CN102742568A - Apparatus for cryopreservation of biological sample - Google Patents
Apparatus for cryopreservation of biological sample Download PDFInfo
- Publication number
- CN102742568A CN102742568A CN2011100960426A CN201110096042A CN102742568A CN 102742568 A CN102742568 A CN 102742568A CN 2011100960426 A CN2011100960426 A CN 2011100960426A CN 201110096042 A CN201110096042 A CN 201110096042A CN 102742568 A CN102742568 A CN 102742568A
- Authority
- CN
- China
- Prior art keywords
- pipe
- cryovial
- mouth
- biological sample
- freezing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Agricultural Chemicals And Associated Chemicals (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention relates to an apparatus for cryopreservation of biological sample. The apparatus comprises a pipe (3); two ends of the pipe (3) are respectively provided with a matched nozzle (2); the pipe (3) is in a serpentine bending or spiral bending, or is embedded into a disk through a bend sleeve and is connected through a pipeline. The invention has the advantage that on the premise of ensuring freezing / melting uniformity, scale of slow / fast sample cryopreservation is enlarged, so as to avoid repeated operations in an existing technology.
Description
Technical field
The present invention relates to freezing field, particularly, the present invention relates to a kind of device that the biological sample cryogenic freezing is preserved that is used for.
Background technology
Freezing preservation is living beings article effective means of preserving steady in a long-term, also is the business-like keys of biological products.Under the cryogenic freezing state, biological metabolism almost stops life and keeps getting off with relative static conditions, and theoretically, under freezing conditions, the holding time of organism is unlimited.
At present freezing preserved two kinds of methods: slow freezing and snap frozen are the vitrifying method.The vitrifying method has been applied to egg mother cell and embryo's etc. freezing preservation; And slow freezing is little because of the relation of refrigerating effect and manual operation condition etc., and receives increasing concern.For zooblast, cell motility rate after it is freezing and frozen cooling speed are closely-related with melting heating rate.
In refrigerating process, because freezing rate is improper, cause ice crystal generation in the cell, finally cause the mechanical wounding of cell membrane; And in temperature-rise period,, original ice-nucleus is grown up because heating rate is slower, permeates cell membranes causes irreversible injury.For most of biological products, ice crystal dangerous temperature district is at 0～-60 ℃ of low-temperature region at present, and control refrigerating process and temperature-rise period speed are to improve the key of freezing preservation success rate.
At present, biological cryogenic freezing is preserved the refrigerated container that generally uses and is comprised cryovial, plastics or glass straw.Cryovial is the cylinder test tube of a band blind nut, and its material is generally polypropylene, polyethylene or polyphenylene oxide, and diameter 5～20mm does not wait, and volume is at 0.5ml～10ml.When freezing a large amount of sample, because the restriction of freezing equipment and thawing equipment etc., conditional during the height of cryovial; Can not infinitely extend, so the general cryovial packing form that adopts low capacity, the amount of labour is big like this; The sample treatment time is long, possibly cause the infringement to biological products.So in order to increase refrigerating capacity one time; Then need increase the cryovial diameter; Yet the increasing of diameter; With matter of utmost importance be cooling and the inhomogeneity of heating rate of the institute of Biological Products experience of diverse location in the cryovial, this will directly cause the heterogeneity of freezing preservation sample quality, the repeatability of properties of product is poor.So the freezing preservation pipe that present slow freezing uses is many in 10ml.What snap frozen was used is the elongated pipe-straw of diameter at 0.1～6.5mm plastics or glass, length 10～20cm, and useful load is not waited by 0.1～5ml.Its both ends open adopts steel ball or polyethylene powders to fill sealing; Usually heat up 4200 ℃ of warm speed/more than the min of 2500 ℃ of straw maximum instantaneous rate of temperature fall/more than the min, maximum instantaneous.Although straw can be realized the freezing of homogeneous and be rapidly heated speed that the process sealing is poor, is prone to cause pollute; And one time useful load is little, is difficult for scale and amplifies.
Therefore; The cylinder test tube of present biological sample cryoprotection; Its length can not infinitely prolong, but can cause the cooling of institute of Biological Products experience and the inhomogeneity of heating rate after the enlarged, so in this neck field; All do not have for a long time to solve the problem that refrigerating plant is difficult for amplification, limited the further promotion and application that change method.Therefore, a kind of freezing device of large-scale biological sample of realizing of design has great importance to the development of this area.
