CN209898090U - Sperm cryopreservation carrier and culture injection dish set - Google Patents

Abstract

The utility model provides a carrier and culture injection ware suit are preserved in sperm refrigeration, include: the cryopreservation carrier comprises a bearing part and a freezing chamber, wherein the bearing part is used for bearing the frozen micro-droplets, and at least one end edge of the bearing part is provided with a raised clamping part; a culture injection dish comprises a concave dish body, wherein a groove is formed in the inner surface of the bottom of the dish body, a bearing part of a cryopreservation carrier can be embedded into the groove, the groove is limited, the cryopreservation carrier is arranged, and a clamping part protrudes out of the inner surface of the bottom of the dish body. The utility model has the advantages of, the cryopreservation carrier just in time can block into the recess of culture injection ware, and the cryopreservation carrier can not take place to remove in the ware, and is stable, reliable, and the sight is clear, easily snatchs the sperm.

Description

Sperm cryopreservation carrier and culture injection dish set

Technical Field

The utility model relates to an auxiliary reproduction field in the medical science especially relates to a carrier and culture injection ware suit are preserved in sperm refrigeration.

Background

Cryopreservation of semen and sperm (hereinafter collectively referred to as sperm) is an important component of in vitro fertilization-embryo transfer (IVF-ET, commonly known as tube infant) treatment programs, and is in great demand. In addition to some social factors such as frozen semen (for example, the male can not get semen from the hospital due to the reason on the day of taking ovum), the condition that the semen needs to be frozen also includes the backup of frozen semen of the patient with semen taking difficulty (preventing semen taking failure on the day of taking ovum), the repeated semen taking and freezing of the occult azoospermia patient (accumulated for the day of taking ovum for multiple times), the cryopreservation of the semen obtained by the operation of the azoospermia patient (avoiding that the semen needs to be twice operated when the semen is not enough on the day of taking ovum).

In tube infant treatment, there are generally two types of insemination after resuscitation of frozen sperm, In Vitro Fertilization (IVF) and Intracytoplasmic sperm injection (ICSI). IVF is mostly used for infertility due to female factors, while semen parameters of male factors are normal. ICSI therapy is mostly used for male infertility (e.g. male oligospermia, azoospermia, occult azoospermia, etc.) and patients with low fertilization rate of previous IVF or history of previous IVF fertilization failure. More specifically, ICSI insemination modalities are seen in two cases: 1. the male has less sperms and weak activity, and can not be fertilized by IVF, such as male patients with oligospermia, azoospermia, occult azoospermia and various reasons requiring sperm acquisition by operation, which are hereinafter referred to as the first kind of patients; 2. the routine examination of male semen is normal, but there is a history of past IVF fertilization failure or IVF low fertilization rate, hereinafter referred to as a second type of patient.

A freezing tube (hereinafter, abbreviated as visual) of 1.0-2.0mL is a sperm cryopreservation carrier which is widely used at present. With visual frozen semen, the recovered semen needs density gradient centrifugation, centrifugal washing and/or upstream method treatment to be used for insemination, the treatment method is complex, and each treatment step can cause partial sperm damage (quality) and loss (quantity). For patients who want to adopt IVF insemination mode after recovery, the problem of adopting visual frozen semen is not great, because the total amount of sperms of the patients is large, the motility is good, even if partial sperms are damaged and lost, the treated sperms generally have enough moving sperms for IVF insemination, but for patients who want to adopt ICSI insemination mode after recovery, the visual is not an ideal carrier. According to two situations, the number of sperms of the first type of patients is small, the motility is poor, after the recovery by visual freezing, the sperms are difficult to find enough moving sperms to carry out ICSI, so that the sperms are forced to be frozen when the sperms are taken by secondary operations in an ovum taking day or no sperms are available, and therefore, the visual can not meet the sperm freezing requirement of the patients; the second type of patients have a history of previous IVF fertilization failure or a history of previous IVF low fertilization rate, but the semen is normally checked conventionally, and only dozens of active sperms are actually needed after recovery (the number of eggs obtained in the period of the test tube baby is generally about 10), so that the Vial is used as a cryopreservation carrier, and is large and small in size, and if the carrier is suitable, multiple sperms can be obtained at one time for preservation, so that the value is higher. In addition, if the treatment process can be simplified (the treatment process is preferably used after thawing) after the sperms of the patients are recovered and the sperms can be used for ICSI without complex treatment, the work efficiency of an embryo laboratory can be greatly improved.

