CN106234351B - For the fluidization load of biological tissue or the device of taking-up cryoprotective agent - Google Patents
For the fluidization load of biological tissue or the device of taking-up cryoprotective agent Download PDFInfo
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- CN106234351B CN106234351B CN201610620317.4A CN201610620317A CN106234351B CN 106234351 B CN106234351 B CN 106234351B CN 201610620317 A CN201610620317 A CN 201610620317A CN 106234351 B CN106234351 B CN 106234351B
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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
- A01N1/02—Preservation of living parts
- A01N1/0236—Mechanical aspects
- A01N1/0242—Apparatuses, i.e. devices used in the process of preservation of living parts, such as pumps, refrigeration devices or any other devices featuring moving parts and/or temperature controlling components
- A01N1/0252—Temperature controlling refrigerating apparatus, i.e. devices used to actively control the temperature of a designated internal volume, e.g. refrigerators, freeze-drying apparatus or liquid nitrogen baths
- A01N1/0257—Stationary or portable vessels generating cryogenic temperatures
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Abstract
A kind of fluidization load for biological tissue or the device for taking out cryoprotective agent, it is characterized by: it includes solution loop module, solution supplying module, sample container, temperature control box and controller, the sample container is arranged in temperature control box, the sample container is equipped with inlet and liquid outlet, the solution loop module is connected with the inlet of sample container and liquid outlet so that solution circulation flow is through sample container, the solution supplying module is connected to supply solution to solution loop module with solution loop module, the controller respectively with solution supplying module, solution loop module and temperature control box electrical connection.The automation equipment can accelerate the rate that cryoprotective agent is loaded into and takes out biological tissue while reducing limit process temperature.
Description
Technical field
The present invention relates to cryo-conservation technical fields, and in particular to a kind of fluidization load or taking-up for biological tissue
The device of cryoprotective agent.
Background technique
Cryo-conservation, which refers to, is cooled to low temperature using special method for biological sample living, generally -196 DEG C simultaneously long-term
It saves；When needed, biological sample is heated to normal temperature by special method, still can get biological sample living.Low temperature
When saving biological sample, other than open-and-shut microorganism, virus and bacterium etc., it is however generally that addition cryoprotective agent is required
's.
Though cryoprotective agent can at low temperature shield to biological sample, work to this protection, it is necessary to
Enough cryoprotective agents are loaded into biological sample.However, cryoprotective agent is foreign substance for biological sample,
There is certain toxicity to it.The close phase of the factors such as this toxicity and type, concentration, treatment temperature and the time of cryoprotective agent
It closes, concentration is bigger, treatment temperature is higher, the processing time is longer, then toxic damages are more serious.Assuming that the low of biological sample need to be loaded into
The protectant concentration of temperature determines, it is evident that load temperature is lower, the load time is shorter, it is more advantageous to cryo-conservation.Unlike outstanding
Biological samples, the cryoprotective agent naturally osmotic biological tissues such as floating cell generally require longer time, cause to load low temperature guarantor
When protecting agent, biological tissue is more subject to toxic damages, this is also the difficult point place of current cryo-conservation biological tissue.Reduce load
Although temperature can reduce toxic damages, but simultaneously but also the rate of cryoprotective agent infiltration biological tissue slows down, this is meaned
Be loaded into same amount of cryoprotective agent and need the longer time, and the load time lengthens the wind that will increase by toxic damages
Danger.In addition, minimum load temperature is commonly considered as it and balances chill point temperature when the concentration of low temperature protectant solution determines,
It can freeze if being further continued for reducing, it means that under prior art conditions, reduce toxicity by reducing load temperature
Damage is limited.There is also the same problems when cryoprotective agent is taken out from biological tissue.
Therefore, if lowest limit treatment temperature can be made to be down to balance chill point temperature hereinafter, i.e. enhancing is subcooled, while can again
The rate for accelerating cryoprotective agent infiltration biological tissue, then can substantially reduce the toxic damages that biological tissue is subjected to, can make
The success rate of cryo-conservation biological tissue greatly improves, and the automation equipment still that this kind of function not can be achieved at present.
