Animal fluorescence hyperspectral imager

Abstract

The present application relates to the technical field of optical imaging, in particular to an animal fluorescence hyperspectral imager. The animal fluorescence hyperspectral imager provided by the present application comprises a hyperspectral video imaging system, a light source lamp array system and a three-dimensional displacement system. The animal fluorescence hyperspectral imager can divide the image of a detected object into imaging rays and present the spectral information of the imaging rays on a two-dimensional CMOS, thereby realizing hyperspectral video imaging and greatly improving the resolution and accuracy of the image. The three-dimensional displacement system can realize precise movement and adjustment of a target sample in three-dimensional directions, thereby obtaining better imaging effect.

Description

Technical Field

[0001] The present invention relates to the technical field of optical imaging, in particular to an animal fluorescence hyperspectral imager. Background Art

[0002] In recent years, with the continuous advancement of life science research, the demand for high-resolution imaging of biological tissues and cellular structures has become increasingly urgent. Animals are important model organisms in biomolecular and cellular studies, and are widely used due to their similar biological and physiological characteristics. In the field of animal fluorescence imaging research, it is necessary to acquire the fluorescence signal of the sample through imaging technology to understand the sample's molecular structure and function.

[0003] While conventional imaging devices such as microscopes or monochromators can achieve fluorescence imaging, they suffer from slow imaging speeds and high sample requirements. Against this backdrop, animal fluorescence imaging based on CMOS image sensors has been developed, enabling high-speed imaging and rapid capture of fluorescence signals within animals. However, these devices have limitations during the imaging process, including slow imaging speeds, high sample requirements, and room for improvement in image quality.

[0004] To address these issues, several new animal fluorescence imagers have emerged in recent years. Among them, those based on hyperspectral imaging technology offer excellent spectral resolution and contrast, enabling the acquisition of spectral information from samples, leading to a better understanding of their structure and function. However, the imaging quality of existing animal fluorescence hyperspectral imaging systems remains to be improved, limiting research and application in the field. Summary of the Invention

[0005] The purpose of the present invention is to overcome the defects of the existing technology and to provide an animal fluorescence hyperspectral imager that can achieve high-quality animal fluorescence imaging and hyperspectral imaging.

[0006] To achieve the above objectives, the present invention adopts the following specific technical solutions:

[0007] The animal fluorescence hyperspectral imager provided by the present invention includes a hyperspectral video imaging system, a light source array system and a three-dimensional displacement system. The hyperspectral video imaging system includes a light source intensity controller, a laser light source, an optical fiber, a data acquisition card, a galvanometer controller, a high-speed CMOS, a first imaging zoom lens, an imaging reflector, a sleeve grating, a slit reflector, a slit sleeve, a scanning galvanometer, a spectrometer cage cube, a sleeve, a turning lens barrel, a filter module, a support cage rod and a scanning galvanometer cage cube.

[0008] The light source lamp array system includes a light source bracket, a light source platform, a sapphire glass window, a light source lamp array, and a laser. The light source bracket is used to support and fix the light source platform and the optical platform. The light source platform is fixed above the optical platform. The sapphire glass window is set on the light source platform. The light source lamp array is fixed on the lower surface of the optical platform, and the setting position of the light source lamp array corresponds to the setting position of the sapphire glass window. The laser is set on the light source lamp array and is used to receive the laser light emitted by the laser light source and irradiate the laser light to the target sample surface after passing through the sapphire glass window.

[0009] The laser light source is emitted by a laser source and then transmitted to the laser through an optical fiber after being adjusted by a light source intensity controller. The laser passes through a sapphire glass window and irradiates the surface of the target sample. The reflected light from the target sample passes through the sapphire glass window again and enters the turning lens barrel. The reflected light changes direction after passing through the turning lens barrel and enters the spectrometer cage cube through a sleeve.

[0010] The scanning galvanometer is disposed within a scanning galvanometer cage cube. The galvanometer controller is used to control the rotation of the scanning galvanometer. A visible light grating is disposed within the sleeve grating, and the direction of the visible light grating is perpendicular to the direction of the slit. The reflected light emitted from the spectroscopic cage cube passes through the filter module and the support cage rods in sequence before entering the scanning galvanometer cage cube. The reflected light is then reflected by the scanning galvanometer, passes through the slit sleeve and the slit reflector in sequence, and is incident on the sleeve grating to form a diffraction spectrum. The diffraction spectrum passes through the imaging reflector and is focused by the first imaging zoom lens onto the two-dimensional plane of the high-speed CMOS sensor. The high-speed CMOS stores the acquired spectral image on the data acquisition card.

