CN212008339U - Imaging platform adaptive to tissue transparentization and in-vitro brain slice - Google Patents
Imaging platform adaptive to tissue transparentization and in-vitro brain slice Download PDFInfo
- Publication number
- CN212008339U CN212008339U CN202020241541.4U CN202020241541U CN212008339U CN 212008339 U CN212008339 U CN 212008339U CN 202020241541 U CN202020241541 U CN 202020241541U CN 212008339 U CN212008339 U CN 212008339U
- Authority
- CN
- China
- Prior art keywords
- platform
- observation
- inlet
- cerebrospinal fluid
- imaging platform
- 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.)
- Active
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The utility model discloses an imaging platform of adaptation tissue transparentization and separation brain piece, including platform main part, constant temperature equipment and artifical cerebrospinal fluid circulation mechanism, wherein the platform main part includes cavity body and observation platform, and the middle part of the bottom plate of observation platform is equipped with the observation groove that is used for placing separation brain piece, and artifical cerebrospinal fluid circulation mechanism communicates with the observation groove for the circulation of artifical cerebrospinal fluid in the observation groove can be controlled, has guaranteed to observe the circulation balance of artifical cerebrospinal fluid in the observation groove, ensures that separation brain piece keeps the activity, and then improves the imaging effect; simultaneously, the constant temperature equipment sets up in cavity originally internally, makes the temperature of platform main part can be controlled and keep under normal physiological temperature condition, guarantees separation brain piece survival rate to the utmost, ensures separation brain piece internal neuron vitality, and the experiment accuracy obtains very big improvement, and this utility model is used for biological tissue experiment field.
Description
Technical Field
The utility model relates to a biological tissue experiment field especially relates to an imaging platform of adaptation tissue transparentization and separation brain piece.
Background
The brain slice is used as a tissue specimen commonly used in neuroscience research, the brain tissue of an experimental animal is rapidly taken out, the brain tissue is fixed in agarose, a vibrating microtome is used for cutting into slices of 200-500 mu m in artificial cerebrospinal fluid, the slices retain the activity and the function of part of the nerve tissue, the advantages of relatively complete nerve circuits and extracellular environments are partially retained, the environment of in-vivo neurons can be simulated, the detection of the shape, the structure and the function of related neurons can be monitored in real time, and an important research means is provided for neuroscience research.
In recent years, based on the combination of a transparentization method with transgenic protein labeling, virus transfection and immune tissue fluorescent staining, by using imaging devices such as a confocal microscope, a multiphoton microscope and a laser scanning microscope, the fluorescent signal tracking after transparentization can be performed on nerve tissues with different thicknesses, and the research on the three-dimensional structure of biological tissues, the tracing of nerve loops, the distribution of specific cells in tissues and the like is gradually promoted.
At present, in colleges and universities and scientific research institutions, especially in related professions such as medicine and pharmacy, a large amount of experimental research related to nervous system activities is needed. The simple device made by the slide groove is usually used when the brain slice is detected in vitro and observed by tissue transparentization for a long time, and the simple device made by the user usually has the following problems: firstly, the speed of cerebrospinal fluid circulation cannot be controlled, and the liquid level is possibly too high or too low, so that the brain slice is unstable, and the imaging effect is influenced; the temperature cannot be stably controlled, so that the survival rate of the brain slices is influenced when the brain slices are exposed to room temperature for a long time, the functions and the biological activities of the intrinsic neurons are irreversibly influenced, and the accuracy of an experiment is seriously influenced; and thirdly, the repeatability is not high, the research cannot be repeated in other scientific research personnel laboratories, and the experimental credibility is limited.
SUMMERY OF THE UTILITY MODEL
An object of the utility model is to solve one of the technical problem that exists among the prior art at least, provide one kind and can improve the separation brain piece survival rate and improve the transparent and the imaging platform of separation brain piece of adaptation tissue of experiment accuracy.
The imaging platform comprises a platform main body, wherein the platform main body comprises a hollow body and an observation platform fixed above the hollow body, the observation platform comprises a bottom plate and a side wall plate, and the middle part of the bottom plate is provided with an observation groove for placing an in-vitro brain slice; the constant temperature device is arranged in the hollow body; and the artificial cerebrospinal fluid circulating mechanism is arranged on the hollow body and is communicated with the observation groove.