Summary of the invention
The technical problem to be solved by the present invention is low temperature can be accelerated while reducing limit process temperature by providing one kind
Protective agent is loaded into and takes out the automation equipment of the rate of biological tissue.
The technical solution of the invention is as follows: cryoprotective agent is taken out in a kind of fluidization load for biological tissue
Device, it is characterised in that: it includes solution loop module, solution supplying module, sample container, temperature control box and controller, described
Sample container is arranged in temperature control box, and the sample container is equipped with inlet and liquid outlet, the solution loop module and sample
The inlet of container is connected with liquid outlet so that solution circulation flow is recycled through sample container, the solution supplying module and solution
Module be connected with to solution loop module supply solution, the controller respectively with solution supplying module, solution loop module
It is electrically connected with temperature control box.
After adopting the above structure, the invention has the following advantages that
Low temperature protectant solution of the present invention becomes a kind of liquid of persistent loop flowing, this stream by solution loop module
Dynamic property, which makes low temperature protectant solution be reduced to balance chill point temperature or less in temperature, to be solidified, that is, it is existing supercooling occur
As so as to reduce limit process temperature；And this mobility can accelerate low temperature protectant solution infiltration biology simultaneously
The rate of tissue, so that the present apparatus can accelerate cryoprotective agent while reducing limit process temperature is loaded into and takes out life
The rate of object tissue compensates for the blank in this field；In addition to this, due to solution supplying module, solution loop module and temperature
Control case is controlled by controller, so that whole device realizes automation.
Preferably, further including gas supplying module, the sample container is additionally provided with air inlet and air outlet, the gas
Supplying module is connected with the air inlet of sample container, so as to produce into the gas in sample container to the solution in sample container
Raw disturbance is simultaneously discharged from gas outlet, and the gas supplying module is electrically connected with the controller.The setting passes through gas supplying module
The solution in sample container is set to generate disturbance, and because the mobility of solution can further decrease pole so that level of disruption enhances
Treatment temperature is limited, and accelerates the rate that cryoprotective agent is loaded into and takes out biological tissue.
Preferably, the sample container includes gas flow cavity and the sample cavity that is arranged in gas flow cavity, institute
It states inlet and liquid outlet is arranged on sample cavity, the air inlet and air outlet are arranged on gas flow cavity, the solution
Loop module is connected with the inlet of sample cavity and liquid outlet, the air inlet phase of the gas supplying module and gas flow cavity
It is connected to, the air hole of several permeable watertights is additionally provided on the sample cavity, so that the gas entered at air inlet can pass through sample
Product chamber is simultaneously discharged from gas outlet.The setting is independent by fluid passage and gas passage, and the solution in fluid passage will not
It enters in gas passage, but the gas in gas passage can enter in fluid passage the disturbance for enhancing solution, overall structure
Simply, reliable operation.
Be arranged in gas flow cavity preferably, the sample cavity extends transversely through, gas flow cavity is divided on
Half chamber and lower half chamber, the inlet and liquid outlet are separately positioned on the both ends of sample cavity, and the air inlet is arranged in gas
On the lower half chamber of body flow cavity, the gas outlet is arranged on the upper half chamber of gas flow cavity, and the air hole setting exists
On the side wall of sample cavity.The setting flows transversely through solution along sample cavity, and gas from bottom to top passes through sample cavity, so that disturbance
Effect is stronger, so as to largely reduce limit process temperature, and accelerates cryoprotective agent and is loaded into and takes out biological tissue
Rate.
Preferably, the first straight line L1 where the air inlet and air outlet is perpendicular to where inlet and liquid outlet
Second straight line L2.The setting is mutually perpendicular to fluid passage and gas passage, can utmostly enhance the disturbance of solution, so as to
Limit process temperature is utmostly reduced, and accelerates the rate that cryoprotective agent is loaded into and takes out biological tissue.
Preferably, several partitions are arranged in uniform intervals in transverse direction in the sample cavity.The setting can be by multiple lifes
Object tissue sample is separated without being overlapped, and the speed of biological tissue is loaded into and taken out so as to improve cryoprotective agent
Rate.