[0011] The three-dimensional displacement system includes a stage, a loading plate, an X-axis scanning guide, an X-axis scanning motor, a Y-axis scanning guide, a Y-axis scanning motor, a Z-axis scanning platform, a Z-axis scanning motor, a Z-axis scanning guide and a Z-axis scanning lead screw. The stage is used to place the imaging target. The stage is fixedly set on the loading plate, which is installed on the X-axis scanning guide. The X-axis scanning motor is used to drive the loading plate to move along the X-axis direction on the X-axis scanning guide; the X-axis scanning guide is installed on the Y-axis scanning guide, and the Y-axis scanning motor is used to drive the X-axis scanning guide and the loading plate to move along the Y-axis direction on the Y-axis scanning guide; the Y-axis scanning guide is fixedly mounted on the Z-axis scanning platform, the Z-axis scanning platform is installed on the Z-axis scanning guide, the Z-axis scanning platform is connected to the Z-axis scanning lead screw, and the Z-axis scanning motor is used to drive the Z-axis scanning lead screw to drive the Z-axis scanning platform to move along the Z-axis direction on the Z-axis scanning guide.

[0012] Preferably, the animal fluorescence hyperspectral imager also includes an imaging CMOS and a second imaging zoom lens. A spectrometer is also provided in the spectroscopic cage cube for dividing the reflected light into two light paths, forward and upward. The forward reflected light enters the scanning galvanometer cage cube after passing through the filter module and the support cage rod, and the upward reflected light passes through the second imaging zoom lens and enters the imaging CMOS, and is imaged on the two-dimensional plane of the imaging CMOS sensor.

[0013] Preferably, there is at least one laser light source, and the number of light source intensity controllers is the same as the number of laser light sources, which are used to individually control each laser light source to meet the excitation requirements of different fluorescent materials at different wavelengths.

[0014] Preferably, the focal length of the second imaging zoom lens is 18-35 nm.

[0015] Preferably, the three-dimensional displacement system further includes a Z-axis scanning platform leveling knob, and the Z-axis scanning platform leveling knob is used to perform balance calibration on the Z-axis scanning platform.

[0016] The present invention can achieve the following technical effects:

[0017] 1. The animal fluorescence hyperspectral imager provided by the present invention can split the image of the detected object into imaging rays and present its spectral information on a two-dimensional CMOS, thereby realizing hyperspectral video imaging and greatly improving the resolution and accuracy of the image;

[0018] 2. The present invention uses a single light source of different wavelengths to excite the fluorescence on the target, and then filters out the noise to obtain clear fluorescence imaging results, which can be applied to fields such as medicine and biology;

[0019] 3. The animal fluorescence hyperspectral imager of the present invention is equipped with a three-dimensional displacement system, which can achieve precise movement and adjustment of the target sample in three dimensions, thereby obtaining better imaging effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Schematic diagram of the overall workflow of the animal fluorescence hyperspectral imager provided according to an embodiment of the present invention.

[0021] Figure 2 3 is a schematic structural diagram of an animal fluorescence hyperspectral imager provided according to an embodiment of the present invention.

[0022] Figure 3 3 is a schematic structural diagram of a hyperspectral video imaging system provided according to an embodiment of the present invention.

[0023] Figure 4 2 is a schematic structural diagram of a light source lamp array system provided according to an embodiment of the present invention.

[0024] Figure 5 3D is a schematic structural diagram of a three-dimensional displacement system provided according to an embodiment of the present invention.

[0025] Reference numerals include:

[0026] Animal fluorescence hyperspectral imager 1, hyperspectral video imaging system 2, light source array system 3, three-dimensional displacement system 4, data cable 5, computer host 6, display 7, light source intensity controller 201, laser light source 202, optical fiber 203, data acquisition card 204, optical platform 205, galvanometer controller 206, high-speed CMOS 207, first imaging zoom lens 208, imaging reflector 209, sleeve grating 210, slit reflector 211, slit sleeve 212, scanning galvanometer 213, imaging CMOS 214, second imaging zoom lens 215, spectroscopic cage Cube 216, sleeve 217, turning lens barrel 218, filter module 219, support cage rod 220, scanning galvanometer cage cube 221, light source bracket 301, light source platform 302, sapphire glass window 303, light source array 304, laser 305, imaging target 401, stage 402, stage 403, X-axis scanning guide rail 404, Y-axis scanning motor 405, Y-axis scanning guide rail 406, Z-axis scanning platform 407, Z-axis scanning platform leveling knob 408, Z-axis scanning motor 409, Z-axis scanning screw 410, and Z-axis scanning guide rail 411. DETAILED DESCRIPTION

[0027] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In the following description, identical modules are denoted by identical reference numerals. In the case of identical reference numerals, their names and functions are also identical. Therefore, their detailed description will not be repeated.

