阅读说明：本技术 一种细胞冷冻辅助装置 (Cell freezing auxiliary device ) 是由 武旭临 于 2020-07-22 设计创作，主要内容包括：本发明属于生物材料冷冻技术领域,公开了一种细胞冷冻辅助装置。本发明包括贴合于载杆上端的容器皿,容器皿的中部设置有贯穿孔,贯穿孔的下端与载杆的生物材料装载部相对应；所述容器皿的上端设置有出液槽,容器皿的内部设置有排液腔,排液腔分别连通出液槽和贯穿孔。本发明便于细胞冷冻前对细胞的处理,便于快速精准的将细胞放入贯穿孔内,也便于后期细胞准确的落入载杆上,使包裹生物材料的冷冻液体积小而稳定,使得操作更加方便快捷,也不需要再移动细胞,节省了操作步骤；出液槽通过排液腔与贯穿孔相连,方便将各种玻璃化溶液吸走,同样可以使得用过的玻璃化溶液的处理更加轻松方便,效率也更高,整体保证了操作的稳定性、及时性和安全性。(The invention belongs to the technical field of biological material freezing, and discloses a cell freezing auxiliary device. The invention comprises a container vessel which is attached to the upper end of a carrying rod, wherein a through hole is arranged in the middle of the container vessel, and the lower end of the through hole corresponds to a biological material loading part of the carrying rod; the upper end of the container is provided with a liquid outlet groove, the interior of the container is provided with a liquid outlet cavity, and the liquid outlet cavity is respectively communicated with the liquid outlet groove and the through hole. The invention is convenient for processing cells before freezing, is convenient for quickly and accurately placing the cells into the through holes, and is also convenient for the cells to accurately fall onto the carrying rod in the later period, so that the freezing liquid wrapping the biological material has small and stable volume, the operation is more convenient and quicker, the cells do not need to be moved, and the operation steps are saved; the liquid outlet groove is connected with the through hole through the liquid outlet cavity, so that various vitrification solutions can be sucked away conveniently, the used vitrification solutions can be treated more easily and conveniently, the efficiency is higher, and the stability, timeliness and safety of operation are integrally guaranteed.)

1. A cell freezing assisting device comprises a carrying rod (1), and is characterized in that: the biological material loading device is characterized by further comprising a container vessel (2) attached to the upper end of the carrying rod (1), wherein a through hole (3) is formed in the middle of the container vessel (2), and the lower end of the through hole (3) corresponds to the biological material loading part of the carrying rod (1); the upper end of the container vessel (2) is provided with a liquid outlet groove (4), a liquid outlet cavity (5) is arranged inside the container vessel (2), and the liquid outlet groove (4) and the through hole (3) are respectively communicated with the liquid outlet cavity (5).

2. A cell freezing aid as claimed in claim 1 wherein: one end of the liquid discharge cavity (5) is communicated with the lower part of the liquid discharge groove (4), the other end of the liquid discharge cavity (5) surrounds the periphery of the through hole (3), a plurality of liquid discharge channels (6) are uniformly distributed between the through hole (3) and the liquid discharge cavity (5), and each liquid discharge channel (6) is communicated with the through hole (3) and the liquid discharge cavity (5).

3. A cell freezing aid as claimed in claim 2, wherein: each liquid drainage channel (6) is tangent to the through hole (3).

4. A cell freezing aid as claimed in claim 2 or 3 wherein: the container dish (2) comprises a transparent observation layer (201) positioned at the upper end of the liquid discharge cavity (5), and the through hole (3) and the liquid outlet groove (4) penetrate through the transparent observation layer (201).

5. A cell freezing aid according to claim 4 wherein: the upper part of the through hole (3) corresponding to the transparent observation layer (201) is of a funnel-shaped structure with the opening caliber gradually reduced from top to bottom.

6. A cell freezing aid according to claim 4 wherein: the container vessel (2) also comprises a solution treatment layer (202) positioned at the lower end of the transparent observation layer (201) and a release layer (203) positioned at the lower end of the solution treatment layer (202), and the upper end of the carrying rod (1) is attached to the lower end of the release layer (203); the liquid discharge cavity (5) and all the liquid discharge channels (6) are grooves arranged at the upper end of the solution treatment layer (202); the release layer (203) is made of an elastic material.

7. A cell freezing aid according to claim 6 wherein: the middle part of the through hole (3) corresponding to the solution processing layer (202) is of a funnel-shaped structure with the opening caliber gradually reduced from top to bottom.