Preferably, the solution loop module include check valve, mix container, the first peristaltic pump, First Heat Exchanger and
Tapping valve, the check valve, mix container, the first peristaltic pump, First Heat Exchanger are sequentially connected along solution flow direction, described the
One heat exchanger is located in temperature control box, the inlet of the tapping valve inlet with the liquid outlet of sample container and check valve respectively
It is connected；The solution loop module is connected with the inlet of sample container and liquid outlet refers to the first of solution loop module
The liquid outlet of heat exchanger, the inlet of check valve are connected with the inlet of sample container, liquid outlet respectively；The controller with
The electrical connection of solution loop module refers to that controller is electrically connected with the first peristaltic pump.The setting can control low temperature protectant solution with
Certain flow velocity circulation flows through sample container, is conducive to improve rate and drop that cryoprotective agent is loaded into and takes out biological tissue
The limit process temperature of low low temperature protectant solution；Meanwhile solution can be recycled to save reagent, mix container and have both tune
It saves circulatory system volume, keep pressure and the function conducive to solution in the discharge circulatory system in the circulatory system.
Preferably, the solution supplying module is compacted including the first fluid reservoir, the second fluid reservoir, the second peristaltic pump, third
Dynamic pump, conductance electrode and platinum resistance temperature sensor, first fluid reservoir is connected with the inlet of the second peristaltic pump, described
Second fluid reservoir is connected with the inlet of third peristaltic pump, and the conductance electrode and platinum resistance temperature sensor are located at mixing and hold
In device；The solution supplying module is connected with solution loop module refers to the liquid outlet and third peristaltic pump of the second peristaltic pump
It is connected respectively with the inlet of the liquid outlet of check valve, mixing container after liquid outlet connection；The controller and solution supply
Module electrical connection refers to that controller is electric with the second peristaltic pump, third peristaltic pump, conductance electrode and platinum resistance temperature sensor respectively
Connection.This is provided with the low temperature protectant solution for being conducive to precisely supply required concentration to solution loop module.
Preferably, the gas supplying module includes gas generator, the second heat exchanger, gas generator is changed with second
Hot device is connected, and second heat exchanger is located in temperature control box；The gas supplying module is connected with the air inlet of sample container
The gas outlet for referring to the second heat exchanger is connected with the air inlet of sample container；The controller is electrically connected with gas supplying module
Refer to that controller is electrically connected with gas generator.The setting structure is simple and convenient to operate, and is more advantageous to implementation of the invention and is pushed away
Extensively.
Preferably, the gas in the gas supplying module is nitrogen.Nitrogen is cheap and using safe.
Detailed description of the invention:
Fig. 1 is the present invention for the fluidization load of biological tissue or the structural representation for the device for taking out cryoprotective agent
Figure；
Fig. 2 is the structural schematic diagram of sample container of the present invention；
In figure: the first fluid reservoir of 1-, the second fluid reservoir of 2-, the second peristaltic pump of 3-, 4- third peristaltic pump, 5- mix container,
6- conductance electrode, the first peristaltic pump of 7-, 8- First Heat Exchanger, 9- sample container, 9.1- sample cavity, 9.2- partition, 9.3- gas
Flow cavity, 9.4- inlet, 9.5- liquid outlet, 9.6- air inlet, the gas outlet 9.7-, 10- tapping valve, 11- check valve, 12- gas
Body generator, the second heat exchanger of 13-, 14- temperature control box, 15- controller, 16- platinum resistance temperature sensor, 17- upper half chamber,
18- lower half chamber, 19- air hole, L1- first straight line, L2- second straight line.
Specific embodiment
With reference to the accompanying drawing, and in conjunction with the embodiments the present invention is described further.
Embodiment:
As shown in Figure 1 and Figure 2, a kind of fluidization for biological tissue loads or takes out the device of cryoprotective agent, special
Sign is: it includes solution loop module, solution supplying module, sample container 9, temperature control box 14 and controller 15, the sample
Container 9 is arranged in temperature control box 14, and the sample container 9 is equipped with inlet 9.4 and liquid outlet 9.5, the solution loop module
It is connected with the inlet 9.4 of sample container 9 and liquid outlet 9.5 so that solution circulation flow is through sample container 9, the solution supply
Module is connected to supply solution to solution loop module with solution loop module, and the controller 15 supplies mould with solution respectively
Block, solution loop module and temperature control box 14 are electrically connected.
