall surfaces which enclose two clamping walls and a bottom wall and are intersected to form a bottom tricone angle; placing the bottom three-cone angle at the bottommost position to enable the connecting edge between the two clamping walls to be obliquely lifted; placing the rod-carrying sleeves in the rod-carrying sleeve stacking slot in a single or multiple manner, so that the bottom ends of the rod-carrying sleeves are abutted against the bottom wall; the carrying rod sleeve is under the action of gravity and the blocking of the two clamping walls, and the carrying rod sleeve at the lowest part is naturally positioned in the middle of the connecting edge of the two clamping walls;

and step 1.2, the front end of the carrying rod is tightly attached to the two wall surfaces of the two clamping walls of the two clamping wall connecting edges of the carrying rod sleeve stacking auxiliary slot, the freezing part can automatically align with the carrying rod sleeve inlet in the middle position of the two clamping wall connecting edges, and the carrying rod moves towards the carrying rod sleeve and is inserted into the carrying rod sleeve.

The invention abandons the rod carrying cylinder bracket occupying the most space in the traditional carrier system, and can make the diameter of the rod carrying cylinder smaller because the thickness of the rod carrying cylinder sleeved with the rod carrying sleeve is reduced, and the rod carrying cylinder is designed into various sizes and shapes to adapt to various freezing storage requirements under the condition of the smallest occupation, so that the integral storage capacity of the liquid nitrogen tank is enlarged to about 3-4 times of the original storage capacity. Meanwhile, the freezing and storing unit does not need an aluminum alloy carrying rod barrel frame, so that the inconvenience that the condensation ice is often fixed and covered on the label in the traditional technology is solved, and the sleeving operation of the carrying rod during freezing and storing is simpler and faster due to the design and application of the carrying rod sleeve stacking auxiliary slots.

The invention has the advantages that the space occupied by freezing the trace biological sample is reduced by times on the basis of not influencing or even increasing the use convenience, and the occupied area required by freezing storage is greatly saved. Is especially suitable for freezing and storing trace biological materials such as vitrified frozen embryos and the like.

Drawings

FIG. 1 is a schematic diagram of a conventional carrier system and loading process;

FIG. 2 is a schematic diagram of a microcarrier system and loading process according to an embodiment of the present invention;

FIG. 3 is a schematic view of a microcarrier system and loading and unloading process according to an embodiment of the present invention;

FIG. 4 is a schematic diagram of a prior art bucket and a bucket of the present invention, wherein A is the prior art bucket, the bottom surface of which is provided with a fluid hole, B is the bucket of the present invention, the side wall of which is provided with a fluid hole;

FIG. 5 is a schematic view of a bucket and an assembly process according to the present invention;

FIG. 6 is a schematic view of an exemplary embodiment of a stacking aid slot for a rod carrier and method of use;

FIG. 7 is a schematic diagram of a freezing section of the loading rod according to an embodiment of the present invention;

FIG. 8 is a schematic diagram of the structure of the connecting portion of the carrier rod and the freezing portion inserted into the carrier rod sleeve according to one embodiment of the present invention

FIG. 9 is a schematic diagram of a groove structure of the freezing section according to an embodiment of the present invention;

FIG. 10 is a schematic view of the groove-shaped cryopreservation section of FIG. 9 inserted into a rod carrying sleeve using the rod carrying sleeve insertion and stacking slot of FIG. 6;

figure 11 is a cross-sectional view of a different shape of carrier rod barrel for multiple loading units.

Detailed Description

The invention is described in detail below with reference to specific embodiments in conjunction with the accompanying drawings.

