CN104287865B - A fixed observation device for live imaging of animal spinal cord - Google Patents

Abstract

A kind of animal spinal cord living imaging Orientation observation device, it by base plate (1), left side move adjusting means (2), adjusting means (3) is moved on right side and implanted observation window (4) forms；The mobile adjusting means in left side (2), right side are moved adjusting means (3) and are fixed on base plate (1) by screw, implanted observation window (4) by the rectangular opening on its V-type frame and left side move adjusting means (2), right side is moved the rectangular preiection of the horizontal location slide block in adjusting means (3) and is plugged together positioning.The present invention can either be conducive to the installation of implanted observation window to perform the operation, and the stability making backbone is more preferable, can be conducive to again the dynamic observation of postoperative two-photon spinal cord imaging.Left and right side moves the horizontal location slide block in adjusting means can not only provide horizontal direction stable interior stationary state to the operation of implanting device, gives stable extenal fixation and support when more can observe after surgery.

Description

A fixed observation device for live imaging of animal spinal cord

Technical field:

The invention relates to a fixed observation device for living animal spinal cord imaging, which is mostly used in experimental research in the field of neurobiology, can perform implanted observation window installation operations on animals, and can perform long-term dynamic observation of spinal cord nerve tissue and its components in vivo. Changes in blood vessels are of great value in the study of the central nervous system, especially the physiological and pathological changes of spinal cord injury, as well as the regeneration and repair of nerves after spinal cord injury.

Background technique:

In the experimental research in the field of neurobiology, the traditional method of in vivo imaging of the animal spinal cord is to excise a segment of the lamina to expose the spinal cord, use the whole body suspension method or local clamping to stabilize the spine, and then use the fluorescence microscope or two-photon Microscopy for in vivo dynamic imaging of exposed animal spinal cords. After the imaging, the muscles and skin were sutured layer by layer, and the skin and muscles were incised for the next observation to expose the original spinal cord segment and repeated. However, this process cannot last for a long time. Prolonged anesthesia and repeated operations will not only harm the animal's systemic function, but also increase the risk of infection, increase the inflammatory response, cause tissue damage, and cause more pain for the animal. Not conducive to further observation and experimental research. Moreover, there is currently no unified standard surgical fixation and postoperative observation external fixator, resulting in complicated surgical operations and difficulties in postoperative imaging and fixation.

Invention content:

1. Purpose: The purpose of the present invention is to provide a fixed observation device for living animal spinal cord imaging. It can not only facilitate the installation operation of the implanted observation window, improve the stability of the spine, but also facilitate the dynamic observation of postoperative two-photon spinal cord imaging. The horizontal positioning slider of the in vivo imaging external fixator can not only provide a stable internal fixation state in the horizontal direction for the operation of the implanter, but also provide stable external fixation support during postoperative observation.

2. Technical solution: The technical solution of the present invention is: a fixed observation device for live imaging of animal spinal cord, which is composed of a base plate 1, a left side movement adjustment device 2, a right side movement adjustment device 3 and an implanted observation window 4; The positional connection relationship between them is: the left moving adjustment device 2 and the right moving adjusting device 3 are fixed on the bottom plate 1 through screws, and the implanted observation window 4 moves with the left side through the rectangular hole on the V-shaped frame. The rectangular protrusions of the horizontal positioning slider on the adjustment device 2 and the right side movement adjustment device 3 are inserted and positioned.

The base plate 1 is a rectangular sheet material, and two mounting threaded holes are respectively provided on the left and right sides thereof for fixing the left moving adjusting device 2 and the right moving adjusting device 3 .

Described left side movement regulating device 2 is made up of dovetail groove base 2-1, horizontal positioning slide block 2-2 and positioning screw 2-3; The direction is dovetail-shaped, which can make the same dovetail-shaped horizontal positioning slider 2-2 reciprocate left and right in it; when sliding to the target position, tighten the positioning screw 2-3, and the positioning screw withstands the dovetail groove The base 2-1 achieves the purpose of preventing the relative sliding of the dovetail base 2-1 and the horizontal positioning slider 2-2. The shape of the dovetail groove base 2-1 is right-angled, with a dovetail-shaped groove in the horizontal direction and a rectangular block in the vertical direction. The horizontal positioning slider 2-2 is processed from a rectangular sheet material, and the screw holes provided on it are used for connection and positioning with the positioning screw 2-3, and the ends are processed into rectangular protrusions to facilitate matching with the V-shaped frame. Rectangular holes are inserted and positioned; the tail is processed into a dovetail form to facilitate sliding in the dovetail groove base 2-1. The positioning screws 2-3 are commercially available connectors.

The right side movement adjustment device 3 has the same structure as the left side movement adjustment device 2, and is installed mirror image symmetry;

The implantable observation window 4 is composed of a left V-shaped frame 4-1, a right V-shaped frame 4-2, a strip gasket 4-3, a rectangular cover glass 4-4 and mounting screws 4-5. composition; the positional connection relationship between them is: the left side V-shaped frame 4-1 and the right side V-shaped frame 4-2 are each provided with two threaded holes, after the position is fixed, the strip gasket 4-3 Place vertically on the threaded holes of the V-shaped frame on the left and right sides, and fix the strip gasket 4-3 to the V-shaped frame on the left and right sides by installing screws 4-5. In the middle of the two strip spacers 4-3, a rectangular cover glass 4-4 can be inserted. The distance between the left side V-shaped frame 4-1 and the right side V-shaped frame 4-2 can be regulated by the adjustment hole of the strip gasket 4-3.

Working principle of the present invention and flow process brief introduction are as follows:

The working principle of the present invention is to realize the purpose of installing the implanted observation window for the animal spinal cord through the external fixator for live imaging and the implanted observation window, and then use the two-photon microscope to perform living imaging of the spinal cord after the operation, thereby completing the living observation of the spinal cord Manipulation of dynamic changes in nerve cells.

Work process of the present invention is: for further illustrating technical scheme of the present invention, now the operation process that the present invention carries out is simply described as follows:

1. Implantable observation window installation operation: anesthetize the animal, shave the back and disinfect it with iodine, place it on the bottom plate, cut the skin at the projection of the chest 13-lumbar 2 in the center of the back of the animal, and clean the thorax 13-lumbar 2 vertebrae Muscles and soft tissues around the plate, revealing the thorax 13-lumbar 2 lamina.

2. Insert the horizontal positioning sliders on both sides into the slot holes of the V-shaped frame, that is, put the V-shaped frames on both sides into the horizontal positioning sliders, slide the horizontal positioning sliders horizontally, so that the V-shaped frames on both sides clamp the chest 13-waist 2 vertebrae.

3. Use micro scissors to resect the lumbar 1 lamina and fully expose the lumbar 1 spinal cord segment.

4. Place the strip-shaped spacers on both sides of the V-shaped frame and above the spinal cord, and tighten the four screws.

5. Cover the surface of the spinal cord with a layer of biopolysilicone resin rubber, and cover the resin with a rectangular cover glass. Use quick-drying superglue and tray powder to fix and seal the implanted observation window. After the superglue and tray powder are solidified, move the horizontal positioning sliders on both sides out of the slots of the V-shaped frame on both sides, and suture the skin to complete the operation.

6. Observation and fixation by two-photon microscope: Anesthetize the animal and place it in the center of the external fixation frame for live imaging. Slide the horizontal positioning sliders on both sides horizontally, extend into the slot holes on both sides of the V-shaped frame, and fix the implanted observation window and the animal spine.

7. In vivo neuroimaging of animal spinal cords using two-photon microscopy.

8. After the observation is completed, move the horizontal positioning sliders on both sides out of the slot holes of the V-shaped frame on both sides.

The present invention has the following advantages:

1. The implanted observation window is conducive to the long-term dynamic observation of the animal spinal cord using a two-photon microscope, avoiding the problems caused by repeated operations.

2. The external fixator is not only beneficial to the installation operation of the implanted observation window, but also more conducive to the fixation of the postoperative spinal cord observation of the animal.

3. The system has a wide range of applications. After changing the size, it can be used for large experimental animals such as rats, rabbits, dogs, and monkeys.

Description of drawings

Fig. 1 is a structural diagram of the fixed observation device for live imaging of the animal spinal cord.

Fig. 2a is a structural diagram of an implantable observation window.

Figure 2b is a schematic diagram of a V-shaped frame.

Fig. 2c is a schematic diagram of a strip-shaped gasket.

Figure 2d is a schematic diagram of a rectangular coverslip.

Fig. 3a is a structural diagram of the movement adjusting device.

Fig. 3b is a schematic diagram of the horizontal positioning slider.

Figure 3c is a schematic diagram of the dovetail base.

Figure 4 is a schematic diagram of the bottom plate.

The serial numbers in the figure are explained as follows:

1. Bottom plate; 2. Left mobile adjustment device; 3. Right mobile adjustment device; 4. Implantable observation window;

2-1. Dovetail base; 2-2. Horizontal positioning slider; 2-3. Positioning screw;

4-1, left V-shaped frame; 4-2, right V-shaped frame; 4-3, strip gasket; 4-4, rectangular cover glass; 4-5, mounting screws.

detailed description

See Fig. 1-Fig. 4, the technical scheme of the present invention is: a kind of fixed observation device of living imaging of animal spinal cord, it is made of base plate 1, left side movement adjustment device 2, right side movement adjustment device 3, implantable observation window 4 Composition; the positional connection relationship between them is: the left mobile adjustment device 2 and the right mobile adjustment device 3 are fixed on the bottom plate 1 through screws, and the implanted observation window 4 is connected to the left mobile adjustment device through the upper rectangular hole. 2. The rectangular protrusion of the right side movement adjustment device 3 is inserted and positioned.

The installation 1 is a rectangular plate, and two mounting threaded holes are respectively arranged on the left and right sides for fixing the left side movement adjustment device 2 and the right side movement adjustment device 3 .

Described left side movement regulating device 2 is made up of dovetail groove base 2-1, horizontal positioning slide block 2-2 and positioning screw 2-3; The direction is dovetail-shaped, which can make the same dovetail-shaped horizontal positioning slider 2-2 reciprocate left and right in it; when sliding to the target position, tighten the positioning screw 2-3, and the positioning screw withstands the dovetail groove The base 2-1 achieves the purpose of preventing the relative sliding of the dovetail base 2-1 and the horizontal positioning slider 2-2. The shape of the dovetail groove base 2-1 is right-angled, with a dovetail-shaped groove in the horizontal direction and a rectangular block in the vertical direction. The horizontal positioning slider 2-2 is processed from a rectangular sheet material, and the screw holes provided on it are used for connection and positioning with the positioning screw 2-3, and the ends are processed into rectangular protrusions to facilitate matching with the V-shaped frame. Rectangular holes are mated and positioned; the tail is processed into a dovetail form to facilitate sliding in the dovetail groove base 2-1. The positioning screws 2-3 are commercially available connectors.

The right side movement adjustment device 3 has the same structure as the left side movement adjustment device 2, and is installed in a mirror image;

The implantable experimental platform 4 is composed of a left V-shaped frame 4-1, a right V-shaped frame 4-2, a strip gasket 4-3, a rectangular cover glass 4-4 and mounting screws 4-5 composition; the position connection relationship between them is: there are two threaded holes on the left V-shaped frame 4-1 and the right V-shaped frame 4-2 respectively, after the position is fixed, the strip gasket 4-3 is vertically Place it on the threaded holes of the V-shaped frame on the left and right sides, and fix the strip-shaped gasket 4-3 to the V-shaped frame on the left and right sides through the mounting screws 4-5. In the middle of the two strip spacers 4-3, a rectangular cover glass 4-4 can be inserted. The distance between the left side V-shaped frame 4-1 and the right side V-shaped frame 4-2 can be regulated by the adjustment hole of the strip gasket 4-3.

The above description of the present invention with reference to the accompanying drawings is schematic and not restrictive. Those skilled in the art should be able to understand that in actual implementation, some changes may occur in the shape and layout of each component in the present invention; Under the enlightenment of the present invention, other persons may also make designs similar to the present invention or make modifications to the present invention and equivalent replacement of certain components. In particular, it should be pointed out that as long as they do not deviate from the design spirit of the present invention, all obvious changes and similar designs with equivalent replacements are included in the protection scope of the present invention.

Claims (1)

1. an animal spinal cord living imaging Orientation observation device, it is to be moved adjusting means (2), right side by base plate (1), left side Mobile adjusting means (3) and implanted observation window (4) composition；Adjusting means (3) is moved on the mobile adjusting means (2) in left side, right side Being fixed on base plate (1) by screw, implanted observation window (4) moves regulation dress by the rectangular opening on its V-type frame and left side Put (2), the rectangular preiection of horizontal location slide block on adjusting means is moved on right side (3) plugs together positioning；

It is characterized in that:

It is by dovetail groove base (2-1), horizontal location slide block (2-2) and positioning screw (2-that adjusting means (2) is moved in described left side 3) form；This dovetail groove base (2-1) is dovetail-shape in the horizontal direction so that the horizontal location being similarly swallow-tail form is sliding Block (2-2) bilateral reciprocation within it；When sliding into target location, screwing positioning screw (2-3), positioning screw withstands swallow Stern notch base (2-1), reaches to prevent the purpose of dovetail groove base (2-1) slip relative with horizontal location slide block (2-2)；This dovetail The rectangular shape of bottom land seat (2-1) profile, horizontal direction is provided with swallow tail shaped slot, and vertical direction is rectangular block；This horizontal location slide block (2-2) being that rectangular slab materials and parts process, the screw that it is arranged is for the connection and positioning, end with positioning screw (2-3) Being processed into rectangular preiection to be easy to plug together positioning with the rectangular opening on wishbone, afterbody is processed into dovetail form to be easy at the bottom of dovetail groove Slide in seat (2-1)；

Described base plate (1) is one piece of rectangle plate, and both sides are each provided with two installation screwed holes and move tune for left side around Regulating device (2), right side move the fixing of adjusting means (3)；

Adjusting means (3) and left side are moved in described right side, and to move adjusting means (2) structure identical, and specular is installed；

Described implanted observation window (4) is by left side wishbone (4-1), right side wishbone (4-2), strip pad (4-3), rectangular Shape cover glass (4-4) and installation screw (4-5) composition；It is respectively provided with two spiral shells on left side wishbone (4-1) and right side wishbone (4-2) Strip pad (4-3), after position is fixing, is vertically arranged on left and right sides wishbone screwed hole by pit, by installing screw (4- 5) strip pad (4-3) and left and right sides wishbone are fixed；In the middle of two strip pads (4-3), insert rectangle cover glass (4-4), the distance between left side wishbone (4-1) and right side wishbone (4-2) is then entered by the adjustment hole of strip pad (4-3) Row regulation.