CN117229886A - Constant-temperature tissue digestion device and primary cell separation method - Google Patents
Constant-temperature tissue digestion device and primary cell separation method Download PDFInfo
- Publication number
- CN117229886A CN117229886A CN202311221173.1A CN202311221173A CN117229886A CN 117229886 A CN117229886 A CN 117229886A CN 202311221173 A CN202311221173 A CN 202311221173A CN 117229886 A CN117229886 A CN 117229886A
- Authority
- CN
- China
- Prior art keywords
- sample container
- rotating shaft
- sample
- connecting rod
- constant temperature
- 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
- 230000029087 digestion Effects 0.000 title claims abstract description 96
- 238000003320 cell separation method Methods 0.000 title claims abstract description 9
- 238000001514 detection method Methods 0.000 claims abstract description 85
- 230000001079 digestive effect Effects 0.000 claims abstract description 20
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 19
- 230000007246 mechanism Effects 0.000 claims abstract description 8
- 238000010494 dissociation reaction Methods 0.000 claims description 21
- 230000005593 dissociations Effects 0.000 claims description 21
- 230000000712 assembly Effects 0.000 claims description 7
- 238000000429 assembly Methods 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 239000002245 particle Substances 0.000 claims description 7
- 230000017525 heat dissipation Effects 0.000 claims description 6
- 230000001154 acute effect Effects 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 3
- 239000012780 transparent material Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 abstract description 9
- 210000001519 tissue Anatomy 0.000 description 69
- 238000000034 method Methods 0.000 description 18
- 230000006872 improvement Effects 0.000 description 12
- 238000010586 diagram Methods 0.000 description 11
- 230000000694 effects Effects 0.000 description 11
- 230000008569 process Effects 0.000 description 10
- 230000003628 erosive effect Effects 0.000 description 6
- 230000009471 action Effects 0.000 description 4
- 239000003570 air Substances 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 230000010355 oscillation Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000004364 calculation method Methods 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 102000038379 digestive enzymes Human genes 0.000 description 2
- 108091007734 digestive enzymes Proteins 0.000 description 2
- 239000004744 fabric Substances 0.000 description 2
- 230000005484 gravity Effects 0.000 description 2
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 102000016911 Deoxyribonucleases Human genes 0.000 description 1
- 108010053770 Deoxyribonucleases Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- -1 acryl Chemical group 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000012080 ambient air Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011423 initialization method Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention discloses a constant-temperature tissue digestion device and a primary cell separation method, wherein the constant-temperature tissue digestion device comprises a constant-temperature system for providing a constant-temperature environment, an oscillating mechanism for fully contacting a sample with a digestion liquid and a detection device for detecting the size of a cell mass; the vibration mechanism comprises a rotating shaft which is transversely arranged, and the rotating shaft can rotate; a plurality of connecting rods are radially arranged on the rotating shaft; the connecting rods are divided into a plurality of groups, and each group of connecting rods is arranged on the same circumference of the rotating shaft; the end part of the connecting rod is provided with a sample container clamp, and the angle between the sample container clamp and the connecting rod is adjustable; the sample container is clamped by the sample container clamp, and an included angle is formed between the sample container and the connecting rod; the invention abandons the existing plane reciprocating vibration table, changes the vibration mode into three-dimensional vibration, and can ensure that the digestive juice is fully contacted with the sample, thereby improving the digestion efficiency.
Description
Technical Field
The invention belongs to the technical field of primary cell culture, and particularly relates to a constant-temperature tissue digestion device and a primary cell separation method.
Background
Primary cell culture is increasingly used as a major tool in cell and molecular biology, providing an excellent model system for studying the normal physiology and biochemistry (e.g., metabolic studies, aging, signaling studies) of cells, the actions of drugs and toxic compounds. The current way to obtain primary cells from tissue is generally dissociation of various digestive enzymes. The digestive enzymes in common tissue digests are: trypsin, collagenase, DNase, etc., further loosen the bridge structure between cells by biochemical action of enzyme and chemical action of non-enzyme, make the block bulk, change the block form into flocculent, dissociate cells from tissues, and change into single cell or cell block state for subsequent in vitro culture.
Traditional enzyme digestion is mainly realized by means of a constant temperature tissue digestion device. The existing constant temperature tissue digestion device has the following defects: 1. when the tissue is subjected to enzyme digestion treatment, the sample is gathered at the bottom tip of the centrifugal tube, so that the sample is insufficiently contacted with digestion liquid, and the digestion efficiency is affected; 2. the optimal temperature of the enzymolysis reaction is 37 ℃, and the influence of the temperature on the enzyme activity is large, so that the digestion efficiency is influenced and the cell activity is influenced; 3. the traditional digestion mode mostly adopts a 37 ℃ water bath mode to control the temperature, in order to keep the digestion efficiency, the evenly mixed tissues and the digestion liquid need to be taken out for many times in the digestion process, the water bath mode is difficult to apply mechanical force to maintain the sufficient contact of the tissues and the digestion liquid for a long time, meanwhile, the water bath is easy to contact with a sample pipe orifice, and the risk of microbial contamination is increased; 4. the simple use of digestive juice to digest tissues has lower cell dissociation efficiency and longer digestion time, and the dissociated cells are soaked in the digestive juice for a long time, so that the activity of the cells is greatly influenced, and the success rate of the subsequent cell culture is reduced; 5. in the tissue dissociation process, the size of dissociated cell clusters can only be observed under a cell suspension mirror by an experimenter, so that the experimenter needs to observe under a liquid taking mirror for multiple times to control the digestion time to obtain the cell clusters with the target size, the operation time is increased, and the risk of microbial pollution is increased.
Disclosure of Invention
The invention aims to provide a constant-temperature tissue digestion device and a primary cell separation method, which can improve the digestion efficiency of a digestion solution on a sample.
