Small animal living imaging device and method

Abstract

The present invention discloses a small animal live imaging device and method. The small animal live imaging device includes: a stage with multiple light-transmitting windows for placing the small animal; a CMOS camera disposed above the stage; a brightfield light source disposed above the stage; a fluorescent light source assembly disposed above the stage; a filter wheel rotatably disposed below the CMOS camera, with multiple filters that transmit different wavelengths and at least one through-hole disposed on the same circumference of the filter wheel; a large-scale image sensor disposed below the stage and used for upward imaging; a vertical movement device disposed below the stage and used to drive the large-scale image sensor to move vertically; and a horizontal movement device disposed below the stage and used to drive the vertical movement device and the large-scale image sensor to move horizontally. In the present invention, fluorescent probe imaging and luminescent probe imaging of small animals can be achieved using the CMOS camera and the large-scale image sensor, respectively.

Description

Technical Field

[0001] The present invention belongs to the technical field of optical imaging, and in particular relates to a device and method for imaging small animal live bodies. Background Art

[0002] The CCD imaging solution collects the light signal emitted by the sample through a lens and transmits it to a CCD camera. Since the lens is at a certain distance from the sample being tested, the luminescence signal actually collected by the lens is only a small part of the light signal emitted by the sample detection side. In the in vivo imaging detection of small animals, the luminescence imaging signal from the small animals is very weak. Even a cooled CCD camera requires several minutes of exposure to capture the signal.

[0003] Domestic image sensor manufacturers mainly focus on CMOS chips, and some manufacturers also make sCOMS chips. No company is involved in the research and development and production of scientific-grade CCD chips. Therefore, in order to reduce costs and get rid of dependence on CCD cameras, in the field of small animal live imaging, it is necessary to design a device that can use CMOS cameras and sCOMS cameras for imaging. Summary of the Invention

[0004] The purpose of the present invention is to overcome the existing technical defects and provide a small animal living imaging device and method, which can respectively realize fluorescent probe imaging and luminescent probe imaging of small animals through a CMOS camera and a large-target image sensor.

[0005] In order to solve the above technical problems, the present invention provides a small animal living imaging device, comprising:

[0006] A loading platform, wherein the loading platform is provided with a plurality of light-transmitting windows for placing small animals;

[0007] A CMOS camera is provided above the stage and is used to photograph the small animals on the stage;

[0008] a bright field light source, disposed above the stage and used to illuminate the small animal on the stage;

[0009] a fluorescent light source assembly, disposed above the stage and used to illuminate the small animal on the stage;

[0010] a filter wheel rotatably disposed below the CMOS camera, wherein a plurality of filters transmitting different wavelengths and at least one through-hole are disposed on the same circumference of the filter wheel, so that different filters or through-holes are located directly below the CMOS camera during rotation of the filter wheel;

[0011] A large-target image sensor is provided below the stage and is used for upward shooting;

[0012] A vertical moving device, disposed below the stage and used to drive the large-target image sensor to move vertically;

[0013] The horizontal moving device is arranged below the object stage and is used to drive the vertical moving device and the large target image sensor to move horizontally.

[0014] Furthermore, the fluorescent light source assembly includes multiple fluorescent light sources with different wavelengths.

[0015] Furthermore, the fluorescent light sources are symmetrically arranged below the CMOS camera.

[0016] Furthermore, the bright field light source includes a plurality of monochromatic LED lamps.

[0017] Furthermore, five light-transmitting windows are arranged at intervals on the loading platform.

[0018] Furthermore, the light-transmitting window is made of transparent glass or acrylic plate.

[0019] Furthermore, a lens is provided at the lower end of the CMOS camera.

[0020] Furthermore, the small animal living imaging device also includes a background plate that is detachably arranged on the surface of the stage to shield the light-transmitting window.

[0021] In a second aspect, the present invention further provides a method for in vivo imaging of small animals, which uses the in vivo imaging device for small animals as described in any one of the first aspects to perform imaging, comprising the following steps:

[0022] S1. Place a background plate on the stage.

[0023] S2. Place the anesthetized animal with the fluorescent probe on the background plate;

[0024] S3, turning on the bright field light source, aligning the through-holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal;

[0025] S4, turning off the bright field light source, turning on the fluorescent light source corresponding to the fluorescent probe, and aligning the filter corresponding to the fluorescent probe on the filter wheel with the CMOS camera, and then capturing the image with the CMOS camera to obtain a fluorescent image;

[0026] S5. Overlaying the captured appearance image and the fluorescent image to obtain a superimposed image of the fluorescent signal and the appearance of the small animal, thereby obtaining the position of the fluorescent signal on the small animal.