How to achieve cryopreservation of sperm of the first type of patient and the second type of patient by using the carrier is the focus of current research. For example, a currently adopted method is that a long sheet-shaped or groove-shaped freezing sheet made of glass or plastic is used for loading sperms, the resuscitation of the freezing sheet is completed in hot oil at 37 ℃, so that the freezing preservation of testicular sperms, epididymal sperms and tiny sperms in semen of a patient with occult azoospermia can be realized, but the sperm grabbing by the method needs to be completed in a special wet box with saturated humidity to ensure that freezing liquid droplets are not evaporated, the osmotic pressure of the freezing liquid is maintained to be stable, the operation process is time-consuming and extremely tedious, and the wide clinical application of the utility model is greatly limited. In order to solve the problem, another method adopted at present is to insert an ultrathin cryopreservation carrier into an oil-covered sperm culture dish to grab the sperm, and the method does not need a wet box, does not worry about the evaporation of microdroplets of refrigerating fluid, can meet the cryopreservation of trace sperm from various sources, and is not limited by the operation time. However, the carrier needs to be directly inserted into an oil-covered sperm culture dish for operation, so the carrier needs to be made very thin (about 0.01 mm), otherwise, the transparency is insufficient, and the sight line for grabbing sperm is influenced; more importantly, the needle lifting is affected after the thickness is increased, so that the micro-operation needle is difficult to move and even broken. Because the carrier is thin and operates by being inserted into oil, the carrier can easily move (flap or slide) during the process of capturing sperm before freezing and during the process of ICSI after resuscitation, which increases the difficulty of operation and results in poor operation experience. In addition, microinjection vessels used in ICSI of each embryo laboratory are different in size, and the popularization and the use of the carrier of the utility model in IVF clinic are limited to a certain extent.

SUMMERY OF THE UTILITY MODEL

The technical problem to be solved by the utility model is to provide a sperm cryopreservation carrier and a culture injection dish set.

In order to solve the problem, the utility model provides a carrier and culture injection ware suit are preserved in freezing of sperm, include: the cryopreservation carrier comprises a bearing part and a freezing chamber, wherein the bearing part is used for bearing the frozen micro-droplets, and at least one end edge of the bearing part is provided with a raised clamping part; a culture injection dish comprises a concave dish body, wherein a groove is formed in the inner surface of the bottom of the dish body, a bearing part of a cryopreservation carrier can be embedded into the groove, the groove is limited, the cryopreservation carrier is arranged, and a clamping part protrudes out of the inner surface of the bottom of the dish body.

In one embodiment, the cryopreservation carrier is a transparent cryopreservation carrier and the culture injection dish is a transparent culture injection dish.

In one embodiment, the cryopreservation carrier is an L-shaped structure, the carrying part is the bottom of the L-shaped structure, and the clamping part is the side of the L-shaped structure.

In one embodiment, the bearing part is horizontal.

In one embodiment, the thickness of the bearing part is equal to the depth of the groove.

In an embodiment, the shape of the bearing part matches the shape of the groove.

In one embodiment, an anti-slip structure is arranged on the outer side surface of the dish body.

In one embodiment, the anti-slip structure is a circle of insections surrounding the outer side surface of the dish body.

In an embodiment, the culture injection dish further comprises a dish cover, the dish cover covers the opening of the dish body, and the diameter of the dish cover is larger than that of the dish bottom of the dish body.

In one embodiment, the cryopreservation carrier has a length of 3-15 mm, a width of 2-10 mm and a thickness of 0.1-1.5 mm, the culture injection dish is of a circular structure, the diameter of the bottom of the dish body is 30-60 mm, the diameter of the cover of the dish body is 31-61 mm, the thickness of the bottom of the dish body is 0.5-1.5 mm, and the thickness of the groove is 0.1-1.5 mm.