In order to solve the technical problems, the invention adopts the following technical scheme: a constant temperature tissue digestion device, characterized in that: the device comprises a constant temperature system for providing a constant temperature environment, an oscillating mechanism for fully contacting a sample with digestive juice, and a detection device for detecting the size of cell clusters;
the constant temperature system comprises a closed shell, and a temperature sensor is arranged in the shell; the heating device is arranged in the shell, and the heat dissipation device is used for cooling air in the shell;
the vibration mechanism comprises a rotating shaft which is transversely arranged; at least one group of connecting rod assemblies are arranged on the rotating shaft at intervals along the axial direction; each group of connecting rod assemblies comprises a plurality of connecting rods which are arranged at intervals along the circumferential direction of the rotating shaft; the end part of the connecting rod is provided with a sample container clamp, and the angle between the sample container clamp and the connecting rod is adjustable; the sample container is clamped by a sample container clamp, so that the projection of the axis of the sample container on the plane of the connecting rod axis and the rotating shaft axis forms an acute angle alpha with the axis of the connecting rod;
the detection device is an ultrasonic particle size detection device and comprises an ultrasonic generator and an ultrasonic receiver which are arranged oppositely; a detection area is formed between the ultrasonic generator and the ultrasonic receiver, and the sample container can pass through the detection area when driven to rotate by the rotating shaft.
As an improvement, the axis of the sample container, the axis of the connecting rod and the axis of the rotating shaft are in the same plane.
As a further improvement, the acute angle is formed between the axis of the sample container and the plane in which the axis of the connecting rod and the axis of the rotating shaft lie.
As an improvement, the sample container is a cylindrical sleeve and a conical accommodating cavity is arranged at one end of the sleeve, and a screen is arranged in the sample container; the screen cloth is fixed in sleeve bottom, be provided with the crossbeam that is convenient for press from both sides in the sleeve.
As an improvement, the rotating shaft is provided with a clamping groove; the connecting ring is fixed on the rotating shaft by a clamping groove; the connecting rod is fixed on the connecting ring.
As an improvement, the same group of connecting rods are arranged on the same connecting ring and are uniformly distributed along the circumference of the connecting ring.
As an improvement, the heating device is a resistance wire, and the heat dissipation device is an exhaust fan.
As an improvement, the shell is made of transparent materials, and a door plate is arranged on the shell.
As an improvement, the rotating shaft is supported by the left bracket and the right bracket and is driven to rotate by the motor.
As an improvement, the device also comprises an initializing device, wherein the initializing device comprises a bulge which is arranged on the rotating shaft and is parallel to any connecting rod in the connecting rod assembly; the telescopic rod is arranged on the bracket, and can interfere with the bulge when the telescopic rod stretches out.
As another improvement, the initializing device comprises a protrusion extending along the radial direction on the rotating shaft, wherein the projection of the protrusion on the circumferential plane of the rotating shaft is positioned between the projections of the adjacent two connecting rods on the circumferential plane, and the initializing device further comprises a telescopic rod arranged on the bracket, and the telescopic rod can interfere with the protrusion when extending out, so that the initializing positioning is realized.
As an improvement, the motor control device further comprises a gear adjusting device, wherein the gear adjusting device can control the rotating speed of the motor.
The invention also provides a primary cell separation method which is applied to the constant temperature tissue digestion device and comprises the following steps:
placing a sample and a digestive juice in a sample container with a number;
connecting the sample containers with the connecting rods sequentially, wherein the serial numbers of the sample containers in the same group are arranged from small to large in the direction opposite to the rotating direction of the rotating shaft;
initializing the sample container positions such that the first-order sample containers in the same group are located within or near the detection zone of the detection device (also the first-order sample containers will be the first sample containers entering the detection zone); after the rotating shaft starts to rotate, sample containers in the same group from the first position to the last position sequentially pass through the detection area;
the constant temperature system is used for controlling the ambient temperature to be 36-38 ℃ and driving the rotating shaft to rotate for tissue digestion;
when different samples are placed in the sample containers, detecting the dissociation degree of the samples in each sample container by using a detection device; when the dissociation of the sample in a certain sample container reaches the target, stopping taking down the sample container with the corresponding number;
when the same sample is placed in the sample containers, detecting the dissociation degree of the sample in each sample container by using a detection device; and when the dissociated cell proportion reaches a first preset target, reducing the rotating speed of the rotating shaft until the dissociated cell proportion reaches a second preset target, and stopping the machine, wherein the first preset target is smaller than the second preset target.
As an improvement, the time interval of each sample container passing through the detection area is calculated by using the formula t=npi/180ω, where t is the time interval, n is the angle between the connecting rod where the currently detected sample container is located and the connecting rod where the first cis sample container is located in the same group, and ω is the angular velocity of the rotating shaft.
The invention has the advantages that:
firstly, the invention abandons the prior method of realizing plane rotation through a shaking table, changes the plane reciprocating vibration mode into three-dimensional rotation, and can lead the digestive juice to be fully contacted with the sample, thereby improving the digestion efficiency. In addition, undigested samples are separated from the cell mass by using a screen, so that the digestion implementation condition can be conveniently monitored by a detection device. The invention also adopts the heating and ventilation of the ambient air to ensure that the ambient temperature is constant at about 37 ℃, thereby avoiding the possibility of pollution of the prior water bath constant temperature to the mouth of the sample container.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. Like elements or portions are generally identified by like reference numerals throughout the several figures. In the drawings, elements or portions thereof are not necessarily drawn to scale. It will be apparent to those of ordinary skill in the art that the drawings in the following description are of some embodiments of the invention and that other drawings may be derived from these drawings without inventive faculty.
FIG. 1 is a schematic diagram of a isothermally digested tissue device in accordance with an exemplary embodiment of the present invention;
FIG. 2 is an enlarged view of the shaft of a constant temperature digestive tissue device according to an exemplary embodiment of the present invention;
FIGS. 3a to 3d are schematic views of the spindle in an initialized position in different initialized configurations of the apparatus for constant temperature digestion of tissue according to an exemplary embodiment of the present invention;
FIG. 4 is a schematic view of a sample container in a isothermally digested tissue device in accordance with an exemplary embodiment of the invention;
FIG. 5 is a schematic view of the angular relationship between the axis O3 of the sample container, the axis O2 of the connecting rod and the axis O1 of the spindle in the isothermally digested tissue device in accordance with an exemplary embodiment of the invention;
FIGS. 6a and 6b are schematic diagrams of cell clusters obtained during the same time period of digestion of the same batch of the same soft erosive tissue with a conventional tissue digestion unit and a tissue digestion unit according to the invention, respectively;
FIGS. 6c and 6d are schematic diagrams of cell clusters obtained during the same time period of digestion of a conventional tissue digestion unit and a tissue digestion unit of the present invention, respectively, with another batch of the same soft erosive tissue;
FIGS. 7a and 7b are schematic diagrams of cell clusters during the same time period of digestion with the same batch of the same tough tissue through a conventional tissue digestion device and a tissue digestion device of the present invention, respectively;
FIGS. 7c and 7d are schematic diagrams of a cell mass during the same time period of digestion with another batch of the same, more tough tissue through a conventional tissue digestion device and a tissue digestion device of the present invention, respectively;
FIGS. 8a and 8b are schematic views of a cell mass during the same time period of digestion with the same batch of the same hard tissue through a conventional tissue digestion device and a tissue digestion device of the present invention, respectively;
FIGS. 8c and 8d are schematic views of a cell mass during the same time period of digestion with another batch of the same hard tissue through a conventional tissue digestion device and a tissue digestion device of the present invention, respectively;
FIG. 9 is a table showing the comparison of the times required for the statistically obtained soft erosive tissue, tough tissue and hard tissue to achieve the same degree of dissociation using conventional digestion means and the digestion unit of the present invention, respectively.