Low temperature protectant solution of the present invention becomes a kind of liquid of persistent loop flowing, this stream by solution loop module
Dynamic property, which makes low temperature protectant solution be reduced to balance chill point temperature or less in temperature, to be solidified, that is, it is existing supercooling occur
As so as to reduce limit process temperature；And this mobility can accelerate low temperature protectant solution infiltration biology simultaneously
The rate of tissue, so that the present apparatus can accelerate cryoprotective agent while reducing limit process temperature is loaded into and takes out life
The rate of object tissue compensates for the blank in this field；In addition to this, due to solution supplying module, solution loop module and temperature
It controls case 14 to be controlled by controller 15, so that whole device realizes automation.
Preferably, further including gas supplying module, the sample container 9 is additionally provided with air inlet 9.6 and gas outlet 9.7,
The gas supplying module is connected with the air inlet 9.6 of sample container 9, so that into the gas in sample container 9 to sample
Solution in container generates disturbance and is discharged from gas outlet 9.7, and the gas supplying module is electrically connected with controller 15.This sets
Set makes the solution in sample container generate disturbance by gas supplying module, and because of the mobility of solution, so that level of disruption
Enhancing, can further decrease limit process temperature, and accelerate the rate that cryoprotective agent is loaded into and takes out biological tissue.
Preferably, the sample container 9 includes gas flow cavity 9.3 and the sample being arranged in gas flow cavity 9.3
Product chamber 9.1, the inlet 9.4 and liquid outlet 9.5 are arranged on sample cavity 9.1, and the air inlet 9.6 and gas outlet 9.7 are set
It sets on gas flow cavity 9.3, the solution loop module is connected with the inlet 9.4 of sample cavity 9.1 and liquid outlet 9.5,
The gas supplying module is connected with the air inlet 9.6 of gas flow cavity 9.3, parches if being additionally provided on the sample cavity 9.1
The fluid-tight air hole 19 of gas, so that the gas entered at air inlet 9.6 can pass through sample cavity 9.1 and arrange from gas outlet 9.7
Out.The setting is independent by fluid passage and gas passage, and the solution in fluid passage will not enter in gas passage, but
Gas in gas passage can enter in fluid passage the disturbance for enhancing solution, and overall structure is simple, reliable operation.
It is arranged in gas flow cavity 9.3 preferably, the sample cavity 9.1 extends transversely through, by gas flow cavity 9.3
It is divided into upper half chamber 17 and lower half chamber 18, the inlet 9.4 and liquid outlet 9.5 are separately positioned on the two of sample cavity 9.1
End, the air inlet 9.6 are arranged on the lower half chamber 18 of gas flow cavity 9.3, and the setting of gas outlet 9.7 is flowed in gas
On the upper half chamber 17 of chamber 9.3, the air hole 19 is arranged on the side wall of sample cavity 9.1.The setting makes solution along sample cavity
9.1 flow transversely through, and gas from bottom to top passes through sample cavity 9.1, so that perturbation action is stronger, so as to largely reduce
Limit process temperature, and accelerate the rate that cryoprotective agent is loaded into and takes out biological tissue.
Preferably, the first straight line L1 where the air inlet 9.6 and gas outlet 9.7 perpendicular to inlet 9.4 and goes out
Second straight line L2 where liquid mouth 9.5.The setting is mutually perpendicular to fluid passage and gas passage, can utmostly enhance solution
Disturbance, so as to utmostly reduce limit process temperature, and accelerate cryoprotective agent be loaded into and take out biological tissue speed
Rate.
Preferably, several partitions 9.2 are arranged in uniform intervals in transverse direction in the sample cavity 9.1, to clearly show that
The structure of sample cavity 9.1 and partition 9.2, nitrogen flow chamber 9.3 only provide half in Fig. 2, and sample cavity 9.1 is also removed one
Point.The setting can be separated without being overlapped by multiple biological tissue samples, is loaded into so as to improve cryoprotective agent
With the rate for taking out biological tissue.
Preferably, the solution loop module includes check valve 11, mixes container 5, the heat exchange of the first peristaltic pump 7, first
Device 8 and tapping valve 10, the check valve 11, mix container 5, the first peristaltic pump 7, First Heat Exchanger 8 along solution flow direction according to
Sequence connection, the First Heat Exchanger 8 is located in temperature control box 14, the inlet of the tapping valve 10 respectively with sample container 9 out
Liquid mouth 9.5 is connected with the inlet of check valve 11；The inlet 9.4 of the solution loop module and sample container 9 and out liquid
Mouthfuls 9.5 inlets for being connected the liquid outlet, check valve 11 that refer to the First Heat Exchanger 8 of solution loop module hold with sample respectively
Inlet 9.4, the liquid outlet 9.5 of device 9 are connected；The controller 15 be electrically connected with solution loop module refer to controller 15 with
The electrical connection of first peristaltic pump 7.The setting can control low temperature protectant solution and flow through sample container 9 with certain flow velocity circulation,
Be conducive to improve cryoprotective agent loading and take out the rate of biological tissue and reduce the limit process of low temperature protectant solution
Temperature；Meanwhile solution can be recycled to save reagent, mix container 5 and have both adjusting circulatory system volume, keep cyclic system
Pressure and the function conducive to solution in the discharge circulatory system in uniting.
Preferably, the solution supplying module includes the first fluid reservoir 1, the second fluid reservoir 2, the second peristaltic pump 3, third
Peristaltic pump 4, conductance electrode 6 and platinum resistance temperature sensor 16, the inlet phase of first fluid reservoir 1 and the second peristaltic pump 3
Connection, second fluid reservoir 2 are connected with the inlet of third peristaltic pump 4, the conductance electrode 6 and platinum resistance temperature sensing
Device 16, which is located at, to be mixed in container 5；The solution supplying module is connected with solution loop module refers to the liquid out of the second peristaltic pump 3
Mouth is connected with the inlet of the liquid outlet of check valve 11, mixing container 5 respectively with after the connection of the liquid outlet of third peristaltic pump 4；
The controller 15 be electrically connected with solution supplying module refer to controller 15 respectively with the second peristaltic pump 3, third peristaltic pump 4, electricity
Conductive electrode 6 and platinum resistance temperature sensor 16 are electrically connected.This is provided with required dense conducive to precisely supplying to solution loop module
The low temperature protectant solution of degree.
Preferably, the gas supplying module includes gas generator 12, the second heat exchanger 13, gas generator 12 with
Second heat exchanger 13 is connected, and second heat exchanger 13 is located in temperature control box 14；The gas supplying module and sample container 9
Air inlet 9.6, which is connected, refers to that the gas outlet of the second heat exchanger 13 is connected with the air inlet 9.6 of sample container 9；The control
Device 15 is electrically connected with gas supplying module refers to that controller 15 is electrically connected with gas generator 12.The setting structure is simple, operates
It is convenient, it is more advantageous to implementation and popularization of the invention.
Preferably, the gas in the gas supplying module is nitrogen.Nitrogen is cheap and using safe.
Above-mentioned First Heat Exchanger 8 and the second heat exchanger 13 are bent by one section of longer pipeline to be made, and can also install fin additional
Deng the structure for being conducive to heat exchange, it is all can be realized fluid and flow through and carry out the structure of heat exchange be suitable for 8 He of First Heat Exchanger
Structure represented by second heat exchanger 13；Above-mentioned temperature control box 14 is the typical temperature control device of the prior art, can be mechanical refrigeration
Or liquid nitrogen refrigerating combination electric heater unit, temperature sensor are realized, understand that all components are by box table in figure for diagram brief introduction
Show；What the mixing container 5 was applied is the typical prior art, i.e. electromagnetic agitation, is understood for diagram brief introduction, stirrer and electromagnetism
Blender does not provide.Filled arrows indicate solution flow direction in figure, and hollow arrow indicates nitrogen flow direction.