Example 1

In the embodiment, the original rod carrying cylinder 3' with an opening at one end is changed into a thin and lengthened rod carrying cylinder 3 which is provided with a long strip-shaped window, namely a notch 31 (rod loading port) from the side surface and at least has a closed bottom end; the taking and placing of the carrier rod in the carrier rod cylinder are carried out from the side notch 31 through tweezers; abandoning the external force clamping type pole carrying cylinder cap 4', changing the lower section into a stop lever 4 which is inserted into the pole carrying cylinder from the top end opening of the pole carrying cylinder and is fixed by friction, controlling the open length of the notch 31 by the occupying length of the stop lever at the notch 31, carrying out the picking and placing operation of the pole carrying 1 (loading unit) when the notch is open longer, and stopping the sliding of the pole carrying 1 in the pole carrying cylinder 3 when the notch is open shorter. Therefore, an external rod carrying cylinder cap is not needed, and the rod carrying cylinder frame occupying the most space and used for clamping the rod carrying cylinder cap is abandoned correspondingly. Meanwhile, the carrying rod barrel also plays a role of an independent cryopreservation unit with different biological sample source marks, which is played by the original carrying rod barrel frame, namely, the upper end of the carrying rod barrel is marked, and the carrying rod barrel can be identified and taken and placed conveniently like the mark on the top end of the original carrying rod barrel frame to form the independent cryopreservation unit. The barrel is multipurpose, and a large amount of space required by a barrel peripheral matching device is saved under the condition of not influencing the original operation simplicity. Moreover, because the new freezing unit carrier rod cylinder is made of plastic, the low thermal conductivity is realized, the freezing of water vapor in air due to the fact that the label part is exposed out of liquid nitrogen for a certain time can be avoided, and the inconvenience that the label is often covered by the existing aluminum alloy carrier rod cylinder frame due to the condensation ice is solved.

Referring to fig. 2, the carrier system provided in this embodiment mainly comprises a carrier rod 1, a carrier rod sleeve 2, a carrier rod cylinder 3, and a stopper rod 4, wherein the bottom end (freezing part 12) of the carrier rod is inserted into the carrier rod sleeve 2, and then the carrier rod and the stopper rod are placed into the carrier rod cylinder 3, and the lower section of the stopper rod 4 is inserted into the carrier rod cylinder 3 from the top opening of the carrier rod cylinder 3.

The rod carrying sleeve 2 is a hollow pipe with an open upper end and a closed lower end.

The carrying rod 1 comprises a rod body 11 and a freezing part 12 connected with the bottom end of the rod body 11, and the freezing part 12 is inserted into the carrying rod sleeve 2 to form a loading unit.

The rod carrying cylinder 3 is an empty pipe with a closed bottom end and an open upper end with an outer diameter of 7mm (the pipe can be designed into other outer diameters according to actual requirements), and the side wall of the rod carrying cylinder 3 is provided with a long-strip-shaped notch 31 which can put the loading unit into and take the loading unit out of the rod carrying cylinder 3.

The lower section of the slot 31 is a throat section 311 with a reduced opening, and the length of the loading unit is greater than the length from the bottom end of the throat section 311 to the bottom of the rod carrier 3, but less than the length from the top end of the throat section 311 to the bottom of the rod carrier 3.

The top end of the rod carrying cylinder 3 is opened, the lower section of the baffle rod 4 is inserted into the rod carrying cylinder 3 from the top end opening of the rod carrying cylinder 3 and slides at a position above the bottom end of a rod carrying cylinder notch 31 in the rod carrying cylinder 3, so that the shielding length of the notch 31 is changed, and a rod loading port (namely the notch 31) is opened or closed; the upper section of the stop lever 4 is thickened, so that the outer diameter of the upper section of the stop lever 4 exceeds the inner diameter of the opening at the top end of the rod barrel 3, the stop lever 4 is fixed, and the shielding position of the stop lever 4 on the notch 31 is positioned.

The stop lever 4 is a hard straight hollow tube, two ends of the stop lever are closed ends, and small ventilation holes are formed in the top surface or the peripheral wall surface of each closed end. When the freezing unit is used, the hollow pipe is filled with liquid nitrogen to remove buoyancy caused by air, so that the sinking is facilitated, and meanwhile, the more liquid nitrogen filled, the more liquid.

The tip of the rod body 11 of the carrier rod is provided with a label attaching portion 13, and the label attaching portion 13 is a thin rod thinner than the rod body or a flat body narrower than the rod body.

Referring to fig. 7 and 8, a sleeving part 14 is arranged between the rod carrying body 11 and the freezing part 12, a certain distance is reserved between the outer surface of the sleeving part 14 at the position of the rod carrying body 11 and the outer surface of the rod carrying body 11, and the distance is equal to the wall thickness of the opening of the rod carrying sleeve 2, so that the rod carrying sleeve 2 and the rod carrying rod 1 form a straight rod shape without outward protrusion after the rod carrying sleeve 1 is sleeved with the rod carrying rod 1, and the space occupied by the rod carrying sleeve due to the fact that the rod carrying sleeve is wrapped around the periphery of the rod carrying rod is.