Reference numerals: 1 casing, 2 sample container clamp, 3 door handle, 4 support, 5 ultrasonic receiver, 6 ultrasonic generator, 7 switch, 8 gear adjusting device, 9 detection device switch, 10 temperature display screen, 11 screen, 12 temperature adjustment button, 13 loose-leaf, 14 pivot, 15 connecting rod, 16 lid, 17 digestive juice, 18 sample, 19 screen cloth, 20 cell mass, 21 sample container, 22 sleeve, 23 crossbeam, 24 go-between, 25 detection area, 26 arch, 27 telescopic link.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. It will be apparent that the described embodiments are some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In this document, suffixes such as "module", "component", or "unit" used to represent elements are used only for facilitating the description of the present invention, and have no particular meaning in themselves. Thus, "module," "component," or "unit" may be used in combination.
The terms "upper," "lower," "inner," "outer," "front," "rear," "one end," "the other end," and the like herein refer to an orientation or positional relationship based on that shown in the drawings, merely for convenience of description and to simplify the description, and do not denote or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be construed as limiting the invention. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The terms "mounted," "configured to," "connected," and the like, herein, are to be construed broadly as, for example, "connected," whether fixedly, detachably, or integrally connected, unless otherwise specifically defined and limited; the two components can be mechanically connected, can be directly connected or can be indirectly connected through an intermediate medium, and can be communicated with each other. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art. Herein, "and/or" includes any and all combinations of one or more of the associated listed items. Herein, "plurality" means two or more, i.e., it includes two, three, four, five, etc.
Chinese patent CN201811323391.5 discloses a controllable device for the constant temperature digestion process of tissue samples, comprising: a housing on which a display screen is provided; the temperature control device is arranged in the shell and used for controlling the temperature in the shell, and the temperature control device displays the temperature value on the display screen in real time; the universal shaking table device is arranged in the shell, a sample container is arranged on the universal shaking table device, a tissue sample is placed in the sample container, and the universal shaking table device drives the sample container to shake; the sample detection device comprises an infrared emission device and an infrared receiving device, wherein the infrared emission device and the infrared receiving device are arranged at two sides of the sample container, infrared light emitted by the infrared emission device passes through a tissue sample in the sample container and then is converted into an electric signal by the infrared receiving device, and the electric signal is displayed on the display screen in real time.
In the prior art, the tray in the universal shaker device for uniformly mixing the sample and the digestive juice can rotate 360 degrees on the horizontal plane. However, in practical use, it is found that the sample still gathers in the sample container due to the centripetal force, and then rotates in a plane along with the digestive juice, that is, the sample and the digestive juice are rotated as a whole to form a unidirectional flow (or vortex), and the gathering phenomenon greatly reduces the contact area of the sample and the digestive juice, that is, the sample and the digestive juice are not sufficiently mixed, so that the digestion effect of the whole device is affected.
In addition, chinese patent CN202222869693.0 discloses a constant temperature sample incubator for cell therapy, including the incubator, be provided with the heater strip in the incubator, the heater strip sets up one side lateral wall of incubator, be provided with in the incubator and vibrate the module, the opposite side of incubator is provided with observation window.
The oscillation module in the prior art is used in the cell culture process, and has a main function of avoiding the problem that the oxygen supply capacity is poor and the cell proliferation is difficult to induce, so that the oxygen supply is increased in an oscillation mode to induce the cell proliferation. On the other hand, in the process of driving the storage cylinder to oscillate, the storage cylinder always keeps the vertical direction (namely, the axis of the storage cylinder performs circular motion in the plane of the motion path), and the storage cylinder is basically: the culture solution and cells in the storage cylinder still oscillate up and down in a vertical plane, so that dissolved oxygen is increased, and simultaneously, the cells are prevented from rotating on a culture surface in the oscillation process, and physical damage is caused to the cells. In addition, because the diameter of the storage cylinder is generally smaller and the axial direction is longer, the space for realizing uniform mixing of liquid after vertical placement is limited by forming rotational flow through up-and-down oscillation in the interior, and the effect of fully uniform mixing can not be achieved.
In order to solve the technical problem, as shown in fig. 1, the invention provides a constant temperature tissue digestion device, which comprises a constant temperature system for providing a constant temperature environment, an oscillating mechanism for fully contacting a sample with a digestion liquid, and a detection device for detecting the size of a cell mass (or sample tissue particles after the sample is digested).
In this embodiment, the constant temperature system comprises a closed casing 1, and a temperature sensor (not shown in the figure) is arranged in the casing 1; the air conditioner also comprises a heating device arranged in the shell 1 and a heat dissipation device for cooling air in the shell 1.
In this embodiment, the housing 1 is made of transparent material, such as glass or acryl. In order to achieve the heat preservation effect, a double-layer vacuum structure can be adopted. For easy installation or handling of the sample container 21, it is conceivable that a door panel which can be opened and closed is provided on the housing 1, which door panel is connected to the housing 1 via a hinge 13 and is provided with a door handle 3. The shell 1 is supported by a host machine, and other relevant components are installed on the host machine, and corresponding description will be made later.
The heating device and the heat dissipation device in the implementation are respectively selected from a resistance wire and an exhaust fan. The resistance wire is heated by air so that the ambient temperature in the whole housing 1 is kept around 37 ℃. When the temperature is too high, the exhaust fan can be started to ventilate the shell, so that the temperature is reduced.