[0027] In a third aspect, the present invention further provides another method for in vivo imaging of small animals, which uses the in vivo imaging device for small animals as described in any one of the first aspects to perform imaging, comprising the following steps:

[0028] S10, placing the anesthetized animals with luminous probes on the light-transmitting windows one by one;

[0029] S20, turning on the bright field light source, and aligning the through holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal at each position;

[0030] S30, turning off the brightfield light source, driving the vertical moving device and the horizontal moving device to position the large-scale image sensor closely below the first light-transmitting window, capturing a first luminescence signal image with the large-scale image sensor, and then moving the large-scale image sensor to below the second light-transmitting window and capturing a second luminescence signal image, until the large-scale image sensor is moved to the last light-transmitting window and capturing a final luminescence signal image;

[0031] S40, stitching together the multiple luminescence signal images obtained by sequentially capturing the images according to the movement distance of the large-scale image sensor to obtain a luminescence signal image with the same order and position as the actual small animals;

[0032] S50 , superimposing the photographed appearance image and the spliced ​​luminescence signal image to obtain a superimposed image of the luminescence signal at each position and the appearance of the small animal, thereby obtaining the position of the luminescence signal on the small animal.

[0033] In a fourth aspect, the present invention further provides another small animal in vivo imaging device, comprising:

[0034] A loading platform, wherein the loading platform is provided with a light-transmitting window for placing small animals;

[0035] A CMOS camera is provided above the stage and is used to photograph the small animals on the stage;

[0036] a bright field light source, disposed above the stage and used to illuminate the small animal on the stage;

[0037] a fluorescent light source assembly, disposed above the stage and used to illuminate the small animal on the stage;

[0038] a filter wheel rotatably disposed below the CMOS camera, wherein a plurality of filters transmitting different wavelengths and at least one through-hole are disposed on the same circumference of the filter wheel, so that different filters or through-holes are located directly below the CMOS camera during rotation of the filter wheel;

[0039] The large-target image sensor is closely arranged below the light-transmitting window and is used for shooting upward.

[0040] In a fifth aspect, the present invention further provides another method for in vivo imaging of small animals, which uses the in vivo imaging device for small animals as described in the fourth aspect to perform imaging, comprising the following steps:

[0041] S100, placing anesthetized small animals equipped with luminous probes on the light-transmitting window in an arranged manner;

[0042] S200, turning on the bright field light source, and aligning the through holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal at each position;

[0043] S300, turning off the bright field light source, and photographing with a large-area image sensor to obtain a luminescent signal image that is identical to the actual order and position of the small animals;

[0044] S400 , superimposing the captured appearance image and the luminous signal image to obtain an image of the luminous signal at each position superimposed with the appearance of the small animal, thereby obtaining the position of the luminous signal on the small animal.

[0045] The present invention has the following beneficial effects:

[0046] In the present invention, the combination of fluorescent light source and bright field light source with CMOS camera and large-target image sensor can be used for fluorescent probe imaging and luminescent probe imaging of small animals, wherein the fluorescent probe imaging signal is strong and the imaging exposure time is short, generally a few seconds, so the use of CMOS camera or sCMOS camera can meet the requirements of fluorescent probe imaging; and in view of the problem of long exposure time of CCD camera shooting, the exposure time can be shortened by collecting as many light signals as possible to reduce the requirements for imaging camera. In the present invention, during the luminescent probe imaging detection, a mobile device can be used to enlarge the target image sensor to shoot multiple small animals one by one, or a mobile device can be not used, and a larger target image sensor can be directly used to shoot multiple small animals as a whole, and the large target image sensor can be placed close to the bottom of the transparent window for shooting. Without using a lens, almost 100% of the light signals emitted by the detection side sample are collected, and the exposure time can also be completed in a few seconds. Therefore, the use of a large target image sensor can meet the requirements of luminescent probe imaging.