According to the utility model discloses imaging platform of adaptation tissue transparentization and separation brain piece possesses following beneficial effect at least: the imaging platform adaptive to tissue transparentization and in-vitro brain slices comprises a platform main body, a constant temperature device and an artificial cerebrospinal fluid circulating mechanism, wherein the platform main body comprises a hollow body and an observation table, an observation groove for placing the in-vitro brain slices is formed in the middle of a bottom plate of the observation table, and the artificial cerebrospinal fluid circulating mechanism is communicated with the observation groove, so that the circulation of artificial cerebrospinal fluid in the observation groove can be controlled, the circulation balance of the artificial cerebrospinal fluid in the observation groove is ensured to be observed, the in-vitro brain slices are ensured to keep activity, and the imaging effect is further improved; simultaneously, the constant temperature equipment sets up in this internally at cavity, makes the temperature of platform main part can be controlled and keep under normal physiological temperature condition, guarantees separation brain piece survival rate to the at utmost, ensures separation brain piece inside neuron vitality, and the experiment accuracy obtains very big improvement.
According to the utility model discloses a some embodiments, the middle part of bottom plate is provided with the column arch, it is located the bellied intermediate position of column to observe the trench, first inlet and first liquid outlet have been seted up to the bellied top surface of column, it is equipped with second inlet and second liquid outlet from bottom to top to observe the inslot.
According to some embodiments of the utility model, artifical cerebrospinal fluid circulation mechanism includes feed liquor pipe and drain pipe, first inlet and second inlet, first liquid outlet and second liquid outlet are respectively through feed liquor pipe, drain pipe switch-on.
According to some embodiments of the utility model, first inlet and first liquid outlet are respectively through hose connection to a peristaltic pump, two the peristaltic pump all is connected to in an artificial cerebrospinal fluid storage device, the outer wall of side wall board is equipped with the buckle with fixed hose.
According to the utility model discloses a some embodiments, form the accumulator between protruding and the side wall board of column, the bellied side of column inwards caves in and forms at least one and spills over the passageway and put through, each spill over the passageway and all extend to the bellied top surface of column.
According to some embodiments of the utility model, constant temperature equipment includes at least a pair of heat transfer return bend, each around the inner wall setting of cavity body fill the hot-fluid in the heat transfer return bend, the temperature of hot-fluid is 37 ℃ -40 ℃.
According to some embodiments of the utility model, each the import and the export of heat transfer return bend all stretch out from the lateral wall of cavity body, each the import, the export of heat transfer return bend all put through to a hot-fluid feeding mechanism, each dispose the circulating pump between the import, the export of heat transfer return bend and the hot-fluid feeding mechanism.
According to some embodiments of the utility model, the bottom outside of cavity body is equipped with the base, set up the screw thread on the base and connect the hole.
According to some embodiments of the present invention, a brain piece fixing device is provided in the observation slot.
According to some embodiments of the invention, the platform body is made of a transparent material.
Drawings
The present invention will be further explained with reference to the accompanying drawings:
fig. 1 is a schematic structural diagram of a platform main body according to an embodiment of the present invention;
fig. 2 is a schematic view of the internal structure of the platform main body according to the embodiment of the present invention.
Detailed Description
This section will describe in detail the embodiments of the present invention, preferred embodiments of the present invention are shown in the attached drawings, which are used to supplement the description of the text part of the specification with figures, so that one can intuitively and vividly understand each technical feature and the whole technical solution of the present invention, but they cannot be understood as the limitation of the protection scope of the present invention.
In the description of the present invention, it should be understood that the orientation or positional relationship indicated with respect to the orientation description, such as up, down, front, rear, left, right, etc., is based on the orientation or positional relationship shown in the drawings, and is only for convenience of description and simplification of description, and does not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.
In the description of the present invention, a plurality of means are one or more, a plurality of means are two or more, and the terms greater than, less than, exceeding, etc. are understood as not including the number, and the terms greater than, less than, within, etc. are understood as including the number. If the first and second are described for the purpose of distinguishing technical features, they are not to be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of the technical features indicated.
In the description of the present invention, unless there is an explicit limitation, the words such as setting, installation, connection, etc. should be understood in a broad sense, and those skilled in the art can reasonably determine the specific meanings of the above words in combination with the specific contents of the technical solution.
Referring to fig. 1 and 2, an imaging platform adapted to tissue transparentization and isolated brain slice according to an embodiment of the present invention includes a platform main body 1, the platform main body 1 includes a cylindrical hollow body 11 and an observation platform 12 fixed above the hollow body 11, the observation platform 12 includes a bottom plate and a side wall plate, the bottom plate is circular, and the middle part of the bottom plate is provided with an observation groove 121 for placing the isolated brain slice; a thermostatic device arranged inside the hollow body 11; the artificial cerebrospinal fluid circulation mechanism is mounted on the hollow body 11 and is in communication with the observation tank 121.