In this embodiment, the vibration mechanism includes a rotating shaft 14 disposed transversely (i.e., when the entire device is placed horizontally, the axis of the rotating shaft is parallel to the horizontal plane), and the rotating shaft 14 is rotatable; a plurality of connecting rods 15 are radially arranged on the rotating shaft 14; the plurality of connecting rods 15 are divided into a plurality of groups, each group of connecting rods is arranged on the same circumference of the rotating shaft 14 (i.e. a plurality of groups of connecting rod assemblies are arranged along the axial direction at intervals, each group of connecting rod assemblies comprises a plurality of connecting rods extending along the radial direction of the rotating shaft, i.e. a plurality of connecting rods in the same group are uniformly arranged along the circumference of the rotating shaft at intervals, and the connecting rods can be arranged into 1 group or more than 1 group according to actual needs; the end part of the connecting rod 15 is provided with a sample container clamp 2, and the angle between the sample container clamp 2 and the connecting rod 15 can be adjusted; also included is a sample container 21 for placing a sample, the sample container 21 being held by the sample container holder 2 with the axis of the sample container 21 and the axis of the connecting rod 15 forming an acute angle α (see fig. 5). For example, the sample container clamp can be a test tube clamp or an O-shaped clamp, and the angle between the central line of the clamp and the axis of the connecting rod can be adjusted by adjusting the angle between the clamp and the connecting rod, so that the included angle alpha between the axis of the sample container, such as the axis of the test tube, and the axis of the connecting rod is realized.
In the present invention, the term "forming an angle" means that the axis of the sample container 21 and the axis of the connecting rod 15 are not aligned in parallel. Preferably, referring to fig. 5, the axis O3 of the sample container, the axis O2 of the connecting rod and the axis O1 of the rotating shaft lie in the same vertical plane (of course, the axis of the sample container may also form an angle with the plane in which the axis of the connecting rod and the axis of the rotating shaft lie). Further, the angle α between the axis O3 of the sample container 21 and the projection of the axis O2 of the connecting rod 15 in this vertical plane is 45 ° -75 ° (preferably 45 °), so that the sample container as a whole is disposed obliquely.
When the sample container 21 arranged obliquely rotates in the vertical plane, the sample container and the axis O1 of the rotating shaft are not arranged in parallel, or the axis of the sample container is not in the plane of the moving path of the sample container, but forms an included angle with the axis, so that in the rotating process, under the action of gravity, the sample and the digestive juice are continuously rolled (scattered due to rolling even if aggregation happens occasionally), thereby greatly increasing the contact area of the digestive juice and tissues, namely, a certain gravity effect is added compared with a universal shaking table mode, and the sample and the digestive juice are contacted and mixed thoroughly by not only shaking in the plane. When the included angle between the connecting rod 15 and the sample container 21 is 45-75 degrees, the mixing effect is more ideal; at the same time, it can be ensured that the receiving cavity at least at the bottom of the sample container can be completely placed in the detection area during the process of the connecting rod passing through the detection area, so that the cell mass in the receiving cavity is detected, and, even if two sample containers may exist in the detection area at the same time (for example, one sample container just enters the detection area and the last sample container just starts to leave the detection area) due to the fact that the bottom of the sample container is close to the rotating shaft side, only the bottom of the one sample container is located in the detection area.
While another reason for not choosing to place the sample container 21 horizontally is to avoid direct contact of the liquid with the nozzle and to reduce the risk of microbial contamination.
In order to facilitate the adjustment of the included angle between the sample container 21 and the connecting rod 15, the angle between the sample container clamp 2 and the connecting rod 15 in the invention can be adjusted, and different included angles can be adjusted according to different types of sample containers 21, so as to improve the mixing efficiency.
In this embodiment, the sample container clamp 2 is formed by an O-shaped pipe clamp, and a rubber pad is covered inside the sample container clamp 2, so as to fix, shock-proof and reduce abrasion of the sample container 21.
In this embodiment, the rotating shaft 14 is supported by the left and right brackets 4 and is rotated by a motor. The gear adjusting device 8 is further included, and the gear adjusting device 8 can control the rotating speed of the motor. For example, 5 gears 1-5 can be set, and different gears correspond to different motor speeds so as to cope with different use scenes (for example, different types of tissues to be digested). In this embodiment, the rotational speeds corresponding to 1 st to 5 th are 10rpm, 20rpm, 30rpm, 40rpm, and 50rpm, respectively.
More specifically, the rotating shaft 14 is provided with a clamping groove; the connecting ring 24 is also included, and the connecting ring 24 is fixed on the rotating shaft by a clamping groove; the connecting rod 15 is fixed on the connecting ring 24; the same group of connecting rods 15 are arranged on the same connecting ring 24 and are uniformly distributed along the circumference of the connecting ring 24. The connecting ring 24 is removable from the spindle 14 to facilitate the integral replacement of the sample container 21 on the spindle 14. In this embodiment, the connecting rings 24 are arranged in front and rear along the axial direction of the rotating shaft 14, so there are two groups of connecting rods 15. Of course, the number of connection rings 24 may be set as desired. Likewise, the number of connecting rods 15 may be arranged according to the requirements or the size. In this embodiment, 6 connecting rods 15 are provided, and the included angle between the connecting rods 15 is 60 °.
In addition, the bottom of the sample container 21 is conical (i.e. a conical accommodating cavity is formed for accommodating the cell mass obtained by filtration), and a screen 19 is arranged in the sample container 21; specifically, the screen 19 is fixed to the bottom of the sleeve 22, and a cross beam 23 for facilitating gripping is provided in the sleeve 22. The mesh 19 has a pore size of 40um to 150um and is supported by the sleeve 22. The screen 19 and sleeve 22 are individually packaged for disposable use after sterilization. Of course, the sample container 21 is also provided with a cover 16 to avoid spillage. When the device is used, the sleeve 22 is inserted into the sample container 21, then the sample 18 to be digested and the digestion liquid 17 are put into the sleeve, the cell mass 20 dissociated through the rotary digestion and enzyme digestion effects enters the conical accommodating cavity of the sample container 21 through the screening of the screen 19, the cell mass 20 at the bottom is monitored in real time by the detection device, when the ratio of the cell mass 20 at the bottom reaches a preset value, the digestion can be stopped, the sleeve 22 with the screen 19 is taken out by using sterile forceps, and the rapid separation of the target cell mass at the bottom, the non-target sample at the upper layer and impurities is realized.
The detection device is an ultrasonic particle size detection device and comprises an ultrasonic generator 6 and an ultrasonic receiver 5 which are arranged oppositely; a detection area 25 is formed between the ultrasonic generator 6 and the ultrasonic receiver 5, and the sample container 21 can pass through the detection area 25 when being driven to rotate by the rotating shaft 14.