However, this inevitably results in the freezing end of the rod-carrying sample being reduced and the opening of the rod-carrying sleeve being reduced, which may cause difficulty in visual positioning when the top of the freezing end of the rod-carrying sample is inserted into the opening of the rod-carrying sleeve. To this end, non-visual automatic positioning insertion devices and methods are further devised to overcome:

the freezing part 12, which is one end of the slide for freezing the biological sample, is maintained at a certain diameter, and if the freezing part is a freezing piece of a strip structure, the freezing piece is formed in a groove shape as shown in fig. 7, and the tip 15 is a pointed shape. While the carrier system of this embodiment includes a rod carrier stacking slot, fig. 6 is a schematic view of the structure, components and operation method of the rod carrier stacking slot according to a preferred embodiment of the present invention. As shown in fig. 7 and 8, one end of the frozen sample of the slide bar 1 includes a sleeve portion 14, a frozen portion 12, and a frozen portion end portion 15. The outer diameter of the sleeving part 14 is gradually reduced towards the direction of the freezing part, so that the freezing part is conveniently and gradually sleeved with the carrier rod sleeve 2, and the carrier rod 1 and the carrier rod sleeve 2 form straight butt joint due to the same outer diameter after being sleeved tightly without convex; the freezing part 12 of the carrying rod 1 is a long strip groove shape, the top end is a sharp end part 15 formed by cutting the groove, and micro biological samples such as 1-3 embryos are placed in the groove close to the rear part of the end part 15 of the freezing part. This groove configuration can shield frozen embryos from the risk of being rubbed during handling. Meanwhile, as shown in fig. 6, a triangular-cone-shaped rod-carrying sleeve stacking slot-aid 6 is provided, which comprises two clamping walls 61 and a bottom surface 62, wherein a groove shape with one open surface is formed by the enclosing, and two wall surfaces 61 are spliced to form dihedral angles. When the carrier rod case stacking aid inserting groove 6 is obliquely arranged as shown in fig. 6, a plurality of carrier rod cases 2 stacked therein can be automatically and neatly positioned at the bottom corner by gravity, wherein the bottommost carrier rod case 2 is always flatly attached to two side walls 61 of the dihedral angle, and when the carrier rod 1 is required to be inserted into the carrier rod case 2 after being filled with a biological sample, as shown in fig. 6 and 10, the outer wall of the carrier rod body 11 can be flatly attached to both side walls 61 of the dihedral angle of the carrier rod case stacking aid inserting groove 6, or the concave portion of the carrier rod front end sheet strip freezing storage portion 12 can be inserted toward the carrier rod case 2 while the freezing storage portion tip 15 at the top end thereof is automatically lifted and suspended due to the concave shape of the freezing storage portion 12 without contacting both side walls 61 of the carrier rod case stacking aid inserting groove 6, and the shape and the dimension thereof are designed such that the freezing storage portion tip 15 can be automatically aligned with the opening of the carrier rod case 2, this ensures that the slimmed cryopreservation portion 12 can still be inserted into the rod sleeve 2 more conveniently without careful observation.