According to the utility model discloses imaging platform of adaptation tissue transparence and separation brain piece, including platform main part 1, constant temperature equipment and artifical cerebrospinal fluid circulation mechanism, wherein platform main part 1 includes cavity body 11 and observation platform 12, and the middle part of the bottom plate of observation platform 12 is equipped with the observation groove 121 that is used for placing separation brain piece, and artifical cerebrospinal fluid circulation mechanism is linked together with observation groove 121 for the circulation of artifical cerebrospinal fluid in observation groove 121 can be controlled, has guaranteed to observe the circulation balance of artifical cerebrospinal fluid in observation groove 121, ensures separation brain piece keep active, and then improves the imaging effect; simultaneously, constant temperature equipment sets up in cavity body 11, makes the temperature of platform main part 1 can be controlled and keep under normal physiological temperature condition, guarantees separation brain piece survival rate to the utmost, ensures the inside neuron vitality of separation brain piece, and the experiment accuracy obtains very big improvement.
In the embodiment of the present invention, a columnar protrusion 122 is disposed in the middle of the bottom plate, the observation trough 121 is located at the middle position of the columnar protrusion 122, meanwhile, the top surface of the columnar protrusion 122 is provided with a first inlet 1221 and a first outlet 1222, and a second inlet 1211 and a second outlet are disposed in the observation trough 121 from bottom to top. More specifically, the artificial cerebrospinal fluid circulation mechanism of this embodiment includes liquid inlet pipe 31 and liquid outlet pipe 32, first inlet 1221 and second inlet 1211, first liquid outlet 1222 and second liquid outlet are respectively through liquid inlet pipe 31, the switch-on of liquid outlet pipe 32, first inlet 1221 and first liquid outlet 1222 are respectively through hose connection to a peristaltic pump, both peristaltic pumps are connected to in an artificial cerebrospinal fluid storage device, so set up, the peristaltic pump is as the velocity of flow control part in the artificial cerebrospinal fluid circulation mechanism, can accurately control the circulation speed of artificial cerebrospinal fluid in observation groove 121, realize real-time adjustment, thereby guarantee the flow balance of artificial cerebrospinal fluid in observation groove 121, guarantee the activity of the separation of brain piece. Further, a pair of buckles 123 are arranged on the outer wall of the side wall plate to fix the hose respectively, and the two buckles 123 are symmetrically arranged on the outer wall of the side wall plate. In certain embodiments, the artificial cerebrospinal fluid circulation mechanism is integrally formed with the platform body and is made by 3D printing techniques with good replication, consistency, stability.
More specifically, a recovery groove 124 is formed between the columnar protrusion 122 and the side wall plate, the side surface of the columnar protrusion 122 is recessed inward to form at least one overflow channel 1223 communicated with the recovery groove 124, and each overflow channel 1223 extends to the top surface of the columnar protrusion 122. As shown in fig. 1, the side surface of the cylindrical protrusion 122 of the present embodiment is recessed inward to form two overflow channels 1223 communicated with the recovery groove 124, and an included angle between the two overflow channels 1223 is 180 °. The artificial cerebrospinal fluid is slowly injected by the peristaltic pump, is injected into the first liquid inlet 1221 through the liquid inlet pipe 31, enters the observation tank 121 through the second liquid inlet 1211 through the connecting pipe 123, overflows to the first liquid outlet 1222 through the connecting pipe 123 after the observation tank 121 is filled with the artificial cerebrospinal fluid and the liquid level line exceeds the second liquid outlet of the observation tank 121, and is pumped out through the peristaltic pump connected with the liquid outlet pipe 32, so that the cyclic updating of the artificial cerebrospinal fluid is realized; when the pumping speed of the peristaltic pump at the liquid outlet pipe 32 is less than the pushing speed at the liquid inlet pipe 31 and the liquid level of the artificial cerebrospinal fluid in the observation tank 121 is higher than the height of the observation tank 121, the overflowed artificial cerebrospinal fluid can slowly flow to the recovery tank 124 through the overflow channel 1223, so that after the experiment is completed, the experimenter can recover the part of the artificial cerebrospinal fluid from the recovery tank 124.