The present invention is mainly made of four parts: solution concentration control, temperature control, solution flow control and nitrogen flow rate control
System.The effect of solution concentration control section be make circulation flow through sample container 9 low temperature protectant solution concentration according to setting
Change of program, including the first fluid reservoir 1, the second fluid reservoir 2, the second peristaltic pump 3, third peristaltic pump 4, conductance electrode 6, platinum electricity
Hinder temperature sensor 16 and controller 15.The effect of the temprature control unit is the cryoprotective agent for flowing through sample container 9
The temperature of solution and nitrogen is according to the change of program of setting, including temperature control box 14, First Heat Exchanger 8,13 and of the second heat exchanger
Controller 15.The effect of the solution flow rate control part is the stream for making to recycle the low temperature protectant solution for flowing through sample container 9
Speed is maintained at some definite value or the change of program according to setting, including the first peristaltic pump 7 and controller 15.The nitrogen flow rate
The effect of control section be flow through the nitrogen of sample container 9 flow velocity be maintained at some definite value or according to setting program become
Change, including nitrogen gas generator 12 and controller 15.
Before carrying out fluidization load to biological tissue samples or take out cryoprotective agent processing, first have to establish use
Relational model between the conductivity and concentration and temperature of low temperature protectant solution, and be input in controller 15.For diformazan
Sulfoxide/sodium chloride/water, glycerol/sodium chloride/water, ethylene glycol/sodium chloride/water, propylene glycol/sodium chloride/water, trehalose/chlorination
The concentration of this five kinds of low temperature protectant solutions of sodium/water, sodium chloride is definite value 0.9%wt, between conductivity and concentration and temperature
Relational model is as follows:
In formula, κ is conductivity, and unit: mS/cm, T are temperature, and unit: K, x are the mass fraction of cryoprotective agent, and R is
Mol gas constant, value are 8.314J/ (molK), κ0、Ea、C0、C1And C2For model coefficient, value is as shown in the table:
For the present invention for loading dimethyl sulfoxide into articular cartilage tissue, loading procedure is as follows:
Open the program that is built in controller 15, input dimethyl sulfoxide/sodium chloride/aqueous solution conductivity and concentration and
Relational model between temperature, and set concentration, temperature and change in flow program；By the sodium chloride solution and height of 0.9%wt
Concentration dimethyl sulfoxide/sodium chloride/aqueous solution is respectively charged into the first fluid reservoir 1 and the second fluid reservoir 2；One or more are big
Small suitable articular cartilage tissue is placed between 9.1 internal partition 9.2 of sample cavity；Open the first peristaltic pump 7, the second peristaltic pump 3,
Three peristaltic pumps 4, nitrogen gas generator 12 and temperature control box 14.
The concentration of dimethyl sulfoxide solution is by changing the second peristaltic pump 3 and third peristaltic pump in the solution loop module
4 revolving speed controls.The sodium chloride solution and dimethyl sulfoxide are mixed well in container 5 mixing, and conductance electrode 6 detects solution
Conductivity, platinum resistance temperature sensor 16 detects the temperature of solution, and the two transmits signals to controller 15 respectively and be converted into
Concentration values, and be compared with preset concentration, controller 15 outputs signals to the first peristaltic pump 7 and second according to processing result
Peristaltic pump 3 controls the revolving speed of the two.
For temperature according to the change of program of setting, the solution in solution loop module passes through the first heat exchange in the temperature control box 14
Enter in sample cavity 9.1 after device 8 is cooled, the nitrogen in the nitrogen supplying module is cooled by the second heat exchanger 13, stream
Through sample cavity 9.1 to generate disturbance to the solution in sample cavity 9.1, finally it is discharged from gas outlet 9.7.
The flow velocity of solution is controlled by changing the revolving speed of the first peristaltic pump 7 in the solution loop module, and described first
The revolving speed of peristaltic pump 7 keeps certain value or the change of program according to setting；The flow velocity of nitrogen is kept in the nitrogen supplying module
Certain value or change of program according to setting.
After loading procedure, the second peristaltic pump 3, third peristaltic pump 4, nitrogen gas generator 12 and temperature control box 14 are closed, is beaten
Tapping valve 10 is opened, the first peristaltic pump 7 and tapping valve 10 is closed after the solution in solution loop module empties, takes out articular cartilage
Tissue is to carry out subsequent processing；Distilled water is finally packed into the first fluid reservoir 1, the second peristaltic pump 3 is opened, by a certain amount of distillation
Water is conveyed into solution loop module, opens the first peristaltic pump 7 with distilled water flushing solution loop module and then opens tapping valve
10 empty flushing liquor, and device has been cleaned at this time, can be loaded next time.
The above are the loading procedures of present apparatus low temperature protectant solution, and it is similar with loading procedure to take out process, here not
It repeats again, the difference is that: the concentration of solution gradually rises in solution loop module in loading procedure, and temperature control box 14
Interior temperature is gradually reduced, and the concentration of solution gradually decreases in solution loop module during taking out, and temperature control
Temperature in case 14 is gradually increasing.
Claims (9)
1. a kind of fluidization for biological tissue loads or takes out the device of cryoprotective agent, it is characterised in that: it includes molten
Liquid loop module, solution supplying module, sample container (9), temperature control box (14) and controller (15), the sample container (9) set
It sets in temperature control box (14), the sample container (9) is equipped with inlet (9.4) and liquid outlet (9.5), the solution loop module
It is connected with the inlet (9.4) of sample container (9) and liquid outlet (9.5) so that solution circulation flow is through sample container (9), it is described
Solution supplying module be connected with solution loop module with to solution loop module supply solution, the controller (15) respectively with
Solution supplying module, solution loop module and temperature control box (14) electrical connection；It further include gas supplying module, the sample container
(9) air inlet (9.6) and gas outlet (9.7), the air inlet (9.6) of the gas supplying module and sample container (9) are additionally provided with
It is connected, so as to generate disturbance to the solution in sample container (9) into the gas in sample container (9) and from gas outlet
(9.7) it is discharged at, the gas supplying module is electrically connected with controller (15).
2. a kind of fluidization for biological tissue according to claim 1 loads or takes out the device of cryoprotective agent,
It is characterized by: the sample container (9) includes the sample of gas flow cavity (9.3) and setting in gas flow cavity (9.3)
Product chamber (9.1), the inlet (9.4) and liquid outlet (9.5) are arranged on sample cavity (9.1), the air inlet (9.6) and out
Port (9.7) is arranged on gas flow cavity (9.3), the inlet (9.4) of the solution loop module and sample cavity (9.1) and
Liquid outlet (9.5) is connected, and the gas supplying module is connected with the air inlet (9.6) of gas flow cavity (9.3), the sample
Product chamber (9.1) is equipped with the air hole (19) of permeable watertight, so that the gas entered at air inlet (9.6) can pass through sample cavity
(9.1) and at gas outlet (9.7) it is discharged.
3. a kind of fluidization for biological tissue according to claim 2 loads or takes out the device of cryoprotective agent,
It is characterized by: the sample cavity (9.1) extends transversely through setting in gas flow cavity (9.3), by gas flow cavity (9.3)
It is divided into upper half chamber (17) and lower half chamber (18), the inlet (9.4) and liquid outlet (9.5) are separately positioned on sample cavity
(9.1) both ends, the air inlet (9.6) are arranged on the lower half chamber (18) of gas flow cavity (9.3), the gas outlet
(9.7) it is arranged on the upper half chamber (17) of gas flow cavity (9.3), the air hole (19) is arranged in sample cavity (9.1)
On side wall.
4. a kind of fluidization for biological tissue according to claim 3 loads or takes out the device of cryoprotective agent,
It is characterized by: first straight line (L1) where the air inlet (9.6) and gas outlet (9.7) perpendicular to inlet (9.4) and
Second straight line (L2) where liquid outlet (9.5).
5. a kind of fluidization for biological tissue according to claim 3 loads or takes out the device of cryoprotective agent,
It is characterized by: several partitions (9.2) are arranged in uniform intervals in transverse direction in the sample cavity (9.1).
6. a kind of fluidization for biological tissue according to claim 1 loads or takes out the device of cryoprotective agent,
It is characterized by: the solution loop module includes check valve (11), mixes container (5), the first peristaltic pump (7), the first heat exchange
Device (8) and tapping valve (10), the check valve (11) mix container (5), the first peristaltic pump (7), First Heat Exchanger (8) along molten
Liquid flow direction sequentially connects, and the First Heat Exchanger (8) is located in temperature control box (14), the inlet point of the tapping valve (10)
It is not connected with the inlet of the liquid outlet (9.5) of sample container (9) and check valve (11)；The solution loop module and sample
The inlet (9.4) of container (9) be connected with liquid outlet (9.5) First Heat Exchanger (8) for referring to solution loop module go out liquid
Mouthful, the inlet of check valve (11) is connected with the inlet (9.4) of sample container (9), liquid outlet (9.5) respectively；The control
Device (15) processed is electrically connected with solution loop module refers to that controller (15) is electrically connected with the first peristaltic pump (7).