The utility model has the advantages that the sperm freezing and storing carrier and the culture injection dish set of the utility model have no special requirements on the sperm freezing liquid, and each laboratory can completely follow the original use habit to select the sperm freezing liquid sold in the market (such as the sperm freezing liquid containing glycerin) or self-prepared (such as the sperm freezing liquid containing low-concentration sucrose); the utility model inserts the freezing preservation carrier into the oil-covered culture injection dish to grab the sperms, overcomes the defects of the prior art, and the freezing liquid droplet on the freezing preservation carrier can not evaporate, so that special moisturizing equipment is not needed, the operation is simple, and the popularization is easy; the cryopreservation carrier of the utility model can be just clamped into the groove of the culture injection dish, and the cryopreservation carrier can not move in the dish, is stable and reliable, has clear sight and is easy to grab sperms; the utility model can not only freeze single sperm and rare sperm of various sources, but also freeze sperm (second class patient) in normal semen of semen routine check, in short, the method of the utility model can meet the sperm freezing of all resuscitated ICSI insemination modes; the method has the advantages that whether single sperm and rare sperm are frozen or the sperm of a second type of patients are frozen, after recovery, the method can be directly used for ICSI insemination of the ovum without complex processing procedures such as gradient centrifugation, centrifugal washing, upstream and the like, the operation is simple and convenient, and the work efficiency of an embryo laboratory is greatly improved; the utility model also has the advantages of high recovery rate (95-100%), high recovery rate (more than 60%), and the like.

The utility model discloses a further advantage lies in, the thickness of freezing carrier of preserving is the same with the degree of depth of culture injection ware, and the thickness of freezing carrier of preserving and the thickness of culture injection ware groove part just in time equal to the thickness at the bottom of the ware promptly, so behind the recess is gone into to the carrier card, just in time complete, level at the bottom of the ware, the microinjection needle that snatchs the sperm need not lift the needle and can freely translate at the bottom of the ware, can not take place disconnected needle.

Drawings

FIG. 1 is a schematic diagram of an exploded cross-section of a sperm cryopreservation carrier and a culture injection dish set according to the present invention;

FIG. 2 is a schematic structural view of the cryopreservation vector;

FIG. 3 is a schematic top view of the vessel body of the culture injection dish;

fig. 4 is a schematic cross-sectional view of the cryopreservation carrier 1 embedded in the groove;

FIG. 5 is a schematic diagram of a single sperm cryograsping sperm-culture injection dish position;

FIG. 6 is a schematic of the sperm cell thawing-culture injection dish position.

Detailed Description

The following will explain in detail the embodiments of the sperm cryopreservation carrier and the culture injection dish set provided by the present invention with reference to the accompanying drawings.

The utility model provides a sperm cryopreservation carrier and a culture injection dish set. The sperm freezing not only comprises sperm freezing for performing sperm insemination after sperm routine inspection is normal but recovery, but also comprises various rare sperm freezing and single sperm freezing such as oligoteratism, azoospermia, occult azoospermia and the like. The sperm source comprises ejaculated semen and sperm from various operations such as testicular puncture, epididymal puncture and microscopic semen collection. In short, as long as the sperms of the resuscitated Intracytoplasmic sperm microinjection (ICSI) can be used, the cryopreservation carrier and the culture injection dish set of the utility model can be used for the sperm after freezing, preservation and resuscitating.

FIG. 1 is a schematic diagram of an exploded cross section of the sperm cryopreservation carrier and the culture injection dish of the present invention. Referring to fig. 1, the sperm cryopreservation carrier and the culture injection dish of the present invention comprises a cryopreservation carrier 1 and a culture injection dish 2.

Fig. 2 is a schematic view of the structure of the cryopreservation carrier 1. Referring to fig. 1 and 2, the cryopreservation carrier 1 includes a carrying portion 10. The carrying part 10 is used for carrying frozen micro-droplets. Specifically, the carrying part 10 can carry more than 1 (including 1) 0.1-10.0uL of droplets of the refrigerating fluid. The bearing part 10 is horizontal to avoid the frozen micro-droplets from rolling off. The shape of the carrying part 10 includes, but is not limited to, rectangle, square, circle or other shapes. At least one end edge of the bearing part 10 is provided with a convex clamping part 11. The clamping portion 11 protrudes upward along at least one end edge of the carrying portion 10. During the freezing and thawing operation, the holder 11 may be grasped with a suitable tool (e.g., forceps) to move the cryopreservation carrier 1. In this embodiment, the cryopreservation carrier 1 has an L-shaped configuration, the carrying part 10 is the bottom of the L-shaped configuration, and the holding part 11 is the side of the L-shaped configuration. In other embodiments of the present invention, the shape of the cryopreservation carrier 1 can be other shapes as long as the object of the present invention can be achieved.