In the prior art, the thickest structure of the freezing storage unit is a rod carrying barrel frame, namely an aluminum alloy frame, the specification and the shape of the aluminum alloy frame are the same, so that even one rod carrying rod needs to occupy one freezing storage unit as 4 rod carrying rods, and 5 rod carrying rods need to use 2 freezing storage units which are twice as many as 4 rod carrying rods. Both of these loads present a waste of space. In this embodiment, the carrier rod barrel is made of plastic, so that different specifications and shapes can be made according to actual loading capacity without uniform specification like an aluminum alloy frame, and space is reasonably occupied. As most of 6-8 embryos need 3-4 carrying rods, carrying rod barrels with the loading capacity of 3 carrying rods as the minimum specification can be designed, carrying rod barrels with the loading capacities of 4 and 5 are additionally designed, and different cross section areas and shapes of the carrying rod barrels are designed according to different loading capacities, and besides the traditional round shape, the carrying rod barrels also comprise an oval shape, a sector shape, a square shape and the like. Thus, 1-3 can be arranged in 3 standard cylinders, 4-5 can be respectively arranged in 4 and 5 standard cylinders, 6 and more can be arranged in a larger design, or a plurality of rod carrying cylinders with the above specification are combined for loading, such as 6 rods are divided into 2 rods for 3 rods, 7 rods are divided into 1 rod for 3 rods and 1 rod for 4 rods, and the space is fully utilized; it is also possible to use a single pack or 2 pack scheme which, while taking up slightly more space when frozen, can speed up the vacation of space when thawed. For example, a single loading solution, i.e. a carrier rod cartridge designed to be the smallest and accommodating only one loading unit (carrier rod and carrier rod sleeve). This will allow each removed freezing unit to be emptied after each thaw, eliminating the need to replace the freezing unit to the liquid nitrogen tank, simplifying the thawing process and speeding up the evacuation of space.

FIG. 11 is a schematic diagram of the cross-sectional shape design components of the carrier rod barrel of the carrier system of the present invention. The carrier rod barrel 4 can accommodate a plurality of loading units, can be made of plastic, preferably transparent or translucent plastic, and can be dimensioned differently according to the actual loading: as shown in fig. 11 a, a rod carrier 4 with 3 loading units as the maximum loading capacity is designed, and the 3 loading units therein will automatically be in the minimum occupying state of the triangular position, so that a circular cross-section design is adopted; as shown in fig. 11B, 4 loading units are designed to be the rod carrying cylinder 3 with the maximum loading capacity, and the 4 loading units therein will automatically be in the minimum occupying state of the diamond position, so that an oval cross-section design can be adopted; as shown in fig. 11C, 5 loading units are designed as the rod carrying cylinder 3 with the maximum loading capacity, and the 5 loading units therein will automatically be in the minimum occupying state of the pentagonal position, so that the circular cross-section design can be adopted; various other carrier rod cross-sectional shapes can be designed, including scallops, squares, and the like. Thus, 1 to 3 rod carrying cylinders 3 with the specification of 3 can be arranged, 4 to 5 rod carrying cylinders 3 with the specification of 4 and 5 can be respectively arranged, 6 or more rod carrying cylinders with the specification can be arranged by a larger design or a plurality of rod carrying cylinders with the specification can be combined, and the space is fully utilized. Meanwhile, the space of the original bucket of one unit can be divided into a plurality of spaces (such as 3-5 spaces) so as to reduce the number of the frozen units in each space, thereby improving the speed of searching for the specific mark.

Example 2

In the embodiment, the original rod carrying cylinder 3' with an opening at one end is changed into a thin and lengthened rod carrying cylinder 3 which is opened from the top end, namely a cylinder opening of the rod carrying cylinder, is closed at the bottom end, and is provided with a movable iron block at the bottom in the cylinder; the taking-out of the loading rod in the loading rod cylinder is as follows: the movable iron block is driven to move upwards through the magnetic positioning concave seat, and the loading rod is lifted to be exposed out of the barrel opening of the loading rod at the top end; abandoning the external force clamping type rod carrying cylinder cap 4', changing into a sleeve 5 which has a lower connecting section and can be inserted into the rod carrying cylinder from the opening of the rod carrying cylinder and fixed by friction so as to close the rod loading opening and prolong the length of the freezing storage unit to exceed the depth of the bucket of the liquid nitrogen tank bucket. Thus, the most space-consuming rod carrier frame for clamping the rod carrier cap is abandoned. Meanwhile, the upper end of the sleeve is marked, and the identification and the taking and placing can be conveniently carried out like the marking of the top end of the original carrying rod barrel frame. Thus, under the condition of not influencing the original operation simplicity, a large amount of space required by a barrel periphery matching device is saved. Moreover, because the sleeve 5 is made of plastic and has low thermal conductivity, when the sleeve is obliquely placed in the liquid nitrogen box, the cylinder wall of the part exposed out of the liquid nitrogen cannot be greatly influenced by low-temperature conduction of the part in the liquid nitrogen, namely, the cylinder wall cannot have low temperature like an aluminum alloy rod-carrying cylinder frame, and therefore the label part cannot be solidified and frozen due to excessive condensation of water vapor in air caused by exposure of the liquid nitrogen for a certain time. This has solved the inconvenience that present aluminum alloy carries pole bobbin bracket often covers the label because of the condensation ice.