[0047] Additional aspects and advantages of the present invention will be set forth in part in the following description, will become apparent from the following description, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] The drawings described herein are used to provide a further understanding of the present invention, constitute a part of this application, and do not constitute an improper limitation of the present invention. In the drawings:

[0049] Figure 1 Schematic diagram of a small animal in vivo imaging device in an embodiment;

[0050] Figure 2 Schematic diagram of fluorescent probe imaging using a small animal in vivo imaging device in an embodiment;

[0051] Figure 3 Schematic diagram of the embodiment of the invention when the small animal in vivo imaging device is used to perform luminescent probe imaging;

[0052] Figure 4 Schematic diagram of the small animal in vivo imaging device in Example 4. DETAILED DESCRIPTION

[0053] In order to more fully understand the technical content of the present invention, the present invention will be further introduced and explained in conjunction with the accompanying drawings and specific embodiments below; it should be noted that if there are descriptions such as "first" and "second" in the text, they are used to distinguish different components, etc., and do not represent the order of precedence, nor do they limit the "first" and "second" to different types.

[0054] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making any creative work shall fall within the scope of protection of the present invention.

[0055] Example

[0056] like Figure 1 and Figure 2 As shown, the small animal live imaging device shown in this embodiment is installed in a chassis and specifically includes a stage 2, a CMOS camera 8, a brightfield light source 4, a fluorescent light source assembly 5, a filter wheel 7, a large target image sensor 11, a vertical movement device 12, and a horizontal movement device 13.

[0057] A plurality of light-transmitting windows 1 for placing the small animals 3 to be detected are arranged at intervals on the stage 2, so that the light source can illuminate the small animals 3 to be detected through the light-transmitting windows 1 for exposure and imaging.

[0058] The CMOS camera 8 is arranged above the stage for photographing the small animals 3 on the stage 2 downwards, and a lens 9 is provided at the lower end of the CMOS camera 8 , the lens angle 10 of which can cover all the small animals 3 on the stage 2 .

[0059] The bright field light source 4 is arranged above the stage 2 and is used to illuminate the small animals 3 on the stage 2. The bright field light source 4 can be installed on both sides of the top of the chassis or on both sides of the wall inside the chassis, so that the illumination range of the bright field light source 4 can cover all the small animals 3 on the stage 2, and is used for vertical downward illumination or inclined downward illumination.

[0060] The fluorescent light source assembly 5 is disposed above the stage 2 and is used to illuminate the small animal 3 on the stage 2 . The fluorescent light source assembly 5 is installed on the top of the chassis.

[0061] The filter wheel 7 is rotatably positioned below the CMOS camera 8. Several filters 6 transmitting different wavelengths and at least one through-hole (not shown) are provided on the same circumference of the filter wheel 7. As the filter wheel 7 rotates, different filters or through-holes on the same circumference are positioned directly below the lens 9 at the lower end of the CMOS camera 8. Consequently, different filters or through-holes can be used with the CMOS camera 8 according to different detection requirements.

[0062] The large-scale image sensor 11 is arranged below the stage 2 and is used to shoot upward, so that the large-scale image sensor 11 can be used to shoot the small animals 3 through the light-transmitting window 1; the vertical moving device 12 is arranged below the stage 2 and is used to drive the large-scale image sensor 11 to move vertically, so that the large-scale image sensor 11 can move toward or away from the stage 2, so as to adjust the field of view of the large-scale image sensor 11 when shooting during the vertical movement, and can change the amount of light entering the large-scale image sensor 11; the horizontal moving device 13 is arranged below the stage 2 and is used to drive the vertical moving device 12 and the large-scale image sensor 11 to move horizontally, that is, through horizontal movement, the large-scale image sensor 11 can move between each light-transmitting window 1 and shoot the small animals 3 located at the corresponding light-transmitting window 1 one by one.

[0063] In this embodiment, the fluorescent light source assembly 5 includes multiple fluorescent light sources of different wavelengths. Therefore, according to the actual detection requirements, several or more light sources with different wavelengths can be designed to respectively excite fluorescent probes of different colors. The multiple fluorescent light sources are symmetrically arranged on both sides below the CMOS camera 8. Of course, during installation, the fluorescent light sources need to be installed in a position that does not block the shooting angle of the lens.

[0064] In this embodiment, the fluorescent light source may be a luminous light source transmitted through an optical fiber, or may be at least one of an LED light source, an OLED light source, or a laser light source. The specific form of the fluorescent light source is not limited in this embodiment.

[0065] Preferably, the bright field light source 4 includes multiple monochromatic LED lights, such as green LEDs or red LEDs; preferably, in addition to being an LED light source, the bright field light source 4 can also be at least one of an OLED light source or a laser light source.

[0066] In this embodiment, five light-transmitting windows 1 are arranged at intervals on the stage 2, so that five small animals 3 can be imaged and detected in vivo at the same time, thereby improving the detection efficiency; preferably, the light-transmitting windows 1 are made of transparent glass or acrylic plate to facilitate light transmission.

[0067] In this embodiment, the small animal in vivo imaging device further includes a background plate 14 placed on the surface of the stage 2 , so that the background plate 14 shields the light-transmitting window 1 to facilitate fluorescence imaging detection.

[0068] In other embodiments, both the vertical moving device and the horizontal moving device may adopt a screw transmission mechanism, or a belt transmission mechanism or a cylinder transmission mechanism, etc. The screw transmission mechanism, the belt transmission mechanism and the cylinder transmission mechanism are all conventional transmission mechanisms.

[0069] Example 2

[0070] like Figure 2 As shown, a small animal in vivo imaging method shown in this embodiment uses the small animal in vivo imaging device as described in Example 1 to perform fluorescent probe imaging, including the following steps:

[0071] S1. Place a background plate 14 on the stage 2.

[0072] S2, placing multiple anesthetized small animals 3 with fluorescent probes on a background plate 14 in an orderly manner;

[0073] S3, turning on the bright field light source 4, and aligning the through hole on the filter wheel 7 with the lens 9 at the lower end of the CMOS camera 8, and then taking a picture through the through hole through the CMOS camera 8 and the lens 9 to obtain a bright field image, i.e., an image of the appearance of multiple small animals;

[0074] S4, then turning off the bright field light source 4, turning on the fluorescent light source corresponding to the color of the fluorescent probe, and aligning the filter 6 corresponding to the color of the fluorescent probe on the filter wheel 7 with the lens 9 at the lower end of the CMOS camera 8, so that the filter 6 filters out light sources other than the color of the fluorescent probe, and then capturing the light through the CMOS camera 8 and the lens 9 to obtain a fluorescent image, i.e., a fluorescent signal map;

[0075] S5. Since the same camera is used for shooting, the appearance image and the fluorescence image can be superimposed and overlapped to obtain a picture of the fluorescence signal and the appearance of the small animal, and then the position of the fluorescence signal on the small animal can be obtained. Quantitative analysis can then be performed based on the size and position of the fluorescence signal.

[0076] In the above description, during the entire imaging and detection process, an external anesthesia system needs to continuously supply anesthesia gas into the chassis to keep the small animal in an anesthesia state.

[0077] Example 3

[0078] like Figure 3 As shown, a small animal in vivo imaging method shown in this embodiment uses the small animal in vivo imaging device as described in Example 1 to perform luminescent probe imaging, including the following steps:

[0079] S10, at this time, no background plate is placed on the stage 2, and multiple small animals 3 with luminous probes and anesthetized are placed one by one on the light-transmitting windows 1, that is, one small animal is placed on each light-transmitting window 1;

[0080] S20, turning on the bright field light source 4, and aligning the through hole on the filter wheel 7 with the lens 9 at the lower end of the CMOS camera 8, and then taking a picture through the through hole through the CMOS camera 8 and the lens 9 to obtain a bright field image, i.e., an image of the appearance of multiple small animals at different positions;

[0081] S30, turning off the bright field light source 4, driving the vertical moving device 12 and the horizontal moving device 13 to move the large-target image sensor 11 closely to the bottom of the first light-transmitting window 1 arranged in a front-to-back manner, and taking pictures with the large-target image sensor 11. Since the luminescent probe can emit light by itself, a first luminescent signal image, i.e., a luminescent signal image of the first small animal 3, can be obtained. Then, the large-target image sensor 11 is moved to the bottom of the second light-transmitting window 1 and a second luminescent signal image, i.e., a luminescent signal image of the second small animal 3, is obtained by taking pictures. In this way, the large-target image sensor 11 is moved to the last light-transmitting window 1 and a last luminescent signal image, i.e., a luminescent signal image of the last small animal 3, is taken.

[0082] S40, stitching together the multiple luminescence signal images obtained by sequentially capturing the images according to the moving distance of the large-surface image sensor 11, that is, the distance between two adjacent luminescence signal images is the same as the distance between two adjacent small animals, thereby obtaining a combined luminescence signal image with the same order and position as the actual small animals;

[0083] S50. Superimpose and overlap the appearance image obtained by shooting and the combined luminous signal image obtained after splicing and combining, and obtain a picture of the luminous signal at each position and the appearance of the small animal superimposed, and then obtain the position of the luminous signal on the small animal, and then perform quantitative analysis based on the size and position of the luminous signal; here, although the appearance image and the combined luminous signal image are shot by different cameras and generally cannot be directly superimposed, by accurately positioning the distance of the horizontal moving device each time it shoots, a combined luminous signal image with the same order and position as the actual small animal can be obtained by splicing, so that the images shot by different cameras can also be superimposed up and down, thereby solving the problem of difficulty in superimposing between the appearance image and the combined luminous signal image.