[0028] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and do not constitute a limitation of the present invention.

[0029] The embodiment of the present invention provides an animal fluorescence hyperspectral imager, Figure 1 The overall workflow of the animal fluorescence hyperspectral imager is shown. The animal fluorescence hyperspectral imager 1 collects hyperspectral or fluorescence imaging data of the imaging target and transmits the data to the computer host 6 in real time through the data line 5. After the computer host 6 analyzes and processes the data, the hyperspectral or fluorescence spectrum is displayed in real time through the display 7 for the user to view.

[0030] Figure 2 The structure of the animal fluorescence hyperspectral imager provided by the embodiment of the present invention is shown in FIG. Figure 2As shown, the animal fluorescence hyperspectral imager 1 includes a hyperspectral video imaging system 2, a light source array system 3 and a three-dimensional displacement system 4 from top to bottom. The hyperspectral video imaging system 2 is used for fluorescence hyperspectral imaging, the light source array system 3 is used to irradiate the laser on the imaging target to excite the corresponding fluorescence, and the three-dimensional displacement system 4 is used to adjust the presentation target position. Through the coordinated cooperation of the hyperspectral video imaging system 2, the light source array system 3 and the three-dimensional displacement system 4, high-speed, efficient and high-precision hyperspectral video imaging can be achieved.

[0031] Figure 3 The structure of the hyperspectral video imaging system provided by the embodiment of the present invention is shown in FIG. Figure 3 As shown, the hyperspectral video imaging system 2 includes a light source intensity controller 201, a laser light source 202, an optical fiber 203, a data acquisition card 204, a galvanometer controller 206, a high-speed CMOS 207, a first imaging zoom lens 208, an imaging reflector 209, a sleeve grating 210, a slit reflector 211, a slit sleeve 212, a scanning galvanometer 213, an imaging CMOS 214, a second imaging zoom lens 215, a spectroscopic cage cube 216, a sleeve 217, a turning lens barrel 218, a filter module 219, support cage rods 220, and a scanning galvanometer cage cube 221, which are installed on an optical platform 205.

[0032] The laser light source 202 excites a light source of a specific wavelength band. After the light source intensity is adjusted by the light source intensity controller 201, the light is irradiated onto the imaging target through the optical fiber 203. The light reflected from the imaging target passes through the deflection lens barrel 218 and the sleeve 217 and enters the spectrometer cube 216. The spectrometer cube 216 is equipped with a spectrometer to split the reflected light path into two paths: forward and upward. The upward light passes through the second imaging zoom lens 215 and is imaged on the two-dimensional plane of the imaging CMOS sensor 214. The forward light beam passes through the optical kit to achieve spectral imaging.

[0033] In a specific embodiment, the focal length range of the second imaging zoom lens 215 is 18 nm to 35 nm, which can conveniently achieve focusing effects of targets at different distances and different field sizes.

[0034] In a preferred embodiment, there is at least one laser light source 202, and the number of light source intensity controllers 201 is consistent with the number of laser light sources 202. Each laser light source 202 can be independently controlled by the light source intensity controller 201, which can realize the independent operation of different laser light sources 202, so that the hyperspectral video imaging system 2 has high flexibility and diversity, can meet the excitation requirements of different fluorescent materials at different wavelengths, and provide stable and reliable light source support for hyperspectral imaging; at the same time, the independent control of each laser light source 202 can more finely control the output of the light source and reduce the damage of the light source to the sample, thereby improving the quality and reliability of imaging.

[0035] The forward beam passes through the filter module 219 and the support cage rod 220 and enters the scanning galvanometer cage cube 221. After being reflected by the scanning galvanometer mirror 213 installed in the scanning galvanometer cage cube 221, it enters the slit sleeve 212 and becomes an elongated imaging ray after passing through the slit reflector 211. The imaging ray enters the sleeve grating 210 for diffraction to obtain the spectrum of the imaging ray. The diffracted spectrum changes the direction of the optical path through the imaging reflector 209 and propagates downward. It is focused by the first zoom lens 208 onto the two-dimensional plane of the high-speed CMOS 207.

[0036] Specifically, a visible light grating is installed in the sleeve grating 210 , which contains hundreds of scale lines per nanometer. The direction of the grating is perpendicular to the direction of the slit and parallel to the rotation direction of the scanning galvanometer 213 .

[0037] The galvanometer controller 206 controls the scanning galvanometer 213 by inputting different voltage signals to different rotation angles. When the galvanometer controller 206 continuously inputs a sinusoidal signal, the scanning galvanometer 213 scans the target along a specific direction. The scanned imaging rays pass through the slit sleeve 212 and slit mirror 211, producing a continuous spectrum. These spectra are continuously diffracted into spectral images and displayed on the high-speed CMOS 207.

[0038] The high-speed CMOS 207 stores each spectral image on the data acquisition card 204. Each image has M×N pixels and is continuously transmitted to the host computer 5. Within the host computer 5, the image is integrated along the width direction to obtain a column vector [N1], which can be used to obtain the pixels of the imaging ray. Each imaging ray is spliced ​​in a time series to form a target real image [N×T]. At the same time, the image is integrated along the height direction to obtain a one-dimensional horizontal vector [M1], which represents the complete spectrum of the target imaging ray. To obtain the spectrum of a specific point, simply process the xth row in the corresponding [M×N] to obtain the spectrum of the xth point on the imaging ray.

[0039] Figure 4The structure of the light source array system provided by the embodiment of the present invention is shown as follows: Figure 4 As shown, the light source array system 3 includes a light source bracket 301, a light source platform 302, a sapphire glass window 303, a light source array 304, and a laser 305. The light source bracket 301 is used to support and fix the optical platform 205 and the light source platform 302 in the hyperspectral video imaging system 2. The light source array 304 is fixedly mounted on the lower part of the optical platform 205. The laser 305 is arranged on the light source array 304. The sapphire glass window 303 is arranged on the light source platform 302. The arrangement position of the light source array 304 corresponds to the sapphire glass window.

[0040] In a specific embodiment, laser 305 receives laser light from laser source 202, which emits laser light at five wavelengths: 385 nm, 485 nm, 850 nm, 905 nm, and 1550 nm. When the laser light passes through sapphire glass window 303 and strikes imaging target 401, it excites imaging target 401 to emit fluorescence. Light reflected from imaging target 401 then passes through sapphire glass window 303 and returns to hyperspectral video imaging system 2.

[0041] Figure 5 The structure of the three-dimensional displacement system provided by the embodiment of the present invention is shown as follows: Figure 5 As shown, the three-dimensional displacement system 4 includes an imaging target 401, a stage 402, a stage 403, an X-axis scanning guide rail 404, an X-axis scanning motor, a Y-axis scanning motor 405, a Y-axis scanning guide rail 406, a Z-axis scanning platform 407, a Z-axis scanning platform leveling knob 408, a Z-axis scanning motor 409, a Z-axis scanning screw 410 and a Z-axis scanning guide rail 411.

[0042] The imaging target 401 is fixedly placed on the stage 402, and the stage is fixedly placed on the stage plate 403. The stage plate 403 is mounted on the X-axis scanning guide rail 404. The stage plate 403 can be moved on the X-axis scanning guide rail 404 by the X-axis scanning motor, so that the stage plate 403 can be moved relative to the X-axis direction; the X-axis scanning guide rail 404 is mounted on the Y-axis scanning guide rail 406, and the stage plate 403 can be moved relative to the Y-axis direction by the Y-axis scanning motor 405; the Y-axis scanning guide rail 406 is mounted on the Z-axis scanning platform 407, and the Z-axis scanning platform 407 can be balanced and calibrated by thread adjustment through the Z-axis scanning platform leveling knob 408. The Z-axis scanning platform 407 is mounted on the Z-axis scanning guide rail 411, and the Z-axis scanning platform 407 drives the Z-axis scanning screw 410 through the Z-axis scanning motor 409 to achieve up and down movement in the Z-axis direction. The three-dimensional displacement system 4 can realize precise movement and adjustment of the three-dimensional direction of the imaging target 401, thereby obtaining a clearer and more accurate imaging effect.

[0043] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0044] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

[0045] The above specific embodiments of the present invention do not constitute a limitation on the scope of protection of the present invention. Any other corresponding changes and modifications made based on the technical concept of the present invention should be included in the scope of protection of the claims of the present invention.