In this embodiment, since two sets of sample containers 21 are provided, two sets of ultrasonic particle size detection devices are also required. For example, the ultrasonic generator 6 is arranged at the top of the housing 1, and the ultrasonic receiver 5 is arranged at the corresponding position of the base, so that a detection area 25 is formed between the ultrasonic generator and the base. When the sample container 21 rotates into the detection area 25, the difference of the absorption degree of the ultrasonic wave by the particles with different sizes makes the attenuation degree of the ultrasonic wave obtained by the ultrasonic receiver 5 end have a difference, so that the main control module of the device can judge the size of the cell mass (or tissue particles in the accommodating cavity) according to the signal fed back by the ultrasonic receiver, and analyze the duty ratio of the target cell mass so as to prompt the digestion progress degree.
In addition, since the sample container 21 is rotated in the present invention, a problem to be solved in the detection is how to judge which sample container is currently detected. The method employed in the present invention is judged by taking the time interval at which each sample container 21 passes through the ultrasonic device, that is, since the relative position between the sample containers 21 is fixed and the rotation speed of the rotation shaft is known, it is only necessary to determine the initial position, i.e., which sample container 21 is located in or near the detection area at the start of operation.
In order to solve the problem of initialization, the present invention provides an initializing device, see fig. 3a and 3b, which includes a protrusion 26 extending radially along the rotation axis and parallel to one of the connecting rods 15 in the group of connecting rods 15; also comprises a telescopic rod 27 arranged on the bracket 4, and can interfere with the bulge 26 when the telescopic rod 27 stretches out. Thus, when the position at which the expansion link 27 interferes with the boss 26 to prevent the rotation of the rotation shaft 14 is set as the initial position, at this time, the axis of the boss is parallel to the center line (i.e., the vertical direction) of the detection area.
Referring to fig. 3a, when the rotating shaft 14 rotates clockwise, one of the two sample containers close to the detection area, which is far away from the corresponding connection rod of the protrusion, is the initial sample container (i.e. the first sequential position in the same group, such as No. 1 or No. 7); referring to fig. 3b, when the rotating shaft 14 rotates counterclockwise, one of the two sample containers adjacent to the detection area, which is adjacent to the connecting rod corresponding to the protrusion, is the initial sample container (i.e., the first sequential position in the same group, such as No. 1 or No. 7).
In other embodiments, see fig. 3c, the initializing device comprises a protrusion 26 extending radially along the rotation axis 14 and located between two adjacent connection bars 15 (located between two connection bars means that the projection of the protrusion on the circumferential plane of the rotation axis is located between the projections of the two connection bars on the circumferential plane), and a telescopic bar 27 arranged on the support 4, which may interfere with the protrusion 26 when the telescopic bar 27 is extended (of course, further, in order to enable the control system of the device to automatically recognize, a pressure sensor may be arranged on the protrusion or the telescopic bar, so that when the two interfere, a corresponding signal may be fed back to the control system, which may control to stop the rotation and know that the initialization is completed). Thus, the position when the telescopic rod 27 interferes with the projection 26 to prevent the rotation of the rotation shaft 14 is set as the initial position, where the axis of the projection is parallel to the center line of the detection area, and the sample container 21 located in the detection area 25 at this time is taken as the initial sample container 21 (for example, the first-order-numbered sample container in the same group). Thus, it is possible to know which sample container 21 the sample container 21 having entered the detection area 25 at each moment is by calculation.
In other embodiments, see fig. 3d, the initializing device comprises a protrusion 26 extending radially along the rotation axis 14 and parallel to the Ren Yilian extension bar 15, and a telescopic bar 27 arranged on the support 4, which may interfere with the protrusion 26 when the telescopic bar 27 is extended. Thus, when the position where the telescopic rod 27 interferes with the projection 26 to prevent the rotation of the rotation shaft 14 is set as the initial position, at this time, the axis of the projection is perpendicular to the center line of the detection area, and the sample container 21 located in the detection area 25 at this time is taken as the initial sample container 21 (for example, the first-order-numbered sample container in the same group). Thus, it is possible to know which sample container 21 is the sample container 21 entering the detection area 25 at the corresponding timing by estimation.
Specifically, referring to fig. 3d, in the initial state, the time interval when each sample container passes through the detection area can be calculated by using the formula t=npi/180ω, so that the number of the currently detected sample container can be calculated (for example, from the initial state, every t, 1 is accumulated from the initial number until the last number of the same group is restarted), where t is the time interval, n is the angle between the connecting rod where the currently detected sample container is located and the connecting rod where the first cis sample container is located in the same group, and ω is the angular velocity of the rotating shaft.
In addition, a power switch 7 is arranged on the host machine for controlling the power on/off of the whole device. A screen 11 is also provided for real-time display of the detected cell fraction in the sample container. A temperature adjusting button 12 for controlling the temperature and a temperature display screen 10 for displaying the current temperature are also provided. For aesthetic purposes, the motor for driving the shaft 14 may be disposed in the main machine and driven by gears or chains.
The invention also provides a primary cell separation method which is applied to the constant temperature tissue digestion device and comprises the following specific steps:
s1, placing a sample to be digested and digestive juice in a sample container with a number. The sleeve is placed in the sample container, and then a suspension of the prepared sample and digestive juice is injected into the sample container. Each sample container has an independent number, and in this embodiment, the number of 12 sample containers in total is 1-12 in the 2 groups, wherein 1 group is 1-6 sample containers, and 2 group is 7-12 sample containers.
S2, connecting the sample containers with the connecting rod sequentially, wherein the sequence number of the sample containers is arranged from small to large in the direction opposite to the rotating direction of the rotating shaft.
For example, the rotating shaft rotates clockwise, so that the sample containers are arranged in the order of the numbers from small to large in the anticlockwise direction, and the sample containers are arranged in the order when being detected, thereby facilitating later calculation. Of course, if the spindle is rotated counterclockwise, the sample containers are correspondingly mounted clockwise with the numbering being arranged in a sequence from small to large.
S3, initializing the sample container position so that one sample container of the same group of first order numbers (i.e., the smallest number in the same group) is located in or near the detection area of the detection device.