[0084] In the above description, during the entire imaging and detection process, an external anesthesia system needs to continuously supply anesthesia gas into the chassis to keep the small animal in an anesthesia state.

[0085] Example 4

[0086] like Figure 4 As shown, a small animal living imaging device shown in this embodiment includes the small animal living imaging device installed in a chassis, specifically including a stage 2, a CMOS camera 8, a bright field light source 4, a fluorescent light source assembly 5, a filter wheel 7 and a large target area image sensor 11.

[0087] Among them, a light-transmitting window 1 for placing the small animal 3 to be detected is provided on the stage 2, so that the light source can be irradiated on the small animal 3 to be detected through the light-transmitting window 1 for exposure imaging; it can be understood that the light-transmitting window 1 can be a large window for placing multiple small animals, or a small window that can only place a single small animal. When a small window is used, a plurality of light-transmitting windows 1 arranged at intervals can be set on the stage 2 to place multiple small animals and then perform simultaneous detection.

[0088] The CMOS camera 8 is arranged above the stage for photographing the small animals 3 on the stage 2 downwards, and a lens 9 is provided at the lower end of the CMOS camera 8 , the lens angle 10 of which can cover all the small animals 3 on the stage 2 .

[0089] The bright field light source 4 is arranged above the stage 2 and is used to illuminate the small animals 3 on the stage 2. The bright field light source 4 can be installed on both sides of the top of the chassis or on both sides of the wall inside the chassis, so that the illumination range of the bright field light source 4 can cover all the small animals 3 on the stage 2, and is used for vertical downward illumination or inclined downward illumination.

[0090] The fluorescent light source assembly 5 is disposed above the stage 2 and is used to illuminate the small animal 3 on the stage 2 . The fluorescent light source assembly 5 is installed on the top of the chassis.

[0091] The filter wheel 7 is rotatably positioned below the CMOS camera 8. Several filters 6 transmitting different wavelengths and at least one through-hole (not shown) are provided on the same circumference of the filter wheel 7. As the filter wheel 7 rotates, different filters or through-holes on the same circumference are positioned directly below the lens 9 at the lower end of the CMOS camera 8. Consequently, different filters or through-holes can be used with the CMOS camera 8 according to different detection requirements.

[0092] The large-target image sensor 11 is closely arranged below the light-transmitting window 2 and is used to shoot upward, so that the small animal 3 can be photographed through the light-transmitting window 1 using the large-target image sensor 11; the shooting angle of the large-target image sensor 11 needs to cover the entire light-transmitting window, so as to simultaneously photograph the small animal placed on the light-transmitting window 1.

[0093] The method for performing fluorescent probe imaging detection using the small animal in vivo imaging device described in this embodiment is as described in Example 2.

[0094] The small animal in vivo imaging device described in this embodiment includes the following steps when performing luminescent probe imaging:

[0095] S100, placing multiple anesthetized small animals equipped with luminous probes on a light-transmitting window in an arranged manner at intervals;

[0096] S200, turning on the bright field light source, and aligning the through holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal at each position;

[0097] S300, turning off the bright field light source, and photographing with a large-area image sensor to obtain a luminescent signal image that is identical to the actual order and position of the small animals;

[0098] S400, superimpose the captured appearance image and the luminescence signal image to obtain a superimposed image of the luminescence signal at each position and the appearance of the small animal, and then obtain the position of the luminescence signal on the small animal, and then perform quantitative analysis based on the size and position of the luminescence signal.

[0099] The technical solutions provided by the embodiments of the present invention are introduced in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the embodiments of the present invention. The description of the above embodiments is only applicable to help understand the principles of the embodiments of the present invention. At the same time, for those skilled in the art, according to the embodiments of the present invention, there may be changes in the specific implementation methods and application scopes. In summary, the contents of this specification should not be understood as limiting the present invention.

Claims

1. A small animal living imaging device, characterized in that: include: A loading platform, wherein the loading platform is provided with a plurality of light-transmitting windows for placing small animals; A CMOS camera is provided above the stage and is used to photograph the small animals on the stage; a bright field light source, disposed above the stage and used to illuminate the small animal on the stage; a fluorescent light source assembly, disposed above the stage and used to illuminate the small animal on the stage; a filter wheel rotatably disposed below the CMOS camera, wherein a plurality of filters transmitting different wavelengths and at least one through-hole are disposed on the same circumference of the filter wheel, so that different filters or through-holes are located directly below the CMOS camera during rotation of the filter wheel; A large-target image sensor is provided below the stage and is used for upward shooting; A vertical moving device, disposed below the stage and used to drive the large-target image sensor to move vertically; The horizontal moving device is arranged below the object stage and is used to drive the vertical moving device and the large target image sensor to move horizontally.

2. The small animal living body imaging device according to claim 1, characterized in that: The fluorescent light source assembly includes a plurality of fluorescent light sources with different wavelengths, and the fluorescent light sources are symmetrically arranged below the CMOS camera.

3. The small animal living body imaging device according to claim 1, characterized in that: The bright field light source includes a plurality of monochromatic LED lamps.

4. The small animal living body imaging device according to claim 1, characterized in that: Five light-transmitting windows are arranged at intervals on the object carrier, and the light-transmitting windows are made of transparent glass or acrylic plates.

5. The small animal living imaging device according to any one of claims 1 to 4, characterized in that: A lens is provided at the lower end of the CMOS camera.

6. The small animal living body imaging device according to claim 5, characterized in that: It also includes a background plate that is detachably arranged on the surface of the stage to shield the light-transmitting window.

7. A method for in vivo imaging of small animals, using the in vivo imaging device for small animals according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1. Place a background plate on the stage. S2. Place the anesthetized animal with the fluorescent probe on the background plate; S3, turning on the bright field light source, aligning the through-holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal; S4, turning off the bright field light source, turning on the fluorescent light source corresponding to the fluorescent probe, and aligning the filter corresponding to the fluorescent probe on the filter wheel with the CMOS camera, and then capturing the image with the CMOS camera to obtain a fluorescent image; S5. Overlaying the captured appearance image and the fluorescent image to obtain a superimposed image of the fluorescent signal and the appearance of the small animal, thereby obtaining the position of the fluorescent signal on the small animal.

8. A method for in vivo imaging of small animals, using the in vivo imaging device for small animals according to any one of claims 1 to 6, characterized in that: The following steps are involved: S10, placing the anesthetized animals with luminous probes on the light-transmitting windows one by one; S20, turning on the bright field light source, and aligning the through holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal at each position; S30, turning off the brightfield light source, driving the vertical moving device and the horizontal moving device to position the large-scale image sensor closely below the first light-transmitting window, capturing a first luminescence signal image with the large-scale image sensor, and then moving the large-scale image sensor to below the second light-transmitting window and capturing a second luminescence signal image, until the large-scale image sensor is moved to the last light-transmitting window and capturing a final luminescence signal image; S40, stitching together the multiple luminescence signal images obtained by sequentially capturing the images according to the movement distance of the large-scale image sensor to obtain a luminescence signal image with the same order and position as the actual small animals; S50 , superimposing the photographed appearance image and the spliced ​​luminescence signal image to obtain a superimposed image of the luminescence signal at each position and the appearance of the small animal, thereby obtaining the position of the luminescence signal on the small animal.

9. A small animal living imaging device, characterized in that: include: A loading platform, wherein the loading platform is provided with a light-transmitting window for placing small animals; A CMOS camera is provided above the stage and is used to photograph the small animals on the stage; a bright field light source, disposed above the stage and used to illuminate the small animal on the stage; a fluorescent light source assembly, disposed above the stage and used to illuminate the small animal on the stage; a filter wheel rotatably disposed below the CMOS camera, wherein a plurality of filters transmitting different wavelengths and at least one through-hole are disposed on the same circumference of the filter wheel, so that different filters or through-holes are located directly below the CMOS camera during rotation of the filter wheel; The large-target image sensor is closely arranged below the light-transmitting window and is used for shooting upward.

10. A method for small animal in vivo imaging, using the small animal in vivo imaging device according to claim 9 for imaging, characterized in that: The following steps are involved: S100, placing anesthetized small animals equipped with luminous probes on the light-transmitting window in an arranged manner; S200, turning on the bright field light source, and aligning the through holes on the filter wheel with the CMOS camera, and then taking a picture with the CMOS camera to obtain an image of the appearance of the small animal at each position; S300, turning off the bright field light source, and photographing with a large-area image sensor to obtain a luminescent signal image that is identical to the actual order and position of the small animals; S400 , superimposing the captured appearance image and the luminous signal image to obtain an image of the luminous signal at each position superimposed with the appearance of the small animal, thereby obtaining the position of the luminous signal on the small animal.

Images