8. A cell freezing aid according to claim 6 or 7 wherein: the container dish (2) is provided with a plurality of air holes (7), and the lower end of each air hole (7) is correspondingly arranged on the carrying rod (1).

9. A cell freezing aid as claimed in claim 8 wherein: the distance between each air hole (7) and the central axis of the through hole (3) is gradually reduced from top to bottom.

10. A cell freezing aid as claimed in claim 8 wherein: the lower end of the separation layer (203) is provided with an annular gas groove (8) positioned outside the through hole (3), and the lower end of each gas hole (7) is communicated with the annular gas groove (8).

Technical Field

The invention belongs to the technical field of biological material freezing, and particularly relates to a cell freezing auxiliary device.

Background

The art of vitrification cryopreservation of human and animal embryos is currently a relatively mature art, where "freezing" is liquid to solid cooling, which may include crystallization, and "vitrification" is liquid to solid cooling, but not crystallization. The vitrifying cryopreservation of human and animal embryos consists in the steps of collection and retrieval of the oocytes, their in vitro fertilization and subsequent storage of such fertilized eggs and the embryos and/or the resulting later blastocysts in an ultra-low temperature environment after treatment in a cryoprotective solution. The factors that influence the freezing process include osmotic shock and toxic shock of the freezing solution, knowledge and skill of the operator, and performance of the operating tools, such as a tool for freezing and loading cells, also known as a loading rod. The good carrier rod can ensure that the cells can obtain higher freezing and thawing speed and operability.

Carrier rods are available on the market as hollow tubes, or loop/hook type devices, such as Cryylogic, sold under the trade name fibreplug or Cryyloop, as defined in WO 00/21365. More typically other tools "Cryotop" as disclosed in international application WO 02/085110. Cryotop is an elastic strip of plastic joined to a grip. Wherein the cells were placed on a strip and then directly plunged into liquid nitrogen. At present, Cryotop is a mainstream technology with high survival rate and relative easy operability.

There are also many brand cryoprotectants on the market, with the principle being largely the same and different, in order to inhibit the formation of ice crystals within the cells and to minimize cell damage during the freezing process. These cryoprotectants are classified as osmotic and non-osmotic solutions. Examples of permeability are Ethylene Glycol (EG), dimethyl sulfoxide (DMSO), and glycerol. The osmotic cryoprotectant is a small molecule that readily penetrates the biofilm, forming hydrogen bonds with water molecules of the biomaterial, preventing its ice crystallization. Impermeable cryoprotectants, such as disaccharides, trehalose, and sucrose, function by extracting free water from within the biological material and dehydrating the intracellular space. The resulting dehydration allows it to complement the osmotic cryoprotectant to increase the intracellular relative concentration of the cryoprotectant, and to prevent the formation of ice crystals within the cell. However, the toxicity of these high concentrations of cryoprotectants can be considerable and the cells need to be rapidly plunged into liquid nitrogen after cryoprotectant pretreatment to achieve freezing. The very fast cooling rate minimizes the negative impact of the cryoprotectant on the biological material and also minimizes ice crystal formation due to imparting the desired vitrification.

The vitrification process involves exposing the biological material to at least three vitrification solutions. The vitrification solution is typically added to successive wells of a multi-well culture dish, wherein the culture dish and solution are warmed to a predetermined temperature, which is determined according to the requirements of the biological material under investigation.

In a typical protocol, the biological material is physically transferred to a first solution (e.g., ES equilibrator) in a first well, and then washed by physically moving the biological material or cells through the solution of interest using a cell pipetting device. The washing process is repeated in a second solution (e.g., VS vitrified cryo fluid), third and fourth wells in a second well for a predetermined period of time until the biological material or cells are considered ready for cryopreservation. The biological material is then physically aspirated with a predetermined amount of vitrification solution using a pipette or other manipulation device. The droplets containing the biological material or cells to be vitrified are then pipetted onto a vitrification device, such as a slide bar Cryotop. The vitrification device with the attached droplets and biological material is then physically transferred and directly plunged into liquid nitrogen. Once the biological material and carrier fluid are vitrified, the vitrification device is inserted into a pre-cooled protective sleeve or other storage device for subsequent transfer to liquid nitrogen or liquid nitrogen vapor for long-term cold storage.

Disclosure of Invention

In order to solve the above problems of the prior art, the present invention aims to provide a cell freezing assisting device.

The technical scheme adopted by the invention is as follows:

a cell freezing auxiliary device comprises a carrying rod and a container vessel attached to the upper end of the carrying rod, wherein a through hole is formed in the middle of the container vessel, and the lower end of the through hole corresponds to a biological material loading part of the carrying rod; the upper end of the container is provided with a liquid outlet groove, the interior of the container is provided with a liquid outlet cavity, and the liquid outlet cavity is respectively communicated with the liquid outlet groove and the through hole.