7. a kind of fluidization for biological tissue according to claim 6 loads or takes out the device of cryoprotective agent,
It is characterized by: the solution supplying module includes the first fluid reservoir (1), the second fluid reservoir (2), the second peristaltic pump (3), third
Peristaltic pump (4), conductance electrode (6) and platinum resistance temperature sensor (16), first fluid reservoir (1) and the second peristaltic pump (3)
Inlet be connected, second fluid reservoir (2) is connected with the inlet of third peristaltic pump (4), the conductance electrode (6)
It is located at platinum resistance temperature sensor (16) and mixes in container (5)；The solution supplying module is connected with solution loop module
Refer to after the liquid outlet of the second peristaltic pump (3) is connected to the liquid outlet of third peristaltic pump (4) and to go out liquid with check valve (11) respectively
Mouth, the inlet for mixing container (5) are connected；The controller (15) is electrically connected with solution supplying module refers to controller (15)
It is electrically connected respectively with the second peristaltic pump (3), third peristaltic pump (4), conductance electrode (6) and platinum resistance temperature sensor (16).
8. a kind of fluidization for biological tissue according to claim 1 loads or takes out the device of cryoprotective agent,
It is characterized by: the gas supplying module includes gas generator (12), the second heat exchanger (13), gas generator (12) with
Second heat exchanger (13) is connected, and second heat exchanger (13) is located in temperature control box (14)；The gas supplying module and sample
The air inlet (9.6) of container (9) is connected the air inlet of the gas outlet Yu sample container (9) that refer to the second heat exchanger (13)
(9.6) it is connected；The controller (15) is electrically connected with gas supplying module refers to controller (15) and gas generator (12)
Electrical connection.
9. a kind of fluidization for biological tissue according to claim 8 loads or takes out the device of cryoprotective agent,
It is characterized by: the gas in the gas supplying module is nitrogen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610620317.4A CN106234351B (en) | 2016-07-29 | 2016-07-29 | For the fluidization load of biological tissue or the device of taking-up cryoprotective agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610620317.4A CN106234351B (en) | 2016-07-29 | 2016-07-29 | For the fluidization load of biological tissue or the device of taking-up cryoprotective agent |
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| CN106234351B true CN106234351B (en) | 2019-06-11 |
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| CN201610620317.4A Active CN106234351B (en) | 2016-07-29 | 2016-07-29 | For the fluidization load of biological tissue or the device of taking-up cryoprotective agent |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106680053A (en) * | 2017-01-04 | 2017-05-17 | 浙江大学 | Tissue treatment fluid circulation system |
| CN109628297B (en) * | 2018-12-14 | 2022-02-18 | 浙江大学宁波理工学院 | Microfluidic high-flux biological sample dripping and freezing storage device |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101173834A (en) * | 2007-09-28 | 2008-05-07 | 浙江大学 | Low temperature protectant solution program controlled refrigerating mechanism for vitrified preservation |
| CN104986446A (en) * | 2015-05-29 | 2015-10-21 | 浙江省医疗器械研究所 | Automated biological low temperature preservation device |
| CN105532638A (en) * | 2014-10-28 | 2016-05-04 | 无锡灵锡医疗器械科技有限公司 | Instrument apparatus for low-temperature storage of biological material |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101173834A (en) * | 2007-09-28 | 2008-05-07 | 浙江大学 | Low temperature protectant solution program controlled refrigerating mechanism for vitrified preservation |
| CN105532638A (en) * | 2014-10-28 | 2016-05-04 | 无锡灵锡医疗器械科技有限公司 | Instrument apparatus for low-temperature storage of biological material |
| CN104986446A (en) * | 2015-05-29 | 2015-10-21 | 浙江省医疗器械研究所 | Automated biological low temperature preservation device |
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