Referring to fig. 3, the carrier system provided in this embodiment mainly includes a carrier rod 1, a carrier rod sleeve 2, a carrier rod barrel 3, and a sleeve 5, wherein the bottom end (the freezing portion 12) of the carrier rod 1 is inserted into the carrier rod sleeve 2, and then the carrier rod 1 and the carrier rod barrel 3 are loaded together from the port of the carrier rod barrel at the top end of the carrier rod barrel 3, and the lower section of the sleeve 5 is inserted into the carrier rod barrel 3 from the port of the carrier rod barrel at the top end of the carrier rod barrel 3.

The rod carrier sleeve 1 and the rod carrier 2 in the embodiment are the same as the rod carrier sleeve 1 and the rod carrier 2 in the embodiment 1.

The rod carrying cylinder 3 of this embodiment is an empty tube having a closed bottom end and an open upper end, and has an outer diameter of 6mm, and the open upper end of the rod carrying cylinder constitutes a rod carrying cylinder port (i.e., a rod loading port) through which the loading unit can be loaded into and unloaded from the rod carrying cylinder.

The sleeve 5 is a long pipe with the upper end closed and the lower end opened, the lower section of the sleeve 5 is provided with a connecting section 51 inserted into the rod carrying cylinder from the cylinder opening of the rod carrying cylinder, the outer diameter of the connecting section 51 is gradually increased from bottom to top, and the maximum outer diameter of the connecting section 51 is equal to the inner diameter of the cylinder opening of the rod carrying cylinder, so that the sleeve 5 can be inserted into the cylinder opening of the rod carrying cylinder for fixation, so as to close the rod loading opening and prolong the freezing unit; the distance from the lower end of the inserted and fixed sleeve 5 to the bottom of the rod carrying cylinder 3 is larger than the length of the loading unit.

The bottom of the rod carrying cylinder 3 is provided with an iron block 32 which can slide in the rod carrying cylinder 3.

There may be a gap between the inner wall of the carrier rod barrel and the iron block 32, and the gap is not sufficient for insertion of a loading unit; or the inner wall of the rod carrier cylinder is in contact with the iron block 32, and no gap exists between the two, but the iron block can slide in the rod carrier cylinder.

When the loading unit needs to be taken out of the rod carrying cylinder, the iron block 32 in the rod carrying cylinder can be attracted by the magnet to slide upwards, and the loading unit is driven to be exposed out of the opening of the rod carrying cylinder.

In a preferred embodiment, the microcarrier system further comprises a magnetic positioning recess 7, wherein a groove capable of accommodating the carrier rod barrel is arranged on the magnetic positioning recess, and a magnet block is arranged on the side wall of the groove at a position which is at least more than 3mm away from the bottom of the groove. When the loading unit needs to be taken out of the rod carrying cylinder, the rod carrying cylinder 3 is placed in the groove of the magnetic positioning concave seat, and the magnet block on the side wall of the groove can cause the iron block 32 in the rod carrying cylinder to move upwards to the position of the magnet to drive the rod carrying cylinder to be exposed out of the opening of the rod carrying cylinder.

The carrier rod barrel of the present embodiment can also accommodate a plurality of loading units, can be made of plastic, and can be made into different specifications according to the actual loading capacity, as shown in fig. 11.

In the above embodiment 1 or 2, the freezing part of the carrier rod includes the freezing sheet of the strip structure or the freezing microtube of the fine hollow tube structure. In the case of a fine hollow tube, the outer diameter is 0.3-3mm, preferably 1mm, and the inner diameter is 0.8mm, and when the tube is frozen for use, a trace amount of biological sample such as 1-3 embryos is sucked and placed into the fine hollow tube.