The initialization is to determine which sample container is located in the detection area when the operation is started. Of course, a convenient method is to place the first group of sample containers with the number 1 and the second group of sample containers with the number 7 in the detection area first, and after the rotation of the rotating shaft is started, the sample containers in the same group enter the detection area from the first position to the last position in sequence.
Of course, in other embodiments, referring to fig. 3a and 3b, when the positions of the sample containers are initialized, the two sample containers in the same group with the first sequential number (i.e. the smallest number in the same group) and the last position may be made to correspond to or be close to the detection area of the detection device (i.e. no sample container is currently located in the detection area), and when the spindle starts to rotate, the sample containers enter the detection area sequentially from the first sequential position to the last position (i.e. the distance L between the edge of the detection area and the spindle is smaller than the length R of the connecting rod).
The invention provides two initialization methods.
The first is manual. The vertical line can be marked on the bracket, so that when the connecting rods of the sample containers No. 5 and No. 11 are overlapped with the vertical line, the sample containers No. 1 and No. 7 are just positioned in the detection area.
The second is automatic, and the initialization is performed by the above-mentioned initializing device. Similarly, the connecting rod parallel protrusions where the sample containers No. 5 and No. 11 are located are arranged on the rotating shaft for interference. When the rotation of the rotation shaft is stopped, the sample container 1 and the sample container 7 are just located in the detection area (see fig. 3c and 3 b), or are close to the detection area (see fig. 3a and 3 b), and after the rotation shaft starts to rotate, the sample containers from the first position to the last position in the same group of sample containers sequentially pass through the detection area.
In some embodiments, when a user touches a corresponding initialization switch, the main control module of the constant temperature tissue digestion device triggers the driving device to drive the rotating shaft to rotate; when the bulge and the telescopic rod interfere, a pressure sensor arranged on the telescopic rod or the bulge feeds back a corresponding pressure signal to the main control module, and then the main control module controls the driving device to stop rotating and prompts that the initialization is completed; when a user touches a corresponding switch for starting tissue digestion, the main control module controls the driving device to drive the rotating shaft to rotate again, and simultaneously controls the detection device to start detecting (real-time detection or periodic detection) the dissociation degree (or digestion degree) of the sample to be digested in each sample container; and when the sample containers from the first order to the last order in the same group sequentially pass through the detection area, the detection device feeds back corresponding trigger signals to the main control module, and then the main control module calculates the number of the sample container currently positioned in the detection area and prompts the number of the sample container and the dissociation degree of tissues to be digested in the sample container.
For example, referring to fig. 3d, in an initial state, the main control module assigns a number identifying the sample container currently located in the detection area to an initial value, and adds 1 to the number N every Δt=α/w until the value of the number N is equal to the last number in the same group; and when the detection means detects that the next sample container is located in the detection zone, the number N is reinitialized.
For another example, when the main control module receives a trigger signal fed back by the detection device for the first time (the trigger signal is generated when the detection device detects that a sample container enters the detection area for the first time), the sample container number N which is marked as currently located in the detection area is given an initial value, and the number N is added by 1 every Δt=α/w until the value of the number N is equal to the last number in the same group; when the detection device detects that the next sample container is positioned in the detection area, the number N is initialized again; wherein, the initial value of the number N is the number of the first-order sample container in the same group of sample containers; alpha is the included angle between two adjacent connecting rods.
S4, controlling the ambient temperature to be 36-38 ℃ by utilizing a constant temperature system, and driving the rotating shaft to rotate to digest tissues. The invention can be applied to at least the following two working conditions.
Firstly, when different samples (the different samples have little difference in the requirements on the rotating speed) are placed in all the sample containers, the dissociation degree of the samples in each sample container is detected by using a detection device; when detecting that the dissociation of the sample in a certain sample container reaches the target, stopping and taking down the sample container with the corresponding number; digestion is then continued until all sample containers are removed. Since the various samples are simultaneously digested, the state of each sample is different, the degree of dissociation of the tissue sample within each sample container is monitored in real time or periodically.
Secondly, when the same samples (namely, a large number of tissue samples obtained by the same treatment mode are subjected to high-flux dissociation) are placed in all the sample containers, the dissociation degree of the samples in each sample container is detected by using a detection device (preferably, the dissociation degree of the samples in any sample container is periodically detected; because the tissue samples are the same and other conditions are the same, the dissociation degree in each centrifuge tube is not much different and can be ignored, so that the dissociation monitoring result of any sample container can be periodically monitored as the dissociation result of the group); when the dissociated cell proportion reaches a first preset target value, for example 45-55%, the rotating speed of the rotating shaft is reduced until the dissociated cell proportion reaches a second preset target value, for example 70-80%, the machine is stopped, and the first preset target value is relatively close to the second preset target value, so that the target cell mass is not excessively dissociated into single cells, the success rate of culture is improved, and the mechanical force effect is reduced by reducing the rotating speed.
For samples with different properties, the initial gear can be selected to be different, for example, 1-2 gears are selected for soft erosion type tissues, 3-4 gears are selected for tissues with stronger toughness of multi-fiber connective tissues, and 5 gears are selected for texture hard tissues.
Referring to fig. 9, after a large amount of statistics of experimental data, the effect of primary cell separation using the constant temperature tissue digestion device provided by the present invention is compared with that of the prior art: after primary cell separation is carried out by adopting the constant-temperature tissue digestion device provided by the invention, the length of the digestion time is reduced by more than 40%, and the efficiency is greatly improved.
Specifically, during the test, it was found that:
referring to fig. 6a and 6b, schematic diagrams of cell clusters obtained after the same soft erosion tissue is digested for the same time by adopting a conventional digestion mode and the digestion device of the invention respectively; referring to fig. 6c and 6d, the schematic diagrams of the cell mass obtained after the same time of digestion of another batch of the same soft erosion tissue by the conventional digestion method and the digestion device of the present invention show that the total amount of cells obtained by the digestion device of the present invention is larger after the digestion for the same time.
Referring to fig. 7a and 7b, schematic cell diagrams are shown obtained after the same tissue with stronger toughness is digested for the same time by adopting a conventional digestion mode and the digestion device of the invention; referring to fig. 7c and 7d, a schematic diagram of a cell mass obtained after digesting another group of more tough tissues for the same time using a conventional digestion method (as a control group) and a digestion device according to the present invention (as an experimental group), respectively. As is clear from the figure, the total amount of cells obtained by the digestion apparatus of the present invention was greater after the same time of digestion.