Summary of the invention
The objective of the invention is in order to overcome the problems referred to above; A kind of device that is used for the freezing preservation of biological sample is provided; Guaranteeing under freezing/intensification homogeneous condition; Can be used for the snap frozen of various content volume samples and the novel cryovial that slow freezing is preserved, realized the amplification of the freezing scale of sample.
To achieve these goals, the device that is used for the freezing preservation of biological sample of the present invention comprises a bend pipe 3, and the two ends of this bend pipe 3 are respectively equipped with the mouth of pipe 2 that matches.
Described bend pipe 3 is serpentine bend, spiral bending or be nested into disk through the different pipe of some diameters after be communicated with through pipeline.
Distance is 1～20mm between the described bend pipe 3 adjacent channel tube walls.
The described mouth of pipe 2 is the screwed mouth of pipe, and communicates nozzle diameter 0.2～2mm with the pipe pipe shaft.
Described device also comprises the mouthpiece cover 4 that is complementary with the mouth of pipe 2, and mouthpiece cover 4 inboards are threaded, and its internal diameter is identical with the external diameter of the mouth of pipe 2, can be tightened against on the mouth of pipe 2.
Described device also comprises a pipeline connector, and the two ends of this pipeline connector are connected with syringe or pipettor coupling respectively at the mouth of pipe 2, realizes that liquid injects or taking-up.
Described cryovial body top is two threaded mouths of pipe 2; The pipe shaft of described cryovial 1 bottom is made up of the pipeline winding or the stack of a series of bendings; Between its adjacent channel apart between the adjacent tube wall of pipe shaft of cryovial body bottom the distance 1～20mm; Internal diameter of tube body 0.1～20mm, body wall thickness 0.05～2mm.
Described mouthpiece cover 4 is made by low temperature resistant biocompatible materials with cryovial 1, polypropylene for example, polyethylene, polyvinyl chloride, polyphenylene oxide, glass; Described cryovial can be put within the cryopreservation equipment, and described cryovial can be put into freezing, also can independently place.
The biological sample freezing storage device that is used for of the present invention connects the cryovial mouth of pipe and syringe or pipettor through pipeline connector, realizes that liquid injects or taking-up; Concrete method for using may further comprise the steps: (1) sample injects: back out the cryovial lid; Connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector; Liquid in syringe or the pipettor is injected the cryovial body; Note during this time not producing bubble, amount of liquid should not surpass body volume 80%, removes pipeline connector and syringe or pipettor after the injection; The cryovial lid is tightened against the mouth of pipe, then can be directly or drop into liquid nitrogen after the precooling or directly put into other freezing equipments and carry out freezing preservation; Also can it be packed into after freezing according to the freezing preservation of above-mentioned approach; (2) sample takes out: after cryovial is placed 37 ℃ of quick-thawings; Back out the cryovial lid; Connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector, in the sucking-off cryovial body liquid or with air in syringe or the pipettor inject the cryovial body with liquid in the cryovial body from another mouth of pipe release.
The present invention and common heat transmission equipment have obvious district separated, and its operating temperature is low, and operation requires simple; But range of application is extremely extensive, and the present invention overcomes inertial thinking, changes conventional low-temperature biological refrigerating plant; This device is divided into straight tube and bend pipe two parts, and wherein bent tube section is taked to spiral or serpentine bend, make this device at a slow speed/the freezing preservation scale of quick sample obtain amplify; Have favorable uniformity simultaneously, thoroughly overcome the drawback of conventional apparatus, obtain good effect; Simultaneously because air section or bubble are brought in the expansion of the existence of bend pipe and capacity possibly cause the liquid injection time into; For this reason, employing one pipeline connector of the present invention makes it be connected with the syringe coupling with the mouth of pipe; Form airtight system at this section when liquid injects, thereby well overcome this problem.Therefore, thereby the present invention is through enlarging biological freezing scale to cryovial design cleverly, and the development in this field is had great importance.
The snap frozen of various content volume samples and novel cryovial and the method for using thereof that slow freezing is preserved of being used for provided by the invention.Compared with prior art, have the following advantages and beneficial effect: guaranteeing to realize under the freezing/homogeneous condition of melting at a slow speed/amplification of the freezing preservation scale of quick sample, avoiding the repetitive operation under the prior art condition.
Description of drawings
Fig. 1 a～c is the structural representation of the mouthpiece cover of apparatus of the present invention;
Fig. 2 is the structural representation of the cryovial of apparatus of the present invention;
Fig. 3 a～c is the different mouth of pipe type of a cryovial sketch map;
Fig. 4 a～b is apparatus of the present invention cryovial bottom schematic view;
Fig. 5 directly immerses liquid nitrogen frozen and 37 degrees centigrade of water-bath thaw point curves and test point sketch map for cryovial;
Fig. 6 is-80 degrees centigrade of ultra low temperature freezer cryogenic temperature curve maps.