Claims

1. An animal fluorescence hyperspectral imager, characterized in that: The system comprises a hyperspectral video imaging system, a light source array system and a three-dimensional displacement system. The hyperspectral video imaging system comprises a light source intensity controller, a laser light source, an optical fiber, a data acquisition card, a galvanometer controller, a high-speed CMOS, a first imaging zoom lens, an imaging reflector, a sleeve grating, a slit reflector, a slit sleeve, a scanning galvanometer, a spectrometer cage cube, a sleeve, a turning lens barrel, a filter module, a support cage rod and a scanning galvanometer cage cube. The light source lamp array system includes a light source bracket, a light source platform, a sapphire glass window, a light source lamp array, and a laser. The light source bracket is used to support and fix the light source platform and the optical platform. The light source platform is fixed above the optical platform. The sapphire glass window is set on the light source platform. The light source lamp array is fixed on the lower surface of the optical platform, and the setting position of the light source lamp array corresponds to the setting position of the sapphire glass window. The laser is set on the light source lamp array and is used to receive the laser light emitted by the laser light source and irradiate the laser light onto the surface of the target sample after passing through the sapphire glass window. The laser light emitted by the laser light source is adjusted by the light source intensity controller and then transmitted to the laser through the optical fiber. The laser passes through the sapphire glass window and irradiates the surface of the target sample. The reflected light of the target sample passes through the sapphire glass window again and enters the turning lens barrel. The reflected light changes direction after passing through the turning lens barrel and enters the spectrometer cube through the sleeve. The scanning galvanometer is arranged in the scanning galvanometer cage cube, the galvanometer controller is used to control the rotation of the scanning galvanometer, and a visible light grating is arranged in the sleeve grating, and the direction of the visible light grating is perpendicular to the direction of the slit; the reflected light emitted from the spectroscopic cage cube passes through the filter module and the support cage rod in sequence and enters the scanning galvanometer cage cube, and the reflected light is then reflected by the scanning galvanometer and then passes through the slit sleeve and the slit reflector in sequence before being incident on the sleeve grating to form a diffraction spectrum. The diffraction spectrum passes through the imaging reflector and is focused by the first imaging zoom lens onto the two-dimensional plane of the high-speed CMOS sensor, and the high-speed CMOS stores the collected spectral image in the data acquisition card; The three-dimensional displacement system includes a stage, a loading plate, an X-axis scanning guide, an X-axis scanning motor, a Y-axis scanning guide, a Y-axis scanning motor, a Z-axis scanning platform, a Z-axis scanning motor, a Z-axis scanning guide and a Z-axis scanning lead screw. The stage is used to place an imaging target. The stage is fixedly arranged on the loading plate, the loading plate is mounted on the X-axis scanning guide rail, the X-axis scanning motor is used to drive the loading plate to move along the X-axis direction on the X-axis scanning guide rail; the X-axis scanning guide rail is mounted on the Y-axis scanning guide rail, the Y-axis scanning motor is used to drive the X-axis scanning guide rail and the loading plate to move along the Y-axis direction on the Y-axis scanning guide rail; the Y-axis scanning guide rail is fixedly mounted on the Z-axis scanning platform, the Z-axis scanning platform is mounted on the Z-axis scanning guide rail, the Z-axis scanning platform is connected to the Z-axis scanning lead screw, and the Z-axis scanning motor is used to drive the Z-axis scanning lead screw to drive the Z-axis scanning platform to move along the Z-axis direction on the Z-axis scanning guide rail.

2. The animal fluorescence hyperspectral imager according to claim 1, characterized in that: The system further includes an imaging CMOS and a second imaging zoom lens. A spectroscope is also provided in the spectroscopic cage cube for dividing the reflected light into two optical paths, a forward path and an upward path. The forward reflected light passes through the filter module and the support cage rods and then enters the scanning galvanometer cage cube. The upward reflected light passes through the second imaging zoom lens and then enters the imaging CMOS, and forms an image on the two-dimensional plane of the imaging CMOS sensor.

3. The animal fluorescence hyperspectral imager according to claim 1, characterized in that: The number of the laser light source is at least one, and the number of the light source intensity controllers is the same as the number of the laser light sources, which are used to individually control each laser light source to meet the excitation requirements of different fluorescent materials at different wavelengths.

4. The animal fluorescence hyperspectral imager according to claim 2, characterized in that: The focal length of the second imaging zoom lens is 18-35 nm.

5. The animal fluorescence hyperspectral imager according to claim 1, characterized in that: The three-dimensional displacement system further includes a Z-axis scanning platform leveling knob, and the Z-axis scanning platform leveling knob is used to adjust the Z-axis scanning platform through a thread to perform balance calibration.

Images