Example 3

The micro carrier system provided by the invention has the advantages that the carrying rod cylinder is thin, the liquid nitrogen amount in the carrying rod cylinder is small when the micro carrier system is frozen, and the liquid nitrogen in the bucket is quickly leaked from the bottom mesh when the bucket is extracted. The small amount of liquid nitrogen in the rod carrying cylinder is in a non-liquid nitrogen environment and is easy to completely volatilize, and at the moment, the biological material cryopreservation part is probably not in the protection of the liquid nitrogen, so that the biological material is easy to abnormally thaw and is easy to fail in cryopreservation.

Referring to fig. 4 and 5, the conventional bucket 8 'is closed at its side wall and has a mesh-shaped liquid through hole 81' at its bottom, and this embodiment provides a side-leakage-type bucket 8 suitable for storing a micro-freezing unit, the bottom of the bucket is closed, and the liquid through hole 81 is formed at a portion of the side wall of the bucket which is at least 3mm higher than the bottom. When the bucket is raised to be higher than the liquid nitrogen surface of the liquid nitrogen tank, the liquid nitrogen with the non-leakage height at the bottom of the bucket can delay the volatilization time of a small amount of liquid nitrogen in the loading rod cylinder.

In a preferred embodiment, the interior of the bucket is provided with a plurality of dividing plates 91 perpendicular to the bottom of the bucket to divide the interior of the bucket into a plurality of spaces (e.g., 3-5) to reduce the number of frozen units in each space, thereby increasing the speed of finding a particular mark.

In a further preferred embodiment, a column base 9 is arranged in the bucket, the column base 9 includes a base 92 arranged at the bottom of the bucket and a column 93 vertically arranged on the base 92, wherein a plurality of side slots 94 allowing a certain side of the partition sheet 91 to be clamped in are arranged on the column 93, the side slots 94 allowing the partition sheet 91 to be clamped in are perpendicular to the base, and a plurality of bottom slots 95 allowing the partition sheet 91 to be clamped in are arranged on the base 92, and are in one-to-one correspondence with the side slots 94.

The length of the top edge of the partition plate 91 is equal to the distance from the bottom of the side locking groove 94 to the inner wall of the bucket 8, thereby further improving the fixing effect.

The width of the lower portion of the separator 91 is reduced from the side edge that is not inserted into the side edge notch 94 (keeping the side edge of the separator inserted into the side mark notch perpendicular to the bottom edge), by at least a greater depth than the side edge notch 94. This allows the inserted separator 91 to be withdrawn upward until the reduced side edge is exposed to the rim of the bucket, and then the separator 91 can be removed from the side edge slot 94 in the lateral outward direction, moved within the bucket to another slot unit (consisting of a side edge slot and a bottom edge slot that are in communication), then laterally clipped into the side edge slot 94, and then inserted downward into the bottom edge slot 95. When the quantity of the freezing units in the original interval is reduced greatly due to continuous unfreezing, the operation of moving the separating sheet in the bucket can realize the compression of the interval without influencing the separation of the freezing units in the original interval, and a new space is reserved for new separation.

When the biological material is frozen, the micro carrier system provided in embodiment 1 or embodiment 2 can be adopted, the biological material is placed in the freezing part and frozen in liquid nitrogen, the rod carrying sleeve is sleeved to form a loading unit, the loading unit is placed into the rod carrying cylinder from the rod carrying port of the rod carrying cylinder, and the rod carrying port is closed to form the freezing unit; dividing the inner space of the bucket into a plurality of sections, and placing the bucket in a liquid nitrogen tank; and (4) putting the freezing unit into a bucket in a liquid nitrogen tank to finish freezing and loading.

The above examples 1-3 can be used as a revolutionary human embryo vitrification freezing carrier system to save a great deal of space for vitrification freezing of human embryos (or ova).

Example 4

When certain tissues such as ovary tissue slices, small cartilage slices, hair roots, tumor small blocks or other specific tissue small blocks need single or micro independent freezing storage in scientific research, detection or other application processes, the freezing storage by using a traditional conventional freezing storage tube with the capacity often causes that the tissues are difficult to find and lost after being thawed, the freezing storage recovery efficiency is low, and too much space is occupied during large-scale independent freezing storage. The human embryo vitrification freezing carrier system reformed by the invention can avoid the defect of excessive occupied space, solve the problem of difficult searching and loss after unfreezing due to small bearing range of the freezing part, and ensure good freezing effect and improved recovery survival and quality because the freezing and thawing speed of the freezing and thawing part is far faster than that of a freezing and thawing pipe with conventional capacity. The technical system and operation process are similar to those of the human embryo vitrification freezing carrier system and technology, and the embryo biological sample is changed into a specific tissue as described in the previous embodiments 1 and 2.

Example 5

When some cells such as human micro semen, testicular sperms, micro blood cells or other specific micro cells need micro independent cryopreservation in scientific research, detection or other application processes, the cryopreservation by using the traditional conventional capacity cryopreservation tube often causes that the cells are difficult to find and lost after thawing, the cryopreservation recovery efficiency is low, and excessive space is occupied during large-scale independent cryopreservation. The human embryo vitrification freezing carrier system reformed by the invention can avoid the defect of excessive occupied space, solve the problem of difficult searching and loss after unfreezing due to small bearing range of the freezing storage piece, and ensure good freezing storage effect and improved recovery survival and quality because the freezing and thawing temperature changing speed of the freezing storage piece is far faster than that of the freezing and thawing temperature changing speed of the freezing storage tube with the conventional capacity. The technical system and operation process are similar to those of the human embryo vitrification freezing carrier system and technology, and as described in the previous examples 1 and 2, the embryo is changed into a specific cell drop, but the thawing recovery is different.

The thawing of examples 1, 2 and 4 was carried out by inserting the freezing part 12 of the carrier rod 1 into a pre-warmed thawing solution, which is usually used to keep its temperature from being excessively affected by room temperature by a large amount of the solution. This makes the thawed micro cells diffuse in a large amount of fluid for the purpose of making the specific cell search difficult, for example, the number of sperms passing through the testis is often only several or several tens, and the cells cannot be collected in about 1 ml of fluid. In example 5, the freezing part 12 of the slide bar 1 was inserted into the preheated mineral oil in the culture dish, and after the frozen droplet on the freezing part was thawed, the droplet was inserted into another droplet of the culture medium under the mineral oil in the culture dish, so that the droplet of the sample and the cells thereof on the freezing part 12 were thawed into the new droplet of the culture medium, and then the new droplet was used for further microscopic thawing and use while searching for a trace amount of cells.

The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.

Claims

1. A micro carrier system for cryopreservation of biological materials is characterized by comprising a cryopreservation unit, wherein the cryopreservation unit comprises a carrying rod, a carrying rod sleeve and a carrying rod cylinder; wherein,

the rod carrying cylinder is a hollow pipe with at least a closed bottom end and an average outer diameter smaller than 8.5mm, and a rod loading port capable of placing and taking out the loading unit from the rod carrying cylinder is arranged at the top end or the side wall of the rod carrying cylinder;

the rod loading port of the rod carrying cylinder is a rod carrying cylinder port arranged at the top end of the rod carrying cylinder; the freezing and storing unit also comprises a sleeve, the sleeve is a long pipe with the upper end closed and the lower end opened, the top and/or the peripheral wall of the upper end of the sleeve is provided with a vent hole, the lower section of the sleeve is provided with a connecting section which can be inserted into the rod carrying cylinder from the cylinder opening of the rod carrying cylinder, the outer diameter of the connecting section is gradually increased from bottom to top, and the maximum outer diameter of the connecting section is larger than or equal to the inner diameter of the cylinder opening of the rod carrying cylinder, so that the sleeve can be inserted into the cylinder opening of the rod carrying cylinder for fixing, the rod loading opening is closed; the distance from the lower end of the sleeve after being inserted and fixed to the bottom of the rod carrying cylinder is greater than the length of the loading unit; the bottom of the rod carrying cylinder is provided with an iron block which can slide in the rod carrying cylinder;

the miniature carrier system also comprises a magnetic positioning concave seat, a groove capable of accommodating the rod carrying barrel is arranged on the magnetic positioning concave seat, and a magnet block is arranged on the side wall of the groove at a position which is at least 3mm away from the bottom of the groove.

