A fixing device for repeated positioning imaging of animal back skin window

Abstract

The present invention belongs to the field of medical devices, and specifically relates to a fixing device for repeated positioning and imaging of a skin window on the back of an animal, comprising: a fixing frame, a fixing base, a translation stage for vertically adjusting the imaging focal length, a porous breadboard, and a two-dimensional translation stage; the fixing frame presents an arched structure, and its two end edges are detachably fixedly connected to the edges of the fixing base, and the central part of the fixing frame is provided with a fixing through hole that matches the position and diameter of the screw of the skin window on the back of the animal, and is used to fix the skin window on the back of the animal; the area of ​​the fixing base is larger than the area of ​​the fixing frame to support the body parts of the animal other than the back part to be observed; the fixing base is stacked and fixed on the upper end surface of the translation stage for vertically adjusting the imaging focal length; the translation stage for vertically adjusting the imaging focal length is fixed on the two-dimensional translation stage through the porous breadboard; the position of the site to be observed is determined by the position of the two-dimensional translation stage, and the present invention can ensure long-term, multi-day tracking imaging of the same animal position.

Description

Technical Field

[0001] The present invention belongs to the field of medical devices, and more specifically, relates to a fixing device for repeated positioning imaging of a skin window on the back of an animal. Background Art

[0002] In life science research, many life phenomena were revealed through in vitro tissue research in the early days. However, the separation of the research object from the living tissue made it inevitable that the results obtained would be different from the actual situation in the living body, or even completely opposite. Therefore, current scientific research tends to conduct in vivo research on living organisms to reflect the actual biological processes.

[0003] At present, when performing live imaging of animal back skin windows, only the part of the live animal that needs to be imaged is operated on and fixed with a back skin window device. During the imaging process, the live animal is freely placed under a microscope for observation, and there is no fixing device that matches the back skin window device well. In live imaging, the entire skin window will shake under the influence of the animal's own breathing, heartbeat or incomplete anesthesia, causing the entire skin window to shake under the drive of the mouse, resulting in image misalignment and distortion, and even the area of ​​interest may be completely moved out of the microscope imaging field of view. In addition, when it is necessary to better quantify the biological processes occurring in the body, it is necessary to repeatedly locate and image the same position multiple times, but due to the limitations of the microscopic imaging field of view, it is difficult to repeatedly locate and image the same position, especially in a homemade open-frame microscope. Summary of the invention

[0004] In view of the defects of the prior art and the need for improvement, the present invention provides a fixing device for repeated positioning and imaging of a skin window on the back of an animal. Its purpose is to provide a fixing device for repeated positioning and imaging of a skin window on the back of an animal and a method of using the same device, which can ensure long-term, multi-day tracking imaging of the same animal position.

[0005] To achieve the above-mentioned object, according to one aspect of the present invention, there is provided a fixing device for repeated positioning imaging of a skin window on the back of an animal, characterized in that it comprises: a fixing frame, a fixing base, a translation stage for vertically adjusting the imaging focal length, a multi-hole breadboard, and a two-dimensional translation stage;

[0006] The fixing frame presents an arched structure, and its two end edges are detachably fixedly connected to the edges of the fixing base. The central part of the fixing frame is provided with a fixing through hole that matches the position and diameter of the screw of the animal's back skin window, and is used to fix the animal's back skin window; the area of ​​the fixing base is larger than that of the fixing frame, so as to support the body parts of the animal other than the back part to be observed;

[0007] The fixed base is stacked and fixed on the upper end surface of the translation stage for vertically adjusting the imaging focal length; the translation stage for vertically adjusting the imaging focal length is fixed on the two-dimensional translation stage through the porous breadboard;

[0008] The position of the site to be observed is determined by the position of the two-dimensional translation stage, and the position of the imaging system used for imaging the back of the animal is fixed.

[0009] Furthermore, the two-dimensional translation stage is fixed on an optical anti-vibration platform.

[0010] Furthermore, two ends of the fixing frame are fixedly connected to the fixing base by bolts.

[0011] Furthermore, a heating pad is included, which is used to be arranged on the fixed base to maintain the body temperature of the animal.

[0012] Furthermore, the fixed base is of a concave structure, wherein the concave low-position flat plate is stacked and fixedly connected with the translation stage for vertically adjusting the imaging focal length, and the height of the fixed base is determined by the imaging distance.

[0013] Furthermore, a light-through hole is provided in the central portion of the fixing frame for placing the animal back skin window, and the animal back skin window is aligned with the light-through hole.

[0014] Furthermore, it also includes a spiral column that matches the size of the light-through hole on the fixing frame, and the spiral column is used to be embedded in the light-through hole from the back of the fixing base to prevent the animal's back from sagging.

[0015] Furthermore, the translation stage for vertically adjusting the imaging focal length is a Z-axis single-axis manual translation stage or a three-dimensional manual translation stage.

[0016] The present invention also provides a device for repeated positioning imaging of a skin window on the back of an animal, comprising: an imaging system, and a fixing device for repeated positioning imaging of a skin window on the back of an animal as described above;

[0017] Wherein, the objective lens of the imaging system is located at the upper part of the fixing frame in the fixing device.

[0018] The present invention also provides a method for repeated positioning imaging of a skin window on the back of an animal, comprising:

[0019] Fixing the skin window with the animal's back fixed on a fixing frame on a fixing device for repeated positioning imaging of the skin window on the animal's back as described above;

[0020] The positions of the translation stage and the two-dimensional translation stage in the fixing device for vertically adjusting the imaging focal length are adjusted, and combined with the imaging system, repeated positioning imaging of the skin window on the back of the animal is achieved.

[0021] In general, the above technical solutions conceived by the present invention can achieve the following beneficial effects:

[0022] (1) The fixing frame of the present invention has an arched structure, and its two end edges are detachably fixedly connected to the edges of the fixed base, thereby ensuring the stability of the imaging area during the imaging process and the accuracy of the experimental results; the fixed position of the skin window on the fixing frame is fixed, and the fixed base is installed on two stacked translation stages that can move in three dimensions in total. Therefore, the imaging area of ​​interest can be quickly located, and repeated precise positioning of the same position can be achieved, and long-term, multi-day, and multiple tracking imaging can be achieved without the need to keep the animal and the fixing frame under the microscope.

[0023] (2) The fixed base has a concave structure, in which a concave low-position flat plate is stacked and fixedly connected to the translation stage. The height of the fixed base is determined by the imaging distance. The design of the concave structure satisfies the requirement of ensuring the imaging distance when the imaging system cannot move.

[0024] (3) A light hole is provided at the center of the fixed frame for placing the skin window on the back of the animal. The skin window on the back of the animal is aligned with the light hole. The introduction of the light hole can prevent the part of the back of the animal to be observed from protruding from the skin window due to a mass, thereby affecting the observation effect. In addition, the device also includes a spiral column matching the size of the light hole. The spiral column is used to be embedded in the light hole from the back of the fixed base to prevent the back of the animal from sagging. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 A schematic diagram of the structure of a device for fixing a skin window on the back of an animal provided by an embodiment of the present invention;

[0026] Figure 2 A schematic diagram of the structure of a two-dimensional electric translation stage in a device for fixing a skin window on the back of an animal provided by an embodiment of the present invention;

[0027] Figure 3 A schematic diagram of the structure of a multi-hole breadboard in a device for fixing a skin window on the back of an animal provided by an embodiment of the present invention;

[0028] Figure 4 A schematic diagram of the structure of a manual three-dimensional translation stage in a device for fixing a skin window on the back of an animal provided by an embodiment of the present invention;

[0029] Figure 5 A schematic structural diagram of a fixing base in a fixing device for a leather window on the back of an animal provided by an embodiment of the present invention;

[0030] Figure 6 A schematic diagram of the structure of an imaging fixing frame in an animal back skin window fixing device provided by an embodiment of the present invention;

[0031] Figure 7This is a schematic structural diagram of another imaging fixing frame in the animal back skin window fixing device provided in an embodiment of the present invention.

[0032] In all drawings, the same reference numerals are used to denote the same elements or structures, wherein:

[0033] 1 is a large-range two-dimensional electric translation stage, 1-1 is a fixed screw hole array, 1-2 is a base screw hole array, 2 is a multi-hole breadboard, 2-1 is a screw hole array, 2-2 is an M6 threaded hole array, 3 is a manual three-dimensional translation stage, 3-1 is the first M6 screw hole, 4 is a fixed base, 4-1 is a first flat plate portion, 4-2 is a second flat plate portion, 4-3 is a vertical plate connecting portion, 4-11 is a second M6 screw hole, 4-21 is a first fixed through hole, 5 is a heating pad, 6 is an imaging fixed frame, 6-1 is a third flat plate portion, 6-2 is a fourth flat plate portion, 6-3 is a vertical plate portion, 6-11 is a light hole, 6-12 is a second fixed through hole, 6-21 is a third fixed through hole, and 7 is a spiral column. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0035] Embodiment 1

[0036] A fixing device for repeated positioning imaging of a skin window on the back of an animal comprises: a fixing frame, a fixing base, a translation stage for vertically adjusting the imaging focal length, a multi-hole breadboard, and a two-dimensional translation stage.

[0037] The fixing frame presents an arched structure, and its two end edges are detachably fixedly connected to the edges of the fixed base. The central part of the fixing frame is provided with a fixed through hole that matches the position and diameter of the screw of the animal's back skin window, which is used to fix the animal's back skin window; the area of ​​the fixed base is larger than the area of ​​the fixing frame to support the body parts of the animal other than the back part to be observed; the fixed base is stacked and fixed on the upper end surface of the translation stage for vertically adjusting the imaging focal length; the translation stage for vertically adjusting the imaging focal length is fixed on the two-dimensional translation stage through a porous breadboard; the position of the site to be observed is determined by the position of the two-dimensional translation stage, and the position of the imaging system for imaging the animal's back is fixed. The two ends of the fixing frame are fixedly connected to the fixed base by bolts. It also includes a heating pad, which is used to be set on the fixed base to maintain the body temperature of the animal.

[0038] Preferably, the fixed base is of a concave structure, wherein the concave low-position flat plate is stacked and fixedly connected with a translation stage for vertically adjusting the imaging focal length, and the height of the fixed base is determined by the imaging distance.

[0039] The design of the concave structure ensures the imaging distance when the imaging system cannot move.

[0040] Preferably, a light hole is provided in the central part of the fixing frame for placing the skin window on the back of the animal, and the skin window on the back of the animal is aligned with the light hole. The introduction of the light hole can prevent the part to be observed on the back of the animal from protruding from the skin window due to a lump, thereby affecting the observation effect.

[0041] Preferably, it also includes a spiral column whose size matches the light-through hole on the fixing frame, and the spiral column is used to be embedded in the light-through hole from the back of the fixing base to prevent the animal's back from sagging.

[0042] The preferred translation stage for vertically adjusting the imaging focal length is a Z-axis single-axis manual translation stage or a three-dimensional manual translation stage. The manual translation stage is convenient for adjusting the imaging focal length.

[0043] Embodiment 2

[0044] A device for repeated positioning and imaging of a skin window on the back of an animal, comprising: an imaging system, and a fixing device for repeated positioning and imaging of a skin window on the back of an animal as described above; wherein the objective lens of the imaging system is located at the upper part of a fixing frame in the fixing device.

[0045] The relevant technical solution is the same as that in Embodiment 1 and will not be described in detail here.

[0046] Embodiment 3

[0047] A method for repeated positioning imaging of a skin window on the back of an animal, comprising:

[0048] Fix the skin window on the back of the animal on a fixing frame on a fixing device for repeated positioning and imaging of the skin window on the back of the animal as described above; adjust the positions of the translation stage and the two-dimensional translation stage in the fixing device for vertically adjusting the imaging focal length, and combine with the imaging system to achieve repeated positioning and imaging of the skin window on the back of the animal.

[0049] Specifically, the two-dimensional electric translation stage can be fixed on the optical anti-vibration platform by screws; the multi-hole breadboard can be fixed on the two-dimensional electric translation stage by screws; the three-dimensional manual translation stage can be fixed on the breadboard by screws; the base of the fixture can be fixed on the three-dimensional translation stage by screws; the heating pad can be placed on the base of the fixture, the heating pad switch can be turned on, and the heating pad can be allowed to work normally, so that the body temperature of the small animal can be constant during the experiment; the small animal can be fixed with an animal fixing frame, and then the fixing frame can be fixed on the base of the fixture, and the small animal can be placed on the heating pad; according to the experimental requirements, the position of the two-dimensional electric translation stage can be adjusted, live imaging of a specific area can be performed, the position of the area of ​​interest can be recorded, and then the two-dimensional electric translation stage can be used to accurately repeat the positioning imaging.

[0050] In order to make the features and advantages of the present invention more understandable, the following is a description using mouse back skin window imaging as a specific example:

[0051] For example, Figures 1 to 6 As shown, the fixing device includes a large-range two-dimensional electric translation stage 1, a multi-hole breadboard 2, a manual three-dimensional translation stage 3, a fixed base 4, a heating pad 5, and an imaging fixing frame 6. The back skin window of the small animal can be reliably fixed to achieve stable, long-term, multi-day repeated positioning imaging.

[0052] The large-range two-dimensional electric translation stage 1 has a travel range of 114mm×76mm and a minimum step of 10nm, which can achieve precise repeated positioning. It is provided with a fixed screw hole array 1-1 (three rows and two columns of M4 screw holes with a length spacing of 260mm and a height spacing of 65mm) and a base screw hole array 1-2 (three pairs of M5 screw holes with a spacing of 50mm). The base screw holes are used to fix the electric translation stage on the optical anti-vibration platform, while the fixed screw hole array can be connected to other devices.

[0053] The porous breadboard 2 is a rectangular aluminum material, and a screw hole array 2-1 (three rows and two columns of M4 screw holes with a length spacing of 260mm and a height spacing of 65mm) is distributed on its upper edge, which corresponds to the screw holes of the two-dimensional electric translation stage. Considering the adjustability of the fixed position of the three-dimensional translation stage, an M6 threaded hole array 2-2 is added inside the breadboard, all of which are M6 threaded holes with a spacing of 25mm×25mm. The breadboard is an adapter, which is used to fix the breadboard on the two-dimensional electric translation stage and also to fix the three-dimensional translation stage 3 (the adjustment range in the three directions of xyz is 13mm) on the breadboard, forming a three-dimensional translation stage with repeatable positioning and adjustable height.

[0054] The fixed base 4 is an irregular three-dimensional aluminum material, including a first flat plate portion 4-1, a second flat plate portion 4-2 and a vertical plate connecting portion 4-3. The vertical plate connecting portion 4-3 connects the first flat plate portion 4-1 and the second flat plate portion 4-2. The first flat plate portion is provided with four second M6 screw holes 4-11 for fixing the imaging fixing frame 6, and the second flat plate portion is provided with two first fixing through holes 4-21 with a diameter of 6 mm and a spacing of 25 mm, which match the two first M6 screw holes 3-1 with a spacing of 25 mm on the three-dimensional translation stage 3;

[0055] The imaging fixing frame 6 is an arched structure, including a third flat plate portion 6-1, a fourth flat plate portion 6-2, and a vertical plate portion 6-3. Among them, the vertical plate portion 6-3 connects the third flat plate portion 6-1 and the fourth flat plate portion 6-2. The third flat plate portion 6-1 is provided with a light-through hole 6-11 and a second fixing through hole 6-12 that penetrates the third flat plate portion 6-1, and the edge portion of the fourth flat plate portion 6-2 is provided with four third fixing through holes 6-21 with a diameter of 6 mm that penetrate the fourth flat plate portion 6-1, which are equal to the number of the second M6 screw holes 4-11 of the fixing device base 4; the heating pad 5 can be flexibly configured according to the experimental conditions, and a heating pad with a constant temperature of 37 degrees Celsius is selected here.

[0056] When using the fixed imaging frame to fix the experimental animal, the electric two-dimensional translation stage, the multi-hole panel, the three-dimensional translation stage and the fixing device base can be assembled and placed under the microscope objective. Then, the animal skin window fixing device is matched with the second fixed through hole 6-12 to align the imaging window with the light hole 6-11, and then the fixed imaging frame 6 and the experimental animal are stably placed on the heating pad 5, and the animal skin window is fixed to the fixing device base 4 by M6 screws.

[0057] Example 1: Repeated localization imaging of mouse back skin window

[0058] Step 1: Depilate the mouse's back and then perform a dorsal skin window surgery;

[0059] Step 2: Fix the mouse skin window device on the imaging fixture 6 through the fixing through hole 6-12 on the imaging fixture 6, and at this time, the back skin window of the mouse is just aligned with the light hole 6-11 on the imaging fixture;

[0060] Step 3: Place a heating pad 5 on the fixed base 4 and set the temperature of the heating pad to 37 degrees Celsius;

[0061] Step 4: Place the mouse body on the heating pad 5, align the imaging fixture 6 with the second M6 screw hole 4-11 on the fixture base 4 through the fixing through hole 6-21, and fix the imaging fixture 6 on the fixture base 4 with screws;

[0062] Step 5: Use two-photon microscopy to find the region of interest, repeatedly observe and record the tumor microenvironment of the xenograft tumor in the mouse skin window for multiple days, and record the location of the region of interest;

[0063] Step 6: After imaging is completed, remove the anesthesia and keep the mouse on the heating pad until it wakes up. When the same mouse needs to be imaged later, repeated positioning imaging can be performed directly based on the previously recorded position.

[0064] See also Figure 7 , Figure 7 It is a structural schematic diagram of another fixing frame device for an animal back skin window in the present invention. In an embodiment of the present invention, the fixing device for the animal back skin window includes a large-range two-dimensional electric translation stage 1, a porous breadboard 2, a manual three-dimensional translation stage 3, a fixed base 4, a heating pad 5, and an imaging fixing frame 6.

[0065] The large-range two-dimensional electric translation stage 1 has a travel range of 114mm×76mm and a minimum step of 10nm, which can achieve accurate repeated positioning. It is provided with a fixed screw hole array 1-1 (three rows and two columns of M4 screw holes with a length spacing of 260mm and a height spacing of 65mm) and a base screw hole array 1-2 (three pairs of M5 screw holes with a spacing of 50mm). The base screw holes are used to fix the electric translation stage on the optical anti-vibration platform, while the fixed screw holes can be connected to other devices.

[0066] The porous breadboard 2 is a rectangular aluminum material, and a screw hole array 2-1 (three rows and two columns of M4 screw holes with a length spacing of 260mm and a height spacing of 65mm) is distributed on its upper edge, corresponding to the screw holes of the two-dimensional electric translation stage. Considering the adjustability of the fixed position of the three-dimensional translation stage, M6 threaded holes 2-2 are added inside the breadboard, all of which are M6 threaded holes with a spacing of 25mm×25mm. The breadboard is an adapter, which is used to fix the breadboard on the two-dimensional electric translation stage and also to fix the three-dimensional translation stage 3 (the adjustment range in the three directions of xyz is 13mm) on the breadboard, forming a three-dimensional translation stage with repeatable positioning and adjustable height as a whole;

[0067] The fixed base 4 is an irregular three-dimensional aluminum material, including a first flat plate portion 4-1, a second flat plate portion 4-2 and a vertical plate connecting portion 4-3. The vertical plate connecting portion 4-3 connects the first flat plate portion 4-1 and the second flat plate portion 4-2. The first flat plate portion is provided with four second M6 screw holes 4-11 for fixing the imaging fixing frame 6, and the second flat plate portion is provided with two first fixing through holes 4-21 with a diameter of 6 mm and a spacing of 25 mm, which match the two first M6 screw holes 3-1 with a spacing of 25 mm on the three-dimensional translation stage 3;

[0068] The imaging fixture 6 is an arched structure, including a third flat plate portion 6-1, a fourth flat plate portion 6-2, and a vertical plate portion 6-3. Among them, the vertical plate portion 6-3 connects the third flat plate portion 6-1 and the fourth flat plate portion 6-2. The third flat plate portion 6-1 is provided with a light-through hole 6-11 and a second fixed through hole 6-12 penetrating the third flat plate portion 6-1, and a thread 6-13 arranged on the inner wall of the light-through hole 6-11, combined with a spiral column 7 to prevent the sample from sinking during long-term imaging. The edge portion of the fourth flat plate portion 7-2 is provided with four third fixed through holes 6-21 with a diameter of 6 mm penetrating the fourth flat plate portion 7-1, which are equal to the number of the second M6 screw holes 4-11 of the fixing device base 4; the heating pad 5 can be flexibly configured according to the experimental conditions. Here, a heating pad with a constant temperature of 37 degrees Celsius is selected.

[0069] When using a fixed imaging frame to fix the experimental animal, the electric two-dimensional translation stage, the multi-hole breadboard, the three-dimensional translation stage and the fixed base can be assembled and placed under the microscope objective. Then, the animal skin window fixing device is matched with the second fixed through hole 6-12 to align the imaging window with the light hole 6-11, and then the fixed imaging frame 6 and the experimental animal are stably placed on the heating pad 5, and the animal skin window is fixed to the fixing device base 4 by M6 screws.

[0070] Example 2: Long-term imaging of the mouse back skin window;

[0071] Step 1: Depilate the mouse's back and then perform a dorsal skin window surgery;

[0072] Step 2: Fix the mouse skin window device on the imaging fixture 6 through the second fixing through hole 6-12 on the imaging fixture 6, at this time, the back skin window of the mouse is just aligned with the light hole 6-11 on the imaging fixture, and then align the spiral column 7 with the light hole 6-11 and screw it on to prevent the sample from sinking during the long-term imaging process;

[0073] Step 3: Place a heating pad 5 on the fixture base 4 and set the temperature of the heating pad to 37 degrees Celsius;

[0074] Step 4: Place the mouse body on the heating pad 5, align the imaging fixture 6 with the second M6 threaded hole 4-11 on the fixing base 4 through the third fixing through hole 6-21, and fix the imaging fixture 6 on the fixing base 4 with screws;

[0075] Step 5: Use two-photon microscopy to find the region of interest, perform repeated observations and records of the tumor microenvironment of the xenograft tumor in the mouse skin window over a long period of time for multiple days, and record the location of the region of interest;

[0076] Step 6: After imaging is completed, remove the anesthesia and keep the mouse on the heating pad until it wakes up. When the same mouse needs to be imaged later, repeated positioning imaging can be performed directly based on the previously recorded position.

[0077] The relevant technical solution is the same as that in Embodiment 1 and will not be described in detail here.

[0078] In summary, the fixing device for repeated positioning imaging of the back skin window of an animal proposed in the present invention can fix the back skin window of a living animal, ensure the stability of the position during the imaging process, and can accurately locate the imaging position, ensuring long-term, multi-day repeated positioning tracking imaging of the same position in the living imaging. The use of this device can reduce the error caused by jitter due to factors such as breathing and heartbeat during the experiment of living animals, and can repeatedly and accurately locate, which is beneficial to ensure the accuracy of the experimental results and improve the efficiency of data acquisition. Therefore, it can be mainly used for the fixation of small experimental animals such as mice, and for repeated positioning imaging of living animals for long periods of time for multiple days, so as to study the dynamic changes of various parameters in the microenvironment of the small experimental animals when they are awake or anesthetized.

[0079] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A fixing device for repeated positioning and imaging of a skin window on the back of an animal, characterized in that: It includes: a fixed frame, a fixed base, a translation stage for vertically adjusting the imaging focal length, a multi-hole breadboard, and a two-dimensional translation stage; The fixing frame presents an arched structure, and its two end edges are detachably fixedly connected to the edges of the fixing base. The central part of the fixing frame is provided with a fixing through hole that matches the position and diameter of the screw of the animal's back skin window, and is used to fix the animal's back skin window; the area of ​​the fixing base is larger than that of the fixing frame, so as to support the body parts of the animal other than the back part to be observed; The fixed base is stacked and fixed on the upper end surface of the translation stage for vertically adjusting the imaging focal length; the translation stage for vertically adjusting the imaging focal length is fixed on the two-dimensional translation stage through the porous breadboard; The position of the site to be observed is determined by the position of the two-dimensional translation stage, and the position of the imaging system used for imaging the back of the animal is fixed; A light-through hole is provided in the central part of the fixing frame for placing the animal back skin window, and the animal back skin window is aligned with the light-through hole; The fixing device also includes a spiral column that matches the size of the light-through hole on the fixing frame, and the spiral column is used to be embedded in the light-through hole from the back of the fixing base to prevent the animal's back from sagging; The imaging fixing frame is an arched structure, including a third flat plate portion, a fourth flat plate portion, and a vertical plate portion; wherein the vertical plate portion connects the third flat plate portion and the fourth flat plate portion; the third flat plate portion is provided with a light-through hole and a second fixing through hole penetrating the third flat plate portion, and the edge portion of the fourth flat plate portion is provided with four third fixing through holes penetrating the fourth flat plate portion, which is equal to the number of the second M6 screw holes of the base of the fixing device.

2. The fixing device according to claim 1, characterized in that: The two-dimensional translation stage is fixed on the optical anti-vibration platform.

3. The fixing device according to claim 1, characterized in that: The two ends of the fixing frame are fixedly connected to the fixing base by bolts.

4. The fixing device according to claim 1, characterized in that: Also included is a heating pad for being placed on the fixed base to maintain the body temperature of the animal.

5. The fixing device according to claim 1, characterized in that: The fixed base is of a concave structure, wherein the concave low-position flat plate is stacked and fixedly connected with the translation stage for vertically adjusting the imaging focal length, and the height of the fixed base is determined by the imaging distance.

6. The fixing device according to claim 1, characterized in that: The translation stage for vertically adjusting the imaging focal length is a Z-axis single-axis manual translation stage or a three-dimensional manual translation stage.

7. A device for repeated positioning imaging of a skin window on the back of an animal, characterized in that: It comprises: an imaging system, and a fixing device for repeated positioning imaging of a skin window on the back of an animal as described in any one of claims 1 to 6; Wherein, the objective lens of the imaging system is located at the upper part of the fixing frame in the fixing device.

8. A method for repeated positioning imaging of animal back skin windows, characterized in that: include: Fixing the skin window with the animal's back fixed on the fixing frame of the fixing device for repeated positioning imaging of the skin window on the animal's back as described in any one of claims 1 to 6; The positions of the translation stage and the two-dimensional translation stage in the fixing device for vertically adjusting the imaging focal length are adjusted, and combined with the imaging system, repeated positioning imaging of the skin window on the back of the animal is achieved.

Images