The accompanying drawing sign
1, cryovial 2, the mouth of pipe 3, bend pipe
4, mouthpiece cover
Embodiment
Below in conjunction with accompanying drawing device of the present invention is further explained.
As shown in Figure 2; The snap frozen of various content volume samples and the novel refrigerating plant that slow freezing is preserved of being used for provided by the invention; Said device comprises: a bend pipe 3; The two ends of this bend pipe 3 are respectively equipped with the mouth of pipe 2 that matches, shown in Fig. 3 a～c the mouth of pipe 2 can for flanged, have sealing ring or have the mouth of pipe of flange or sealing ring simultaneously, described bend pipe 3 is serpentine bend, spiral bending or be nested into disk through bend pipe after be communicated with through pipeline.
The described mouth of pipe 2 is the screwed mouth of pipe, and communicates nozzle diameter 0.2～2mm with the pipe pipe shaft.
Described device also comprises the mouthpiece cover 4 that is complementary with the mouth of pipe 2, and shown in Fig. 1 a～c, mouthpiece cover 4 can be for the inboard be threaded, and the outside is threaded or the sealing ring mouthpiece cover is arranged very much, and its internal diameter is identical with the external diameter of the mouth of pipe 2, can be tightened against on the mouth of pipe 2.Described device also comprises a pipeline connector, and the two ends of this pipeline connector are connected with syringe or pipettor coupling respectively at the mouth of pipe 2, realizes that liquid injects or taking-up.
Described cryovial body top is two threaded mouths of pipe 2; The pipe shaft of described cryovial 1 bottom is made up of the pipeline winding or the stack of a series of bendings; Between its adjacent channel apart between the adjacent tube wall of pipe shaft of cryovial body bottom the distance 1～20mm; Internal diameter of tube body 0.1～20mm, body wall thickness 0.05～2mm.
Shown in Fig. 4 a～b, device of the present invention can be designed as base or does not have two kinds of base, can select as required.
The present invention realizes through following technical scheme: connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector, carry out liquid injection or taking-up.
The biological sample freezing storage device that is used for of the present invention connects the cryovial mouth of pipe and syringe or pipettor through pipeline connector, realizes that liquid injects or taking-up; Concrete method for using may further comprise the steps: (1) sample injects: back out the cryovial lid; Connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector; Liquid in syringe or the pipettor is injected the cryovial body; Note during this time not producing bubble, amount of liquid should not surpass body volume 80%, removes pipeline connector and syringe or pipettor after the injection; The cryovial lid is tightened against the mouth of pipe, then can be directly or drop into liquid nitrogen after the precooling or directly put into other freezing equipments and carry out freezing preservation; Also can it be packed into after freezing according to the freezing preservation of above-mentioned approach; (2) sample takes out: after cryovial is placed 37 ℃ of quick-thawings; Back out the cryovial lid; Connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector, in the sucking-off cryovial body liquid or with air in syringe or the pipettor inject the cryovial body with liquid in the cryovial body from another mouth of pipe release.
Its operation principle:
Sample injects: back out the cryovial lid; Connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector, liquid in syringe or the pipettor injected the cryovial body, during attention do not produce bubble; Amount of liquid should not surpass body volume 80%; Remove pipeline connector and syringe or pipettor after the injection, the cryovial lid is tightened against the mouth of pipe, shelve back, back input liquid nitrogen and carry out freezing preservation.
Sample takes out: after cryovial is placed 37 ℃ of quick-thawings; Back out the cryovial lid; Connect the cryovial mouth of pipe and syringe or pipettor through pipeline connector, in the sucking-off cryovial body liquid or with air in syringe or the pipettor inject the cryovial body with liquid in the cryovial body from another mouth of pipe release.
As shown in Figure 5, for cryovial directly immerses liquid nitrogen frozen and 37 degrees centigrade of water-baths when thawing, adopt thermocouple to measure the temperature curve of gained to the cryovial different parts.The cryovial internal diameter 6mm that adopts, wall thickness 1mm, useful load 25ml.Can know by figure, under the snap frozen condition, for the cryovial different parts, its temperature all at 2 to 3 seconds by reducing to more than 0 ℃ below-90 ℃ (0～-60 ℃ for low temperature deathtrap); Its maximum instantaneous rate of temperature fall all reach 2700 ℃/more than the min; The maximum instantaneous heating rate also all reach 4800 ℃/more than the min, suitable with 1～5ml straw, be higher than existing cryovial; Its useful load then is more than 1～5ml straw several times, shows good snap frozen performance.
As shown in Figure 6, be-80 degrees centigrade of ultra low temperature freezer cryogenic temperature curves.The cryovial internal diameter 6mm that adopts, wall thickness 1mm, useful load 25ml, 1.5ml cryovial internal diameter 1mm, wall thickness 0.2mm, useful load 1.5ml.Can know by figure, under the slow freezing condition, in 0～-60 ℃ of temperature range; For the cryovial different parts, 9 ℃/min of its average rate of temperature fall is slightly larger than 8 ℃/min of 1.5ml cryovial; And its variations in temperature basic synchronization, useful load is more than the cryovial several times, has embodied good homogeneity.
Therefore; Pass through improvement among the present invention to device; Overcome the medium-term and long-term problem that is difficult to overcome that exists of conventional refrigerating plant, and the data from Fig. 5 and Fig. 6 can find out that with analyzing this device has good snap frozen and homogeneous performance; And capacity greatly increases, and therefore has good effect.
It should be noted last that above embodiment is only unrestricted in order to technical scheme of the present invention to be described.Although the present invention is specified with reference to embodiment; Those of ordinary skill in the art is to be understood that; Technical scheme of the present invention is made amendment or is equal to replacement, do not break away from the spirit and the scope of technical scheme of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (8)
1. a device that is used for the freezing preservation of biological sample is characterized in that, this device comprises a bend pipe (3), and the two ends of this bend pipe (3) are respectively equipped with the mouth of pipe (2) that matches.
2. the device of the freezing preservation of biological sample according to claim 1 is characterized in that, described bend pipe (3) is serpentine bend, spiral bending or be nested into disk through some pipes after be communicated with through pipeline.
3. the device of the freezing preservation of biological sample according to claim 2 is characterized in that, distance is 1～20mm between described bend pipe (3) the adjacent channel tube wall.
4. the device of the freezing preservation of biological sample according to claim 1 is characterized in that, the described mouth of pipe (2) is the screwed mouth of pipe.
5. the device of the freezing preservation of biological sample according to claim 1 is characterized in that, described device also comprises the mouthpiece cover (4) that is complementary with the mouth of pipe (2).
6. the device of the freezing preservation of biological sample according to claim 1; It is characterized in that; Described device also comprises a pipeline connector, and the two ends of this pipeline connector are connected with syringe or pipettor coupling respectively at the mouth of pipe (2), realizes that liquid injects or taking-up.
7. the device of the freezing preservation of biological sample according to claim 1 is characterized in that, described bend pipe (3) internal diameter 0.1～20mm, body wall thickness 0.05～2mm.
8. the device of the freezing preservation of biological sample according to claim 1; It is characterized in that; Described cryovial (1), the mouth of pipe (2) and mouthpiece cover (4) adopt low temperature resistant biocompatible materials to process, and comprise polypropylene, polyethylene, polyvinyl chloride, polyphenylene oxide and glass.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100960426A CN102742568A (en) | 2011-04-18 | 2011-04-18 | Apparatus for cryopreservation of biological sample |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100960426A CN102742568A (en) | 2011-04-18 | 2011-04-18 | Apparatus for cryopreservation of biological sample |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102742568A true CN102742568A (en) | 2012-10-24 |
Family
ID=47023346
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011100960426A Pending CN102742568A (en) | 2011-04-18 | 2011-04-18 | Apparatus for cryopreservation of biological sample |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102742568A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104642298A (en) * | 2013-11-21 | 2015-05-27 | 浙江星博生物科技有限公司 | Miniature loading system for cryopreservation of biological materials |
| CN106342789A (en) * | 2016-10-13 | 2017-01-25 | 深圳韦拓生物科技有限公司 | Cell vitrification freezing treatment equipment and method |
| CN106455540A (en) * | 2014-03-31 | 2017-02-22 | 通用电气健康护理生物科学股份公司 | Product and method for cell preservation |
| CN107650412A (en) * | 2017-10-27 | 2018-02-02 | 合肥燕庄食用油有限责任公司 | A kind of squeezer for cold pressing sesame oil |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101031635A (en) * | 2004-08-06 | 2007-09-05 | 社团法人家畜改良事业团 | Biosample freezing apparatus and method of freezing |
| WO2007108536A1 (en) * | 2006-03-23 | 2007-09-27 | Axis International Co., Ltd. | Body fluid leakage preventive for dead body and dead body treating equipment using the same |
| CN101386814A (en) * | 2008-10-21 | 2009-03-18 | 北京大学第三医院 | Biological material vitrified frozen vector and preparation method thereof |
| WO2010056755A2 (en) * | 2008-11-11 | 2010-05-20 | Craig H Randall | Microfluidic embryo and gamete culture systems |
| CN201789875U (en) * | 2010-09-01 | 2011-04-13 | 湖南光琇高新生命科技有限公司 | Large-scale stem cell vitrifying and freezing tube |
-
2011
- 2011-04-18 CN CN2011100960426A patent/CN102742568A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101031635A (en) * | 2004-08-06 | 2007-09-05 | 社团法人家畜改良事业团 | Biosample freezing apparatus and method of freezing |
| WO2007108536A1 (en) * | 2006-03-23 | 2007-09-27 | Axis International Co., Ltd. | Body fluid leakage preventive for dead body and dead body treating equipment using the same |
| CN101386814A (en) * | 2008-10-21 | 2009-03-18 | 北京大学第三医院 | Biological material vitrified frozen vector and preparation method thereof |
| WO2010056755A2 (en) * | 2008-11-11 | 2010-05-20 | Craig H Randall | Microfluidic embryo and gamete culture systems |
| CN201789875U (en) * | 2010-09-01 | 2011-04-13 | 湖南光琇高新生命科技有限公司 | Large-scale stem cell vitrifying and freezing tube |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104642298A (en) * | 2013-11-21 | 2015-05-27 | 浙江星博生物科技有限公司 | Miniature loading system for cryopreservation of biological materials |
| CN106455540A (en) * | 2014-03-31 | 2017-02-22 | 通用电气健康护理生物科学股份公司 | Product and method for cell preservation |
| CN106455540B (en) * | 2014-03-31 | 2021-03-19 | 思拓凡瑞典有限公司 | Products and methods for cell preservation |
| US10959423B2 (en) | 2014-03-31 | 2021-03-30 | Cytiva Sweden Ab | Product and method for cell preservation |
| CN106342789A (en) * | 2016-10-13 | 2017-01-25 | 深圳韦拓生物科技有限公司 | Cell vitrification freezing treatment equipment and method |
| CN107650412A (en) * | 2017-10-27 | 2018-02-02 | 合肥燕庄食用油有限责任公司 | A kind of squeezer for cold pressing sesame oil |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104677000B (en) | Liquid nitrogen cooling cryogenic device and implementation method for same | |
| CN102742568A (en) | Apparatus for cryopreservation of biological sample | |
| CN112105863B (en) | Method and device for filling dry type dewar tank | |
| CN101694338A (en) | Ice bag capable of applying in preserving biological product repeatedly and method for preparing the ice bag | |
| CN111457768B (en) | Be applied to frozen low temperature phase transition heat exchanger of cell vitrification | |
| CN104012521B (en) | A kind of contactless sessile drop method refrigerating plant and freezing method | |
| CN206504516U (en) | High-precision cooling system | |
| CN201136864Y (en) | Programmed cell freezing instrument with quick temperature reduction and temperature recovery functions | |
| CN106386786A (en) | Freezing preservation container for low-temperature-sensitive biological sample | |
| CN208925062U (en) | A kind of fast cooling equipment | |
| CN201126087Y (en) | Heat preserving heat insulating slab | |
| CN206310815U (en) | Double source cold storage box | |
| CN202773904U (en) | Biological material vitrification freezing carrier | |
| CN204568405U (en) | A kind of Cord blood test tube stand transfer box | |
| CN206207829U (en) | Quick-frozen ice chest | |
| CN101514859B (en) | Multifunctional refrigerating and quick freezing integrated storeroom | |
| CN200989866Y (en) | Cryogenic freeze dryer for biological medical experiment | |
| CN2682849Y (en) | Small-sized low-temperature creature freezing instrument | |
| CN204022794U (en) | A kind of beer fermenting device and beer preparation system | |
| CN207095078U (en) | A kind of CO2 gas refrigerations device | |
| CN206906075U (en) | A kind of low temperature samples pipe | |
| CN203186902U (en) | Refrigeration preservation system | |
| CN201637217U (en) | Ice-temperature and quick-frozen integral warehouse | |
| CN202535988U (en) | Cooling control device for cell, tissue and organ low-temperature preservation | |
| CN204799313U (en) | A centrifuging tube keeps box for liquid nitrogen quick -freeze |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121024 |