2. The microcarrier system of claim 1, wherein the loading port is at least one slot in a sidewall of the carrier for allowing a loading unit to be loaded into and removed from the carrier, the loading unit having a length greater than the length of the slot from the bottom end of the slot to the bottom of the carrier but less than the length of the slot from the middle of the slot to the bottom of the carrier.

3. The microcarrier system of claim 2, wherein the lower slot section is a throat section with a reduced opening width, and the loading unit has a length that is greater than the length from the bottom of the throat section to the bottom of the stem carrier but less than the length from the top of the throat section to the bottom of the stem carrier.

4. The microcarrier system of claim 2, wherein the cryopreservation unit further comprises a stopper that slides in the interior of the carrier at a location above the bottom end of the slot of the carrier to change the length of the shield for the slot to open or close the loading port.

5. The microcarrier system of claim 4, wherein the top opening of the carrier roller has a lower stop bar section that is inserted into the carrier roller through the top opening of the carrier roller, and the upper stop bar section is thickened such that the outer diameter of the upper stop bar section exceeds the inner diameter of the top opening of the carrier roller to secure the stop bar and position the stop bar in a position that blocks the slot.

6. The microcarrier system of claim 4 or 5, wherein the stop is a rigid, hollow tube having at least a closed bottom end with a small vent hole in the top or peripheral wall of the closed end.

7. The microcarrier system of claim 1, wherein the cryopreservation section is a chip structure or a fine hollow tube structure with an outer diameter of 0.3-3 mm.

8. The microcarrier system of claim 7, wherein the cross-section of the ribbon structure is concave.

9. The microcarrier system of claim 1, wherein the carrier rod body is tubular.

10. The microcarrier system of claim 1, wherein the carrier rod has a label-bearing portion at the tip of the rod body, said label-bearing portion being a thin rod thinner than the rod body or a flat body narrower than the rod body.

11. The microcarrier system of claim 10, wherein 1 or more loading units are loaded in the carrier spool.

12. The microcarrier system of claim 1, further comprising a carrier bar nest stacking aid slot, wherein the carrier bar nest aid slot comprises two sandwiching walls and a bottom wall, and three walls are connected in pairs to form a triangular pyramid slot with an opening at one side.

13. The microcarrier system of claim 1, further comprising a bucket for holding the cryopreservation unit.

14. A side-leakage bucket adapted for storing a microcarrier system according to any of claims 1-13, wherein the bottom of the bucket is closed and the side wall of the bucket is provided with a liquid through hole at least 3mm higher than the bottom.

15. The bucket of claim 14 wherein the interior of the bucket has a plurality of dividers positioned perpendicular to the bottom of the bucket to divide the interior of the bucket into a plurality of spaces to reduce the number of cryopreserved cells in each space and thereby increase the speed of searching for a particular marked cryopreserved cell.

16. The bucket according to claim 15, wherein the bucket is internally provided with a column base, the column base comprises a base arranged at the bottom of the bucket and a column vertically arranged on the base, a circle of flat sunken bottom is arranged on the column base around the column, the periphery of the sunken bottom is an outer ring bottom which is raised in a plane, a plurality of bottom edge clamping grooves allowing the bottom edges of the separating sheets to be clamped are radially arranged on the outer ring bottom by taking the column as a center, and the bottoms of the clamping grooves are level with the sunken bottom.

17. The bucket of claim 15, wherein the bucket has a column base, the column base comprises a base disposed at the bottom of the bucket and a column vertically disposed on the base, and the column has a plurality of side slots perpendicular to the base for allowing a side of the partition to be inserted therein.

18. A method for cryopreserving biological materials, using the microsupport system of claim 1 and the bucket of any one of claims 14 to 17, comprising the steps of:

step 1, providing the micro-carrier system of claim 1, placing biological materials in a freezing part for freezing in liquid nitrogen, sleeving a rod carrying sleeve to form a loading unit, placing the loading unit into a rod carrying cylinder from a rod loading port of the rod carrying cylinder, and closing the rod loading port to form a freezing unit;

step 2, providing the bucket as claimed in any one of claims 14 to 17, wherein the inner space of the bucket is divided into a plurality of sections, and placing the bucket in a liquid nitrogen tank;

and 3, filling the freezing unit obtained in the step 1 into the bucket in the liquid nitrogen tank provided in the step 2 for freezing.