Referring to fig. 8a and 8b, schematic diagrams of cell clusters obtained after the tissue with higher toughness is digested for the same time by adopting a conventional digestion mode and the digestion device of the invention; referring to fig. 8c and 8d, a schematic diagram of a cell mass obtained after another batch of the same tough tissue was digested for the same time by using the conventional digestion means and the digestion device of the present invention, respectively. As is clear from the figure, the total amount of cells obtained by the digestion apparatus of the present invention was greater after the same time of digestion.
It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element.
The embodiments of the present invention have been described above with reference to the accompanying drawings, but the present invention is not limited to the above-described embodiments, which are merely illustrative and not restrictive, and many forms may be made by those having ordinary skill in the art without departing from the spirit of the present invention and the scope of the claims, which are to be protected by the present invention.
Claims (10)
1. A constant temperature tissue digestion device, characterized in that: the device comprises a constant temperature system for providing a constant temperature environment, an oscillating mechanism for fully contacting a sample with digestive juice, and a detection device for detecting the size of cell clusters;
the constant temperature system comprises a closed shell, and a temperature sensor is arranged in the shell; the heating device is arranged in the shell, and the heat dissipation device is used for cooling air in the shell;
the vibration mechanism comprises a rotating shaft which is transversely arranged; at least one group of connecting rod assemblies are arranged on the rotating shaft at intervals along the axial direction; each group of connecting rod assemblies comprises a plurality of connecting rods which are arranged at intervals along the circumferential direction of the rotating shaft and extend along the radial direction of the rotating shaft; the end part of the connecting rod is provided with a sample container clamp, and the angle between the sample container clamp and the connecting rod is adjustable; the sample container is clamped by the sample container clamp, so that the projection of the axis of the sample container on the plane of the connecting rod axis and the rotating shaft axis forms an acute angle alpha with the axis of the connecting rod;
the detection device is an ultrasonic particle size detection device and comprises an ultrasonic generator and an ultrasonic receiver which are arranged oppositely; a detection area is formed between the ultrasonic generator and the ultrasonic receiver, and the sample container can pass through the detection area when driven by the rotating shaft to rotate.
2. A constant temperature tissue digestion device as in claim 1, wherein: the axis of the sample container forms the acute angle with the axis of the connecting rod and the plane of the axis of the rotating shaft.
3. A constant temperature tissue digestion device as in claim 1, wherein: the sample container comprises a cylindrical sleeve and a conical accommodating cavity arranged at one end of the sleeve, and a screen is arranged in the sample container; the screen is fixed at the bottom of the sleeve, and a cross beam which is convenient to clamp is arranged in the sleeve; and/or the number of the groups of groups,
a clamping groove is formed in the rotating shaft; the connecting ring is fixed on the rotating shaft by the clamping groove; the connecting rod is fixed on the connecting ring; the same group of connecting rod assemblies are arranged on the same connecting ring and are uniformly distributed along the circumference of the connecting ring.
4. A constant temperature tissue digestion device as in claim 1, wherein: the heating device is a resistance wire, and the heat dissipation device is an exhaust fan.
5. A constant temperature tissue digestion device as in claim 1, wherein: the shell is made of transparent materials, and a door plate is arranged on the shell.
6. A constant temperature tissue digestion device as in claim 1, wherein: the rotating shaft is supported by the left bracket and the right bracket and driven by the motor to rotate.
7. A constant temperature tissue digestion device as in claim 6, wherein: the gear adjusting device can control the rotating speed of the motor.
8. A constant temperature tissue digestion device as in claim 6, wherein: the initializing device comprises a bulge which is arranged on the rotating shaft and is parallel to any connecting rod in the connecting rod assembly; the telescopic rod is arranged on the bracket, and can interfere with the bulge when extending out, so that the initialization positioning is realized; or, the initializing device comprises a bulge which is arranged on the rotating shaft and extends along the radial direction, the projection of the bulge on the circumferential plane of the rotating shaft is positioned between the projections of the adjacent two connecting rods on the circumferential plane, and the initializing device also comprises a telescopic rod which is arranged on the bracket, and when the telescopic rod stretches out, the telescopic rod can interfere with the bulge, so that the initializing positioning is realized.
9. A primary cell separation method applied to the constant temperature tissue digestion device according to any one of claims 1 to 8, characterized by comprising:
placing a sample to be digested and a digestive juice in a sample container with a number;
connecting the sample containers sequentially with connecting rods in the constant temperature tissue digestion device, wherein the serial numbers of the sample containers are arranged from small to large in the direction opposite to the rotating direction of a rotating shaft in the constant temperature tissue digestion device;
initializing the positions of the sample containers so that the sample containers numbered as the first order in the same group are positioned in or close to a detection area of a detection device in the constant temperature tissue digestion device; after the rotating shaft starts to rotate, sample containers numbered from the first position to the last position in the same group of sample containers sequentially pass through the detection area;
the constant temperature system is used for controlling the ambient temperature to be 36-38 ℃ and driving the rotating shaft to rotate for tissue digestion;
when different samples are placed in all the sample containers, detecting the dissociation degree of the samples in each sample container by using a detection device; when the dissociation of the sample in a certain sample container reaches the target, stopping taking down the sample container with the corresponding number;
when the same sample is placed in all the sample containers, detecting the dissociation degree of the sample in each sample container by using a detection device; and when the dissociated cell proportion reaches a first preset target value, reducing the rotating speed of the rotating shaft until the dissociated cell proportion reaches a second preset target value, and stopping the machine, wherein the first preset target value is smaller than the second preset target value.
10. The primary cell separation method of claim 9, wherein:
calculating the time interval of each sample container passing through the detection area by using the formula t=npi/180 omega, wherein t is the time interval, n is the included angle between the connecting rod where the currently detected sample container is positioned and the connecting rod where the first cis sample container is positioned in the same group, and omega is the angular speed of the rotating shaft.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311221173.1A CN117229886A (en) | 2023-09-20 | 2023-09-20 | Constant-temperature tissue digestion device and primary cell separation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311221173.1A CN117229886A (en) | 2023-09-20 | 2023-09-20 | Constant-temperature tissue digestion device and primary cell separation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117229886A true CN117229886A (en) | 2023-12-15 |
Family
ID=89083971
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311221173.1A Pending CN117229886A (en) | 2023-09-20 | 2023-09-20 | Constant-temperature tissue digestion device and primary cell separation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117229886A (en) |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101733038A (en) * | 2008-11-25 | 2010-06-16 | 李雪兴 | rotary incubator |
| DE102009007822A1 (en) * | 2009-02-07 | 2010-08-19 | PRO DESIGN Gesellschaft für Produktentwicklung mbH | Shaking or vibrating device for producing flow disturbance in sample in laboratory during e.g. mixing of liquid, has holder, where container rotates with holder with respect to center axis that rotates about vertical axis on conical surface |
| CN203745269U (en) * | 2013-12-20 | 2014-07-30 | 深圳华大基因研究院 | Blending device |
| CN109298730A (en) * | 2018-11-07 | 2019-02-01 | 上海坤爱生物科技股份有限公司 | A kind of controllable device of tissue sample constant temperature digestion process |
| CN210834417U (en) * | 2019-10-16 | 2020-06-23 | 山东省食品药品检验研究院 | Bottled liquid food mixing device |
| WO2020135128A1 (en) * | 2018-12-28 | 2020-07-02 | 深圳迈瑞生物医疗电子股份有限公司 | Sample mixing apparatus, sample analysis system, and sample mixing method |
| CN211487407U (en) * | 2019-12-13 | 2020-09-15 | 山东省齐鲁干细胞工程有限公司 | Reagent spiral blending machine |
| CN215288767U (en) * | 2021-04-06 | 2021-12-24 | 深圳市第二人民医院(深圳市转化医学研究院) | A vibrate device for exosome draws |
| CN114682135A (en) * | 2020-12-28 | 2022-07-01 | 南京金斯瑞生物科技有限公司 | Turntable rotating device with uniformly mixing function station and use method thereof |
| CN115976152A (en) * | 2022-12-23 | 2023-04-18 | 四川大学 | Method of using centrifugal device for tissue digestion |
-
2023
- 2023-09-20 CN CN202311221173.1A patent/CN117229886A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101733038A (en) * | 2008-11-25 | 2010-06-16 | 李雪兴 | rotary incubator |
| DE102009007822A1 (en) * | 2009-02-07 | 2010-08-19 | PRO DESIGN Gesellschaft für Produktentwicklung mbH | Shaking or vibrating device for producing flow disturbance in sample in laboratory during e.g. mixing of liquid, has holder, where container rotates with holder with respect to center axis that rotates about vertical axis on conical surface |
| CN203745269U (en) * | 2013-12-20 | 2014-07-30 | 深圳华大基因研究院 | Blending device |
| CN109298730A (en) * | 2018-11-07 | 2019-02-01 | 上海坤爱生物科技股份有限公司 | A kind of controllable device of tissue sample constant temperature digestion process |
| WO2020135128A1 (en) * | 2018-12-28 | 2020-07-02 | 深圳迈瑞生物医疗电子股份有限公司 | Sample mixing apparatus, sample analysis system, and sample mixing method |
| CN210834417U (en) * | 2019-10-16 | 2020-06-23 | 山东省食品药品检验研究院 | Bottled liquid food mixing device |
| CN211487407U (en) * | 2019-12-13 | 2020-09-15 | 山东省齐鲁干细胞工程有限公司 | Reagent spiral blending machine |
| CN114682135A (en) * | 2020-12-28 | 2022-07-01 | 南京金斯瑞生物科技有限公司 | Turntable rotating device with uniformly mixing function station and use method thereof |
| CN215288767U (en) * | 2021-04-06 | 2021-12-24 | 深圳市第二人民医院(深圳市转化医学研究院) | A vibrate device for exosome draws |
| CN115976152A (en) * | 2022-12-23 | 2023-04-18 | 四川大学 | Method of using centrifugal device for tissue digestion |
Non-Patent Citations (1)
| Title |
|---|
| 杨松荣 北京：冶金工业出版社: "碎磨工艺及应用", 31 January 2013, pages: 15 - 16 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111378563B (en) | System and method for probiotic culture | |
| US5360741A (en) | DNA hybridization incubator | |
| CN108325932A (en) | A kind of full-automatic rotating thin film oven Sheng sample bottle cleaning device | |
| CN117229886A (en) | Constant-temperature tissue digestion device and primary cell separation method | |
| CN111744869A (en) | Cleaning and maintaining device for medical instrument | |
| CN108753614A (en) | Anti-tumor experiment tumor cell culture device | |
| CN112779132A (en) | Cell culture base full-automatic fermentation cylinder that disinfects evenly | |
| CN208341249U (en) | A kind of full-automatic rotating thin film oven Sheng sample bottle cleaning device | |
| CN210711527U (en) | Novel multi-functional microbial cultivation device | |
| CN209530711U (en) | A kind of intelligence oscillation vortex mixer | |
| CN111495615A (en) | Ozone ultraviolet composite disinfection centrifugal machine | |
| CN209010520U (en) | Adjustable speed constant temperature tissue dissociation device | |
| CN212397031U (en) | Ozone ultraviolet composite disinfection centrifugal machine | |
| CN211785159U (en) | Multichannel food short-term test appearance | |
| CN211071003U (en) | Slide glass belt cleaning device for microbiological experiments | |
| CN209974791U (en) | Glass fermentation tank | |
| CN106771116B (en) | A kind of strainer antianaphylaxis detection method and device | |
| CN207351087U (en) | A kind of Sterile vacuum drying box | |
| CN210367691U (en) | Staphylococcus aureus detection device | |
| CN220183228U (en) | Microorganism inspection extraction device | |
| JP2004236547A (en) | Cell culture disk and cell culture apparatus using the same | |
| CN218452135U (en) | Novel culture medium mixing equipment | |
| CN214132229U (en) | Vertical high-capacity aerosol purification centrifuge | |
| CN217385464U (en) | Waste water sample detection device for environmental detection | |
| CN219291458U (en) | Numerical control water bath |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Country or region after: China Address after: 610000 No. 701, 702, 704, Floor 7, Unit 2, Building 1, No. 818, Xingshi Road, Wuhou District, Chengdu, Sichuan Applicant after: Chengdu nuoyeide medical laboratory Co.,Ltd. Applicant after: Shenzhen Jingke Biotechnology Co.,Ltd. Address before: 610043 No. 701, 702, 704, 7th Floor, Unit 2, Building 1, No. 818, Xingshi Road, Wuhou District, Chengdu, Sichuan Province Applicant before: Chengdu nuoyeide medical laboratory Co.,Ltd. Country or region before: China Applicant before: GUAGNZHOU JINGKE BIOTECH CO.,LTD. |