Fig. 3 is a schematic plan view of the dish body of the culture injection dish 2. Referring to fig. 1 and 3, the culture injection dish 2 includes a concave dish body 20. A groove 21 is arranged on the inner surface of the bottom of the dish body 20. Specifically, the groove 21 is provided on the inner surface of the bottom of the capsule 20, and does not penetrate the bottom of the capsule 20.

The carrier part 10 of the cryopreservation carrier 1 can be inserted into the groove 21. Fig. 4 is a schematic cross-sectional view of the cryopreservation carrier 1 embedded in the groove 21. Referring to fig. 4, the bearing portion 10 is embedded in the groove 21, and the groove 21 limits the cryopreservation carrier 1. That is, the bearing part 10 is inserted into the groove 21, and the groove 21 fixes the bearing part 10 to prevent the cryopreservation carrier 1 from moving when the sperm cryopreservation carrier and the culture dish set are used. The shape of the carrying part 10 matches the shape of the recess 21 to facilitate the engagement of the two, for example, if the carrying part 10 is rectangular, the recess 21 is also rectangular.

In this embodiment, the thickness of the bearing portion 10 is equal to the depth of the groove 21, that is, the bearing portion 10 is embedded in the groove 21, the upper surface of the bearing portion 10 is flush with the inner surface of the bottom of the dish body 20, and the thickness of the bearing portion 10 plus the thickness of the dish body 20 at the bottom of the groove 21 is exactly equal to the thickness of the bottom of the dish body 20, so that after the bearing portion 10 is clamped into the groove 21, the bottom of the dish body 20 is exactly complete and horizontal, and the microinjection needle for capturing sperms can freely translate at the bottom of the dish body 20 without lifting the needle, and needle breakage does not occur.

The clamping part 11 protrudes out of the inner surface of the bottom of the dish body 20 so as to be convenient for clamping. In this embodiment, the holding portion 11 may be disposed at an end of the carrying portion 10 facing the sidewall of the capsule 20 to prevent the holding portion 11 from affecting the movement of the micro-injection needle.

Further, the groove 21 is provided on one side of the bottom of the capsule 20 to provide a larger space for subsequent operations. Wherein the cryopreservation carrier 1 is a transparent cryopreservation carrier, the culture injection dish 2 is a transparent culture injection dish, and the cryopreservation carrier 1 and the culture injection dish 2 are made of transparent glass or plastic, for example. The spindle observation system is matched for use, the glass dish bottom is favorable for observing the spindle of the ovum during ICSI, and the success rate of ICSI treatment is improved.

Further, with continuing reference to fig. 1 and 3, an anti-slip structure 22 is disposed on an outer side surface of the vessel body 20 to facilitate the vessel body 20 to be more firmly taken out. Specifically, in the present embodiment, the anti-slip structure 22 is a circle of insections surrounding the outer side surface of the dish body, and in other embodiments, the anti-slip structure may be another structure capable of playing an anti-slip role, such as a plurality of microstructures arranged at intervals. The culture injection dish 2 further comprises a dish cover 23, the dish cover 23 covers the opening of the dish body 20 to cover the dish body 20, preferably, the diameter of the dish cover 23 is slightly larger than that of the dish bottom of the dish body 20, namely, the dish cover 23 can cover the dish body 20 and can be taken down easily.

Further, the length of the cryopreservation carrier 1 is 3-15 mm, the width is 2-10 mm, the thickness is 0.1-1.5 mm, the culture injection dish 2 is of a circular structure, the diameter of the bottom of the dish body 20 is 30-60 mm, the diameter of the dish cover 23 is 31-61 mm, the thickness of the bottom of the dish body 20 is 0.5-1.5 mm, and the thickness of the groove 21 is 0.1-1.5 mm.

The utility model discloses in, the sperm culture dish before the freezing both can be regarded as to the culture injection dish 2, also can regard as the microinjection dish after the recovery, and sperm culture dish and microinjection dish are one kind of dishes dual-purpose, and it is preserved carrier 1 with freezing and is constituteed a suit.

The utility model has the advantages of, (1) the utility model discloses carrier and culture injection dish suit are preserved in freezing of sperm does not have special requirement to the sperm refrigerating fluid, and each laboratory can follow the use custom completely and select to sell (if contain the sperm refrigerating fluid of glycerine) or from joining in marriage (if contain the sperm refrigerating fluid of low concentration sucrose). (2) The utility model discloses snatch the sperm in inserting the culture injection dish of lid oil with the cryopreservation carrier, overcome prior art's shortcoming, the refrigerating fluid droplet on the cryopreservation carrier can not evaporate, so need not to use special moisturizing equipment, easy operation, easy popularization. (3) The cryopreservation carrier can be just in time blocked into the groove of the culture injection dish, and the cryopreservation carrier can not move in the dish, is stable and reliable, has clear sight and is easy to grab sperms. (4) The thickness of the cryopreservation carrier is the same as the depth of the culture injection dish, namely the thickness of the cryopreservation carrier plus the thickness of the groove part of the culture injection dish is just equal to the thickness of the dish bottom, so that the carrier is clamped into the groove, the dish bottom is just complete and horizontal, and the micro-injection needle for grabbing sperms can freely translate at the dish bottom without lifting the needle, and the needle breakage can not occur. (5) The utility model discloses except can freezing single sperm and the rare sperm of various sources, can also freeze sperm (the second class patient) in the normal semen of semen routine inspection, in short, utilize the method can satisfy all after resumeing the sperm refrigeration of imitating going ICSI insemination mode. (6) No matter single sperm and rare sperm are frozen or sperm of a second type of patients are frozen, after recovery, the method can be directly used for ICSI insemination of the ovum without complex processing procedures of gradient centrifugation, centrifugal washing, upstream and the like, is simple and convenient to operate, and greatly improves the working efficiency of an embryo laboratory. (7) The utility model also has the advantages of high recovery rate (95-100%), high recovery rate (more than 60%), and the like.

The present invention will be further described with reference to the following specific examples. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. The materials, reagents, consumables, and the like used in the following examples are commercially available or may be prepared by oneself or by self-preparation, unless otherwise specified. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example 1

The utility model discloses the cryopreservation carrier and the culture injection dish suit include cryopreservation carrier and culture injection dish.

The cryopreservation carrier, also called a freezing piece, is a rectangular sheet structure, and can carry more than 1 (including 1) 0.1-10.0uL freezing liquid microdroplets in principle, and can be placed in a 0.5-2.0mL commercial freezing tube (visual), and any size meeting the two conditions can be adopted. The culture injection dish is of a circular structure, and in principle, the size of the culture injection dish can meet the requirement of an embryo laboratory for performing micro-insemination on ova, and meanwhile, a groove can be arranged at a proper position and the cryopreservation carrier can be placed in the culture injection dish. During freezing and recovery operation, the clamping part of the cryopreservation carrier is clamped by using a proper tool (such as tweezers), so that the cryopreservation carrier can be conveniently clamped and placed into or taken out of the groove of the culture injection dish.

Preferably, the long cryopreservation carrier has a length of 3-15 mm, a width of 2-10 mm and a thickness of 0.1-1.5 mm.

As a preferred scheme, the culture injection dish is of a circular structure, the diameter of the bottom of the dish is 30-60 mm, the diameter of the dish cover is 31-61 mm (the diameter of the dish cover is slightly larger than the diameter of the bottom of the dish, namely the dish cover can cover the dish body and can be taken down easily), the thickness of the bottom of the dish is 0.5-1.5 mm, the thickness of the groove part of the bottom of the dish is 0.1-1.5 mm, and the height of the dish body and the height of the dish cover are suitable.

In a more preferred embodiment, the cryopreservation carrier 1 is a rectangle having a length of 15mm, a width of 9mm, and a thickness of 0.5 mm. The diameter of the bottom of the dish body 20 of the culture injection dish 2 is 50mm, the diameter of the dish cover 23 is 51mm, the thickness of the bottom of the dish is 1mm, the length of the groove 21 is 15.2mm, the width of the groove 21 is 9.2mm, and the thickness of the groove 21 is 0.5 mm. The height of the dish body 20 of the culture injection dish 2 is 8mm, and the height of the dish cover 23 is 3 mm.

EXAMPLE 2 Monosperm freezing, thawing and intracytoplasmic sperm injection

The single sperm freezing refers to that the moving sperms are grabbed one by one under an inverted microscope by a microscopic injection needle (ICSI injection needle) and then transferred into refrigerating fluid microdroplets of a cryopreservation carrier for freezing, more than 1 (including 1) sperms can be frozen in each refrigerating fluid microdroplet, and the number of the frozen sperms can be counted. Monosperm freezing is not to say that only 1 sperm is frozen on one cryopreservation vehicle, in fact, typically 1-10 sperm are frozen on one cryopreservation vehicle, depending on the total number of motile sperm in the sample and the number of eggs available to the female. When the total number of motile sperm in a sperm sample is less than 200, it is recommended to grab the motile sperm frozen by a single sperm freezing process. Patients with non-obstructive azoospermia and occult azoospermia often need to freeze very minute amounts of sperm using a single sperm freezing method.

During freezing, 3-5 long drops are made of 10% SSS-mHTF above the groove of the culture injection dish and covered with mineral oil, and the sperm sample obtained by operation is properly processed and placed in the long drops for later use. Sperm refrigerating fluid of Origio is diluted 1:1, 0.5-1.0 mu L of refrigerating fluid droplets are prepared on a cryopreservation carrier, and the freezing fluid droplets are inserted into a groove of a culture injection dish. Using a microinjection needle ICSI injection needle, 1-10 motile sperm cells were grabbed into a cryo-droplet. Clamping the clamping part of the cryopreservation carrier by using a plastic forceps, taking out the cryopreservation carrier from the groove, removing the mineral oil on the carrier to the greatest extent by using oil absorption paper, cooling the liquid nitrogen steam for 2-5 minutes until the frozen liquid droplets on the carrier are frozen, adding liquid nitrogen, placing the carrier in 1.0-2.0mL of a commercially available freezing pipe (visual) in the liquid nitrogen, and storing in a liquid nitrogen tank for a long time.

FIG. 5 is a schematic diagram of a sperm-culture injection dish position for sperm freezing and grasping, with reference to FIG. 5, with 3-5 long droplets above groove 21 being 10% SSS-mHTF for storing sperm samples; 1 drop of the long-liquid dropping method is 7% -10% PVP, and the PVP is used for moistening and washing the micro-injection needle. After the micro-injection needle is rinsed in PVP, the movable sperms in the long liquid drops can be directly grabbed to the freezing liquid micro-drops on the carrier for storage and preparation for freezing.

When unfreezing, on the day of taking eggs from female, 50mL of mineral oil at 39 ℃ is prepared in advance for standby, and meanwhile, a culture injection dish is prepared for micro insemination. Before micro-insemination, a cryovial (visual) is taken out from a liquid nitrogen tank, a cryopreservation carrier is taken out by using a plastic forceps and is rapidly inserted into 50mL of mineral oil at 39 ℃, after a cryovial droplet on the cryopreservation carrier is dissolved (about 5 seconds), the cryopreservation carrier is inserted into a groove of an injection culture dish, and sperm is searched for in ICSI under an inverted microscope of 200 times.

FIG. 6 is a schematic diagram of the position of a single sperm thawing-culture injection dish, please refer to FIG. 6, the drop in the center of the grid is 7% -10% PVP for sperm braking; the surrounding 8 droplets were 10% SSS-mHTF for egg storage and injection; the lower droplet was 10% SSS-mHTF, and the resuscitated sperm on the cryopreservation vehicle was rinsed in this droplet, transferred into PVP and braked for ovum ICSI.

Example 3

Rare sperm freezing, thawing and intracytoplasmic sperm injection

The total number of the motile sperms obtained by the operation of most azoospermia patients is more than 200, for example, 1-2 motile sperms can be seen under every 200 times of high-power visual field, or 1 motile sperm can be seen under 2-3 200 times of high-power visual field, the sample size is about 50-100 muL or more, and the sperm can be frozen by adopting the method of freezing the rare sperm.

The culture injection dish is not needed to be used when the frozen. Properly processing a sperm sample obtained by an operation, fixing the volume to 50-100 mu L, adding sperm refrigerating fluid of an equivalent Origio company, uniformly mixing, adding 5-10 mu L of sperm-refrigerating fluid suspension on each cryopreservation carrier, placing the suspension in a 1.0-2.0mL (visual) commercial refrigerating tube, fixing the refrigerating tube on an aluminum frame, fumigating for 5 minutes by liquid nitrogen steam, adding liquid nitrogen after the suspension is frozen, and transferring the suspension into a liquid nitrogen tank for long-term storage.

The rare sperm thawing and intracytoplasmic sperm insemination was the same as the sperm thawing and intracytoplasmic sperm insemination in example 2, and need not be described in detail. The position schematic diagram of the rare sperm thawing-culture injection dish is the same as that of the single sperm thawing-culture injection dish.

Example 4

Sperm freezing, thawing and intracytoplasmic sperm injection in patients of the second type

In the treatment of test tube infants, about 5% -10% of patients adopting IVF insemination mode have fertilization failure or low fertilization rate (less than or equal to 30%), namely, men have normal semen routine examination, but people with past history of IVF fertilization failure or history of IVF low fertilization rate are referred to as second type of patients for short. At present, the sperm of the patients are frozen by adopting a traditional 1.0-2.0mL freezing tube (visual) to store the sperm, the recovered sperm can be used for insemination after being processed by density gradient centrifugation, centrifugal washing, and/or an upstream method, and the like, and the processing mode is complex. In fact, the ovum taking of the period of the patients needs to adopt an ICSI mode for insemination, and only about dozens of movable sperms are really used after recovery. By adopting the freezing preservation carrier of the patent to load the sperms and then freezing, one semen sample can be preserved in multiple parts, and the sperms can be directly used for ICSI without complex treatment after recovery, thereby greatly improving the work efficiency of the embryo laboratory.

The culture injection dish is not needed to be used when the frozen. Mixing semen sample 50 μ L with equal amount of sperm freezing solution from Origio company, adding semen-freezing solution suspension 5 μ L per freezing storage carrier, placing in commercial freezing tube (via) 1.0-2.0mL, fixing the freezing tube on aluminum frame, fumigating with liquid nitrogen steam for 5 min, freezing, adding liquid nitrogen, and transferring into liquid nitrogen tank for long-term storage. 6 carriers are frozen for each semen sample, and the semen sample can be used for 6 times of ovum taking. The sperm thawing and the micro insemination of the patients have the same single sperm and rare sperm without repeated description.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of improvements and decorations can be made without departing from the principle of the present invention, and these improvements and decorations should also be regarded as the protection scope of the present invention.

Claims (10)

1. The utility model provides a sperm cryopreservation carrier and culture injection dish suit which characterized in that includes:

the cryopreservation carrier comprises a bearing part and a freezing chamber, wherein the bearing part is used for bearing the frozen micro-droplets, and at least one end edge of the bearing part is provided with a raised clamping part;

a culture injection dish comprises a concave dish body, wherein a groove is formed in the inner surface of the bottom of the dish body, a bearing part of a cryopreservation carrier can be embedded into the groove, the groove is limited, the cryopreservation carrier is arranged, and a clamping part protrudes out of the inner surface of the bottom of the dish body.

2. The sperm cryopreservation carrier and culture injection dish set as claimed in claim 1, wherein the cryopreservation carrier is a transparent cryopreservation carrier and the culture injection dish is a transparent culture injection dish.

3. The sperm cryopreservation carrier and culture vessel set as in claim 1, wherein the cryopreservation carrier is of an L-shaped structure, the bearing portion is the bottom of the L-shaped structure, and the clamping portion is the side of the L-shaped structure.

4. The sperm cryopreservation carrier and culture vessel set as in claim 1, wherein the bearing part is horizontal.

5. The sperm cryopreservation carrier and culture dish set as in claim 1, wherein the thickness of the bearing part is equal to the depth of the groove.

6. The sperm cryopreservation carrier and culture vessel set as in claim 1, wherein the shape of the bearing portion matches the shape of the groove.

7. The sperm cryopreservation carrier and culture vessel set as in claim 1, wherein an anti-slip structure is arranged on the outer side surface of the vessel body.

8. The sperm cryopreservation carrier and culture vessel set as in claim 7, wherein the anti-slip structure is a ring of insections surrounding the outer side of the vessel body.

9. The sperm cryopreservation carrier and culture injection dish set as claimed in claim 1, wherein the culture injection dish further comprises a dish cover, the dish cover covers the opening of the dish body, and the diameter of the dish cover is larger than that of the dish bottom of the dish body.

10. The sperm cryopreservation carrier and culture injection dish set as claimed in claim 9, wherein the length of the cryopreservation carrier is 3-15 mm, the width is 2-10 mm, the thickness is 0.1-1.5 mm, the culture injection dish is of a circular structure, the diameter of the dish bottom of the dish body is 30-60 mm, the diameter of the dish cover is 31-61 mm, the thickness of the dish bottom of the dish body is 0.5-1.5 mm, and the thickness of the groove is 0.1-1.5 mm.

Images