Preferably, one end of the liquid discharge cavity is communicated with the lower part of the liquid outlet groove, the other end of the liquid discharge cavity surrounds the periphery of the through hole, a plurality of liquid discharge channels are uniformly distributed between the through hole and the liquid discharge cavity, and each liquid discharge channel is communicated with the through hole and the liquid discharge cavity.

It is further preferred that each drainage channel is tangential to the through-going hole.

Still further preferably, the vessel includes a transparent observation layer at the upper end of the liquid discharge cavity, and the through hole and the liquid discharge groove both penetrate through the transparent observation layer.

It is further preferable that the upper portion of the through hole corresponding to the transparent observation layer is a funnel-shaped structure with an opening diameter gradually decreasing from top to bottom.

More preferably, the container further comprises a solution treatment layer positioned at the lower end of the transparent observation layer and a release layer positioned at the lower end of the solution treatment layer, and the upper end of the carrying rod is attached to the lower end of the release layer; the liquid discharge cavity and all the liquid discharge channels are grooves arranged at the upper end of the solution treatment layer; the release layer is made of an elastic material.

It is further preferable that the solution treatment layer has a funnel-shaped structure in which the opening diameter of the through hole gradually decreases from top to bottom.

Still further preferably, the container is provided with a plurality of air holes, and the lower end of each air hole is correspondingly arranged on the carrying rod.

It is further preferred that the distance between each air hole and the central axis of the through-hole is gradually smaller from top to bottom.

It is further preferable that the lower end of the release layer is provided with an annular gas groove located outside the through hole, and the lower end of each gas hole is communicated with the annular gas groove.

The invention has the beneficial effects that:

the invention is convenient for processing cells before freezing, increases the container, and arranges the through hole on the container, thus the size of the through hole can be properly enlarged, the cells can be conveniently and rapidly and accurately placed in the through hole, and the cells can accurately fall on the carrying rod in the later period, so the operation is more convenient and rapid, the cells do not need to be moved, and the operation steps are saved; the liquid outlet groove is connected with the through hole through the liquid outlet cavity, so that various vitrification solutions can be sucked away conveniently, the used vitrification solutions can be treated more easily and conveniently, the efficiency is higher, and the stability, timeliness and safety of operation are integrally guaranteed.

Drawings

FIG. 1 is a schematic view of an exploded structure of the vessel of the present invention;

FIG. 2 is a schematic view of the three-dimensional structure of the vessel of the present invention viewed from below;

FIG. 3 is a cross-sectional view of the vessel of the present invention;

fig. 4 is a front view of the present invention.

In the figure: 1-a carrier bar; 2-container vessel; 201-transparent viewing layer; 202-solution treatment layer; 203-a release layer; 3-a through hole; 4-a liquid outlet groove; 5-a liquid discharge cavity; 6-a liquid drainage channel; 7-pores; 8-annular gas groove.

Detailed Description

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the present invention will be briefly described below with reference to the accompanying drawings and the embodiments or the description in the prior art, it is obvious that the following description of the structure of the drawings is only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

The technical solution provided by the present invention will be described in detail by way of embodiments with reference to the accompanying drawings. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto.

In some instances, some embodiments are not described or not in detail, as they are conventional or customary in the art.

Furthermore, the technical features described herein, or the steps of all methods or processes disclosed, may be combined in any suitable manner in one or more embodiments, in addition to the mutually exclusive features and/or steps. It will be readily appreciated by those of skill in the art that the order of the steps or operations of the methods associated with the embodiments provided herein may be varied. Any order in the drawings and examples is for illustrative purposes only and does not imply that a certain order is required unless explicitly stated to be required.

The numbering of the components as such, e.g., "first", "second", etc., is used herein only to distinguish the objects as described, and does not have any sequential or technical meaning. The terms "connected" and "coupled" when used in this application, encompass both direct and indirect connections (and couplings) where appropriate and where not necessary contradictory.

It is an object of the present invention to provide a vessel that can be used for micromanipulation and storage of biological materials, covered on a CryoTop rod, to form a micro-vessel on the CryoTop rod, and after the cell solution has been processed, to remove the vessel to eliminate the cell migration steps, including but not limited to culture and cryopreservation of these materials. It is another object of the present invention to facilitate the use of automation to control the number of washing protocols and reduce handling of embryos, thereby achieving full automation. And the biomaterial of the present invention may be a fertilized egg or an oocyte.