In some embodiments of the utility model, the constant temperature equipment includes at least a pair of heat transfer return bend around the inner wall setting of cavity body 11, pours the hot-fluid into each heat transfer return bend, and the temperature of hot-fluid is 37 ℃ -40 ℃, in order to more press close to human body temperature, preferably keeps the hot-fluid temperature at 37 ℃. Furthermore, an inlet and an outlet of each heat exchange elbow extend out of the side wall of the hollow body 11, the inlet and the outlet of each heat exchange elbow are communicated with a hot fluid supply device, and peristaltic pumps serving as circulating pumps are arranged between the inlet and the outlet of each heat exchange elbow and the hot fluid supply device; the hot fluid in the hot fluid supply device is double distilled water or an oily heat preservation medium.
In some embodiments of the present invention, a brain slice fixing device is disposed in the observation groove 121 to ensure that the isolated brain slice can still keep the position fixed in the flowing artificial cerebrospinal fluid; simultaneously, in order to guarantee the imaging platform's of this adaptation tissue transparence and separation brain piece position fixing in the experimentation, be equipped with base 111 in the bottom outside of cavity body 11, be provided with sample information identification area on base 111 and seted up the screw thread and connect hole 1111, the screw thread connects the setting up of hole 1111 to make it to pass through the fix with screw on shockproof bench to ensure holistic stability, guarantee the imaging quality.
In some embodiments of the present invention, the platform body 1 is made of a transparent material. In the specific embodiment, the observation groove 121 is made of colorless and transparent glass, and the parts of the observation platform 12 except the observation groove 121 and the hollow body 11 are made of colorless and transparent photosensitive resin; further, an orientation recognition mark 1212 is provided in the observation groove 121.
When the imaging platform for adapting to tissue transparentization and in-vitro brain slice is practically applied to in-vitro brain slice culture and real-time observation imaging, the method comprises the following steps: a. the peristaltic pump is connected through the heat exchange bent pipe, hot fluid is filled into the heat exchange bent pipe, and the temperature in the platform main body 1 is raised and kept constant at 37 ℃; b. connecting the liquid inlet pipe 31 and the first liquid inlet 1221 through a peristaltic pump, pouring the artificial cerebrospinal fluid which is subjected to supersaturation incubation by mixed oxygen (95% O2 and 5% CO2) gas into the observation tank 121, and recovering the overflowed artificial cerebrospinal fluid through the liquid outlet pipe 32 to keep the liquid level in the observation tank 121 stable; c. placing the in vitro brain slice sample on the bottom plate of the observation groove 121, and placing the brain slice fixing device on the in vitro brain slice sample; d. after incubation, the imaging platform of the adaptation tissue transparentization and in vitro brain slice is placed on the object carrying platforms of different imaging microscopes (platforms supporting laser confocal microscopes, multi-photon microscopes and the like); e. under the microscope, the view of the target area is quickly found through the position identification mark in the observation groove 121, and imaging is carried out.
When the imaging platform is used for tissue transparentization imaging, the following steps are required: a. placing the tissue sample after the transparentization treatment on a bottom plate of an observation tank 121, dripping a transparentization solution to immerse the tissue sample, and sealing the observation tank 121 by using a glass slide which is matched with the diameter of the bottom plate of the observation tank 121; b. placing imaging platforms of the adaptation tissue transparentization and in-vitro brain slices on object carrying platforms of different imaging microscopes (platforms supporting laser confocal microscopes, multi-photon microscopes and the like); c. under the microscope, the view of the target area is quickly found through the position identification mark in the observation groove 121, and imaging is carried out.
Naturally, the invention is not limited to the above-described embodiments, and those skilled in the art can make equivalent modifications or substitutions without departing from the spirit of the invention, and such equivalent modifications or substitutions are included in the scope of the claims of the present application.
Claims (10)
1. Imaging platform of adaptation tissue transparentization and separation brain piece, its characterized in that includes:
the platform main body comprises a hollow body and an observation platform fixed above the hollow body, wherein the observation platform comprises a bottom plate and a side wall plate, and the middle part of the bottom plate is provided with an observation groove for placing an in-vitro brain slice;
the constant temperature device is arranged in the hollow body;
and the artificial cerebrospinal fluid circulating mechanism is arranged in the hollow body and is communicated with the observation groove.
2. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 1, wherein: the middle part of bottom plate is provided with the column arch, it is located the bellied intermediate position of column to observe the trench, first inlet and first liquid outlet have been seted up to the bellied top surface of column, be equipped with second inlet and second liquid outlet from bottom to top in the observation trench.
3. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 2, wherein: the artificial cerebrospinal fluid circulating mechanism comprises a liquid inlet pipe and a liquid outlet pipe, and the first liquid inlet and the second liquid inlet and the first liquid outlet and the second liquid outlet are communicated through the liquid inlet pipe and the liquid outlet pipe respectively.
4. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 3, wherein: first inlet and first liquid outlet are connected to a peristaltic pump through the hose respectively, two the peristaltic pump all is connected to in an artificial cerebrospinal fluid storage device, the outer wall of side bounding wall is equipped with the buckle with the hose.
5. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 2, wherein: a recovery groove is formed between the columnar bulge and the side wall plate, the side surface of the columnar bulge is inwards sunken to form at least one overflow channel communicated with the recovery groove, and each overflow channel extends to the top surface of the columnar bulge.
6. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 1, wherein: the constant temperature device comprises at least one pair of heat exchange bent pipes arranged around the inner wall of the hollow body, hot fluid is filled in each heat exchange bent pipe, and the temperature of the hot fluid is 37-40 ℃.
7. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 6, wherein: the inlet and the outlet of each heat exchange elbow pipe extend out of the side wall of the hollow body, the inlet and the outlet of each heat exchange elbow pipe are communicated with a hot fluid supply device, and a circulating pump is arranged between the inlet and the outlet of each heat exchange elbow pipe and the hot fluid supply device.
8. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 1, wherein: the bottom outside of cavity body is equipped with the base, set up the screw thread and connect the hole on the base.
9. The imaging platform for adapting tissue clearing and ex vivo brain slices of claim 1, wherein: and a brain slice fixing device is arranged in the observation groove.
10. The imaging platform for adapting tissue clearing and ex vivo brain slices according to any one of claims 1 to 9, wherein: the platform main body is made of a transparent material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020241541.4U CN212008339U (en) | 2020-03-02 | 2020-03-02 | Imaging platform adaptive to tissue transparentization and in-vitro brain slice |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020241541.4U CN212008339U (en) | 2020-03-02 | 2020-03-02 | Imaging platform adaptive to tissue transparentization and in-vitro brain slice |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN212008339U true CN212008339U (en) | 2020-11-24 |
Family
ID=73425005
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202020241541.4U Active CN212008339U (en) | 2020-03-02 | 2020-03-02 | Imaging platform adaptive to tissue transparentization and in-vitro brain slice |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN212008339U (en) |
-
2020
- 2020-03-02 CN CN202020241541.4U patent/CN212008339U/en active Active
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP6893540B2 (en) | Gastric systems, devices and methods for use with enteral nutrition | |
| Shih et al. | A polished and reinforced thinned-skull window for long-term imaging of the mouse brain | |
| US20110229961A1 (en) | Active microfluidic system for in vitro culture | |
| CN206115320U (en) | Continuous perfusion system of isolated organ | |
| CN212008339U (en) | Imaging platform adaptive to tissue transparentization and in-vitro brain slice | |
| Belle et al. | Perforated multi-electrode array recording in hypothalamic brain slices | |
| KR101706153B1 (en) | Microfluidic chip-based cell cultivation system | |
| CN107197823A (en) | A kind of Establishment of Rat Model method of NASH | |
| Chan et al. | Passive driving forces of proximal tubular fluid and bicarbonate transport: gradient dependence of H+ secretion | |
| Yang et al. | A hybrid titanium-softmaterial, high-strength, transparent cranial window for transcranial injection and neuroimaging | |
| Qasimov | SPREAD OF DISEASES OF METABOLISM DISORDERS IN FISH | |
| CN205556644U (en) | Separation heart perfusion device suitable for rat | |
| JP5024777B2 (en) | Cell dynamics observation device | |
| CN203661818U (en) | Fish vision and temperature response research device | |
| CN205215424U (en) | Cornea marker suitable for porcine cornea transfer operation | |
| KR102429707B1 (en) | Automatic apparatus for supplying cell culture media with osmosis pump | |
| CN205607962U (en) | Eye excrement detector | |
| CN210198971U (en) | Portable body fluid observation device convenient to count | |
| Ma et al. | Imaging neuronal responses in slice preparations of vomeronasal organ expressing a genetically encoded calcium sensor | |
| CN202084236U (en) | Perfusion structure of mesentery microcirculation viewing device | |
| CN209383810U (en) | A kind of tissue incubating device | |
| CN215460772U (en) | Enema bag of accurate regulation and control | |
| CN203235139U (en) | Insulin skin testing template | |
| CN117660183B (en) | Embryo microfluidic dynamic culture dish and culture method thereof | |
| CN202134148U (en) | Insulation and moisture retention structure of mesentery microcirculation viewing device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant |