CN105850986B - An efficient and safe human rare sperm DPV freezing system and freezing method - Google Patents

An efficient and safe human rare sperm DPV freezing system and freezing method Download PDF

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Publication number
CN105850986B
CN105850986B CN201610396509.1A CN201610396509A CN105850986B CN 105850986 B CN105850986 B CN 105850986B CN 201610396509 A CN201610396509 A CN 201610396509A CN 105850986 B CN105850986 B CN 105850986B
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droplet
liquid nitrogen
safety
safe
freezing
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CN105850986A (en
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陈威
孙健
张箴波
吴煜
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Shanghai First Peoples Hospital
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Shanghai First Peoples Hospital
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/10—Preservation of living parts
    • A01N1/14—Mechanical aspects of preservation; Apparatus or containers therefor
    • A01N1/142—Apparatus

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The present invention relates to a kind of people's rareness sperm DPV refrigeration systems of highly effective and safe, including safe droplet slide glass, freezing droplet and liquid nitrogen vapor tank, the safe droplet slide glass is made of biological safety sterile material, safe droplet slide glass is divided into several regions of the same area, and safe droplet slide glass is that unified material is integrally formed；The freezing droplet is made of the special cryoprotection drop of fixed capacity；Bottom is equipped with liquid nitrogen in the liquid nitrogen vapor tank, and liquid nitrogen vapor is full of in tank, and safe droplet slide glass is placed in freezing housing and is placed in liquid nitrogen vapor tank.The invention further relates to a kind of people's rareness sperm DPV freezing methods.Advantages of the present invention is embodied in: using the biological safety sterile material of optimum thickness, convenient for efficiently utilizing the frozen sperm on the safe droplet slide glass；Safe droplet slide glass is that unified material is integrally formed, and prevents other materials from polluting；With region division, more preferably it is convenient for long-term preservation, more saving in liquid nitrogen vapor prevents cross contamination.

Description

A kind of the people's rareness sperm DPV refrigeration system and freezing method of highly effective and safe
Technical field
The present invention relates to assisted reproduction fields, and in particular to a kind of people's rareness sperm DPV refrigeration system of highly effective and safe and Freezing method.
Background technique
The infertility of male factor has accounted for 40-50% at present, and there are also more serious development trends.It is more next The disease incidence of higher few azoospermia seriously affects the fecundity of male.Although we can be by testicular biopsy or micro- Cut obtain sperm, but at present lack highly effective and safe freezen protective rareness sperm, especially single sperm freezing carrier and Method.
Chinese patent (CN 104823966A) discloses a kind of micro sperm cryopreservation carrier and corresponding freezing reconciliation Jelly method, but its carrier medium is to lack safety and lack region division and can not effectively prevent intersecting dirt between sample Dye.
Summary of the invention
It is rare the purpose of the present invention is aiming at the shortcomings in the prior art, providing a kind of people of highly effective and safe no cross contamination Sperm (contains monosperm, as testicular biopsy sperm, epididymis puncture the micro- incision essence of sperm, Azoospermia patient sperm, testis Son etc.) DPV refrigeration system.
To achieve the above object, the invention discloses following technical solutions:
A kind of people's rareness sperm DPV refrigeration system of highly effective and safe, including safe droplet slide glass, freezing droplet and liquid nitrogen steam Vapour tank, the safe droplet slide glass are made of biological safety sterile material, and safe droplet slide glass is divided into several identical The region of area, safe droplet slide glass are that unified material is integrally formed；The freezing droplet is protected by the special freezing of fixed capacity It protects drop to constitute, freezing droplet is dripped when in use on safe droplet slide glass；Bottom is equipped with liquid nitrogen in the liquid nitrogen vapor tank, and And liquid nitrogen vapor is full of in tank, safe droplet slide glass is placed in freezing housing and is placed in liquid nitrogen vapor tank.
Further, the safe droplet slide glass is rectangular tab.
Further, the material of the safe droplet slide glass is thin using the biological safety polypropylene of ethylene oxide sterilizing Piece.
Further, the safe droplet slide glass with a thickness of 1.5 μm.
Further, the safe droplet slide glass is divided into 8 regions of the same area.
Further, the safe droplet slide glass is equipped with the handle of an identical material, handle and safe droplet slide glass one It is body formed.
Further, height of the liquid nitrogen liquid level that the liquid nitrogen vapor pot bottom is equipped with not higher than freezing housing.
Further, bracket is equipped in the liquid nitrogen vapor tank, safe droplet slide glass is placed in freezing housing and passes through Bracket is placed in liquid nitrogen vapor tank.
Another object of the present invention is to provide a kind of people's rareness sperm of highly effective and safe (containing monosperm, such as testicular biopsy essence Son, epididymis puncture sperm, Azoospermia patient sperm, micro- incision sperm of testis etc.) freezing and storing method, utilization is above-mentioned DPV refrigeration system, specifically comprise the following steps:
The micro sperm freezing housing of S1: drop has freezing droplet on safe droplet slide glass, is added in each freezing droplet corresponding Sperm after, by the safe droplet slide glass be put into in the matched freezing housing of its size with other sperm samples isolation open, so The freezing housing for being loaded with safe droplet slide glass liquid nitrogen is carried out afterwards to fumigate；
The micro sperm long-term preservation of S2: prepare the liquid nitrogen vapor tank that a bottom is equipped with liquid nitrogen, and keep liquid nitrogen vapor Liquid nitrogen vapor is full of in tank, the freezing housing for being loaded with safe droplet slide glass is rack-mount, it is placed on liquid nitrogen steaming for a long time It is saved in vapour.
The people's rareness sperm DPV refrigeration system and freezing method of a kind of highly effective and safe disclosed by the invention have with following Beneficial effect:
Safe droplet slide glass uses the biological safety sterile material of optimum thickness, convenient for efficiently being carried using the safe droplet The frozen sperm of on piece；
Safe droplet slide glass is that unified material is integrally formed, and can prevent other materials from polluting；
With region division, more preferably it is convenient for long-term preservation, and cross contamination between sample can be prevented.
Detailed description of the invention
Fig. 1 is the combination diagram of safe droplet slide glass and freezing housing；
Fig. 2 is the step schematic diagram (1) that sperm sample is packed into freezing droplet；
Fig. 3 is the step schematic diagram (2) that sperm sample is packed into freezing droplet
Fig. 4 is the step schematic diagram of micro sperm long-term preservation；
Fig. 5 is the step schematic diagram for taking out frozen sperm.
Description of symbols:
1. housing is freezed, 2. handles, 3. safe droplet slide glasses, 4. sperm samples, 5. freezing droplets, 6. brackets, 7. liquid nitrogen Vapor can, 8. liquid nitrogen.
Specific embodiment
Below with reference to embodiment and referring to attached drawing, the invention will be further described.
- Fig. 4 referring to Figure 1.
A kind of people's rareness sperm DPV refrigeration system of highly effective and safe, including safe droplet slide glass 1, freezing droplet 5 and liquid nitrogen Vapor can 7, the safe droplet slide glass 3 are made of biological safety sterile material, and safe droplet slide glass 3 is divided into several Region of the same area, convenient for efficiently utilizing the frozen sperm on the freezing carrier, safe droplet slide glass 3 is unified material one Molding, prevents other materials from polluting；The freezing droplet 5 is made of the special cryoprotection drop of fixed capacity, freezes droplet 5 It is dripped when in use on safe droplet slide glass 3；Bottom is equipped with liquid nitrogen 8 in the liquid nitrogen vapor tank 7, and liquid nitrogen is full of in tank Steam, safe droplet slide glass 3 are placed in freezing housing 1 and are placed in liquid nitrogen vapor tank 7.
As specific embodiment, the safe droplet slide glass 3 is Thin Rectangular slide glass.
As specific embodiment, the material of the safe droplet slide glass 3 is poly- using the biological safety of ethylene oxide sterilizing Propylene thin slice, the carrier are the biological safety sterile material of optimum thickness.
As specific embodiment, the safe droplet slide glass 3 with a thickness of 1.5 μm.
As specific embodiment, the safe droplet slide glass 3 is divided into 8 regions of the same area.
As specific embodiment, the safe droplet slide glass 3 is equipped with the handle 2 of an identical material, and handle 2 and safety are micro- Slide glass 3 is dripped to be integrally formed.
As specific embodiment, 8 liquid level of liquid nitrogen that 7 bottom of liquid nitrogen vapor tank is equipped with is not higher than freezing housing 1 Height.
See Fig. 4, as specific embodiment, is equipped with bracket 6 in the liquid nitrogen vapor tank 7, safe droplet slide glass 3 is placed on cold Freeze in housing 1 and is placed in liquid nitrogen vapor tank 7 by bracket 6.
A kind of people's rareness sperm cryopreservation method of highly effective and safe disclosed by the invention, using above-mentioned DPV refrigeration system, Include the following steps:
The micro sperm freezing housing of step 1: drop has freezing droplet 5 on safe droplet slide glass 3, adds in each freezing droplet 5 After entering corresponding sperm, by the safe droplet slide glass 3 be put into in the matched freezing housing 1 of its size with other sperm samples every It opens absolutely, the freezing housing 1 for being loaded with safe droplet slide glass 3 is then carried out liquid nitrogen and is fumigated.
The micro sperm long-term preservation of step 2: prepare the liquid nitrogen vapor tank 7 that a bottom is equipped with liquid nitrogen 8, and keep liquid nitrogen It is full of liquid nitrogen vapor in vapor can 7, on the frame 6 by the installation of freezing housing 1 for being loaded with safe droplet slide glass 3, places for a long time It is saved in liquid nitrogen vapor.
See Fig. 5, when taking out frozen sperm sample, takes out the freezing housing 1 for having safe droplet slide glass 3 first, will pacify Full droplet slide glass 3 is placed in sampling ware, and sampled instrument draws frozen sperm sample.
The present invention uses the biological safety sterile material of optimum thickness, convenient for efficiently utilizing on the safe droplet slide glass Frozen sperm；Safe droplet slide glass is that unified material is integrally formed, and can prevent other materials from polluting；With region division, more preferably Convenient for long-term preservation.
The above is only a preferred embodiment of the present invention, rather than its limitations；Although should be pointed out that referring to above-mentioned each Embodiment describes the invention in detail, those skilled in the art should understand that, it still can be to above-mentioned each Technical solution documented by embodiment is modified, or equivalent substitution of some or all of the technical features；And this A little modifications and replacement, do not make the essence of corresponding technical solution depart from the scope of the technical solutions of the embodiments of the present invention.

Claims (4)

1.一种高效安全的人稀少精子DPV冷冻系统，其特征在于，包括安全微滴载片、冷冻微滴和液氮蒸汽罐，所述安全微滴载片采用生物安全性灭菌材料制成，安全微滴载片被划分为8个相同面积的区域，安全微滴载片为统一材料一体成型；所述冷冻微滴由固定容量的特殊冷冻保护液滴构成，冷冻微滴在使用时滴于安全微滴载片上；所述液氮蒸汽罐内底部装有液氮，并且在罐内充满液氮蒸汽，安全微滴载片放置在冷冻外套内并放置在液氮蒸汽罐内；所述安全微滴载片为矩形薄片，安全微滴载片的材料采用环氧乙烷灭菌的生物安全性聚丙烯薄片，安全微滴载片的厚度为1.5μm；所述液氮蒸汽罐内设有支架，安全微滴载片放置在冷冻外套内并通过支架放置在液氮蒸汽罐内。1. an efficient and safe human rare sperm DPV freezing system, is characterized in that, comprises safety droplet slide, frozen droplet and liquid nitrogen steam tank, and described safety droplet slide adopts biosafety sterilization material to make , the safety droplet slide is divided into 8 areas of the same area, and the safety droplet slide is integrally formed with a uniform material; the frozen droplet is composed of special cryoprotective droplets with a fixed capacity, and the frozen droplet drops when used. on the safety droplet carrier; the liquid nitrogen vapor tank is equipped with liquid nitrogen at the bottom, and the tank is filled with liquid nitrogen vapor, and the safety droplet carrier is placed in the freezing jacket and placed in the liquid nitrogen vapor tank; the The safety droplet carrier is a rectangular sheet, the material of the safety droplet carrier is a biosafety polypropylene sheet sterilized by ethylene oxide, and the thickness of the safety droplet carrier is 1.5 μm; With a holder, the safety droplet slide is placed in a cryo jacket and through the holder into a liquid nitrogen vapor tank. 2.根据权利要求1所述的一种高效安全的人稀少精子DPV冷冻系统，其特征在于，所述安全微滴载片设有一个相同材料的把手，把手与安全微滴载片一体成型。2 . An efficient and safe DPV freezing system for rare human spermatozoa according to claim 1 , wherein the safety droplet slide is provided with a handle made of the same material, and the handle and the safety droplet slide are integrally formed. 3 . 3.根据权利要求1所述的一种高效安全的人稀少精子DPV冷冻系统，其特征在于，所述液氮蒸汽罐底部设有的液氮液面高度不高于冷冻外套的高度。3. A kind of efficient and safe human rare sperm DPV freezing system according to claim 1, is characterized in that, the height of the liquid nitrogen liquid level that the bottom of described liquid nitrogen steam tank is provided with is not higher than the height of freezing jacket. 4.一种高效安全的人稀少精子冷冻保存方法，其特征在于，利用如权利要求1-3所述的DPV冷冻系统，包括如下步骤：4. an efficient and safe cryopreservation method for rare human spermatozoa, is characterized in that, utilizes the DPV freezing system as claimed in claim 1-3, comprises the steps: S1微量精子冷冻外套：安全微滴载片上滴有冷冻微滴，各个冷冻微滴中加入相应的精子后，将该安全微滴载片放入与其大小匹配的冷冻外套中与其他精子样本隔绝开，然后把装载了安全微滴载片的冷冻外套进行液氮熏蒸；S1 Microsperm Freezing Jacket: Frozen droplets are placed on the safety droplet slide. After adding the corresponding sperm to each frozen droplet, the safety droplet slide is placed in a freezer jacket that matches its size and is isolated from other sperm samples. , and then liquid nitrogen fumigation was performed on the cryojacket loaded with the safety droplet slide; S2微量精子长期保存：准备一个底部装有液氮的液氮蒸汽罐，并且保持液氮蒸汽罐内充满液氮蒸汽，将装载了安全微滴载片的冷冻外套安装在支架上，长期放置在液氮蒸汽中进行保存。S2 Microsperm long-term storage: prepare a liquid nitrogen vapor tank with liquid nitrogen at the bottom, and keep the liquid nitrogen vapor tank filled with liquid nitrogen vapor, install the cryojacket loaded with the safety microdroplet slide on the holder, and place it on the shelf for a long time. stored in liquid nitrogen vapor.
CN201610396509.1A 2016-06-06 2016-06-06 An efficient and safe human rare sperm DPV freezing system and freezing method Expired - Fee Related CN105850986B (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20220053756A1 (en) * 2019-01-08 2022-02-24 Changzhou Maternal And Child Health Care Hospital Chip carrier for single sperm or extremely small amount of sperms
CN113040132B (en) * 2021-03-22 2022-03-25 浙江大学 Sperm cryopreservation method based on protective agent
CN113712025A (en) * 2021-09-14 2021-11-30 荆州市中心医院 Cryopreservation method of trace sperms and preparation method of sperms directly used for ICSI

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101671651A (en) * 2008-09-09 2010-03-17 上海交通大学医学院附属第九人民医院 Sperm freezing and unfreezing method and sperm freezing and unfreezing device
CN202514486U (en) * 2012-02-16 2012-11-07 上海安集协康生物技术有限公司 Container suitable for liquid nitrogen tank to store cells
CN104823966A (en) * 2015-05-19 2015-08-12 上海交通大学医学院附属第九人民医院 Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof
CN204888506U (en) * 2015-07-22 2015-12-23 柳州市妇幼保健院 Efficient biomaterial freezes and deposits carrier device

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101671651A (en) * 2008-09-09 2010-03-17 上海交通大学医学院附属第九人民医院 Sperm freezing and unfreezing method and sperm freezing and unfreezing device
CN202514486U (en) * 2012-02-16 2012-11-07 上海安集协康生物技术有限公司 Container suitable for liquid nitrogen tank to store cells
CN104823966A (en) * 2015-05-19 2015-08-12 上海交通大学医学院附属第九人民医院 Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof
CN204888506U (en) * 2015-07-22 2015-12-23 柳州市妇幼保健院 Efficient biomaterial freezes and deposits carrier device

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