Portable single-photon calcium imaging microscopy device and method

Abstract

The present application relates to a portable single-photon calcium imaging camera device, comprising a dichroic lens, a shell, a beam splitter, an image receiver, an excitation light source and a self-focusing lens. The shell is provided with an inner cavity, a first opening, a second opening and a third opening. The first opening, the second opening and the third opening are all connected to the inner cavity, and the first opening and the second opening are opposite to each other. The image receiver, the excitation light source and the self-focusing lens are all arranged on the shell. The receiving surface of the image receiver is opposite to the first opening, the self-focusing lens is opposite to the second opening, and the excitation light source is opposite to the third opening. The beam splitter and the dichroic lens are both arranged in the inner cavity. The light input end of the dichroic lens is opposite to the light emitting end of the excitation light source, the beam splitter is opposite to the light output end of the dichroic lens, and the beam splitter is located between the image receiver and the self-focusing lens.

Description

Technical Field

[0001] The present invention relates to the field of imaging technology, and in particular to a portable single-photon calcium imaging microscopic device and method. Background Art

[0002] Brain science is an important research direction that is currently receiving international attention. In order to explain the neural basis of human cognition as the main body and core, and also to facilitate the study of the direct interaction between the brain and the outside world, neurons in the brain establish mutual connections through collective behavior, forming a variety of neural circuits to perform different functions. The research direction of neural circuits is to find the neuronal cells that constitute each neural circuit, study how these neurons produce communication interactions to form specific circuits and the life activities of the circuits themselves in the central nervous system. Many studies have confirmed that in the process of analyzing the regulatory mechanism of neural circuits under physiological or pathological conditions, it is necessary to accurately reflect the neuronal activity in brain tissue by real-time monitoring of changes in calcium ions, an important biosensor in neurons.

[0003] To solve the above problems, in the related art, the Chinese invention with patent number 201680077967.7 provides a system and method for simultaneous imaging and stimulation using a microscope system. Compared with an ordinary microscope system, the microscope system can have a relatively small size. The microscope can partially include an imaging light source and a stimulation light source. The light from the imaged light source and the stimulation light source can be spectrally separated to reduce the crosstalk between the stimulation light and the imaging light. In the above system and method, the stimulation light beam is calibrated by a condenser lens and a second condenser lens to focus the stimulation light into a light beam with little or no change in intensity within the beam area. The irradiated area of ​​the sample irradiated by the stimulation light will be reduced, resulting in a smaller beam of imaging light reflected by the irradiated area, and a smaller beam of imaging light guided by the second two-color element. At the same time, the barrel lens focuses the imaging light guided by the second two-color element onto the image sensor, resulting in a smaller image formed by the imaging sensor, which is inconvenient for staff to observe. Summary of the invention

[0004] Based on this, a portable single-photon calcium imaging microscope is provided to solve the problem in the related art that the irradiated area of ​​the sample by the stimulation light will be reduced, resulting in a smaller beam of imaging light reflected by the irradiated area, thereby resulting in a smaller beam of imaging light guided by the second two-color element, and further resulting in a smaller image formed by the imaging sensor.

[0005] A portable single-photon calcium imaging camera device comprises an astigmatism lens, a shell, a spectroscope, an image receiver, an excitation light source and a self-focusing lens. The shell is provided with an inner cavity, a first opening, a second opening and a third opening. The first opening, the second opening and the third opening are all connected to the inner cavity. The first opening and the second opening are opposite to each other. The image receiver, the excitation light source and the self-focusing lens are all arranged on the shell. The receiving surface of the image receiver is opposite to the first opening, the self-focusing lens is opposite to the second opening, the excitation light source is opposite to the third opening, the spectroscope and the lens are both arranged in the inner cavity, the light inlet end is opposite to the light emitting end of the excitation light source, the spectroscope is opposite to the light outlet end, and the spectroscope is located between the image receiver and the self-focusing lens.

[0006] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, a blue light filter layer is attached to the light-incoming surface of the light-incoming end.

[0007] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, the portable single-photon calcium imaging camera device also includes a fluorescent filter, and the fluorescent filter is arranged in the inner cavity, and the fluorescent filter is located between the spectroscope and the image receiver.

[0008] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, the portable single-photon calcium imaging camera device also includes an achromatic lens, which is arranged in the inner cavity and located between the beam splitter and the self-focusing lens.

[0009] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, the portable single-photon calcium imaging camera device also includes a focusing lens, and the focusing lens is arranged in the inner cavity, and the focusing lens is located between the beam splitter and the self-focusing lens.

[0010] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, the optical axis of the beam splitter, the optical axis of the lens and the optical axis of the self-focusing lens intersect with each other.

[0011] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, the portable single-photon calcium imaging camera device also includes a liquid lens, and the liquid lens is arranged in the inner cavity, and the liquid lens is located between the beam splitter and the self-focusing lens.

[0012] Preferably, in the above-mentioned portable single-photon calcium imaging camera device, the portable single-photon calcium imaging camera device also includes a battery, the battery is arranged on the housing, and the image receiver and the liquid lens are both electrically connected to the battery.

[0013] Based on the above-mentioned portable single-photon calcium imaging camera device, the present application also discloses a portable single-photon calcium imaging camera method, which is applicable to the above-mentioned portable single-photon calcium imaging camera device, and is characterized in that the portable single-photon calcium imaging camera method comprises:

[0014] The light emitted by the excitation light source is expanded by the astigmatism lens to obtain a first light beam;

[0015] Reflecting the first light beam through the self-focusing lens to the irradiation area of ​​the living body by the beam splitter;

[0016] The irradiated area reflects a second light beam which sequentially passes through the self-focusing lens and the beam splitter to the image receiver, and a brain fluorescent image is obtained through the image receiver.

[0017] Preferably, in the above-mentioned portable single-photon calcium imaging method, the step of expanding the light emitted by the excitation light source into the first light beam by the astigmatism lens comprises:

[0018] The blue light filter layer filters the light emitted by the excitation light source to obtain a third light beam;

[0019] The first light beam is obtained by expanding the third light beam through the divergent lens.

[0020] Preferably, in the above-mentioned portable single-photon calcium imaging method, the second light beam reflected by the irradiated area sequentially passes through the self-focusing lens and the beam splitter to the image receiver, and the brain fluorescence image is obtained through the image receiver, including:

[0021] Filtering the second light beam through the fluorescent filter to obtain a fourth light beam;

[0022] The brain fluorescence image is obtained by receiving the fourth light beam through the image receiver.

[0023] The technical solution adopted in this application can achieve the following beneficial effects:

[0024] In the portable single-photon calcium imaging camera device disclosed in the present application, it includes a diffuse lens, a shell, a spectroscope, an image receiver, an excitation light source and a self-focusing lens. The shell can be provided with an inner cavity, a first opening, a second opening and a third opening. The first opening, the second opening and the third opening can all be connected to the inner cavity. The first opening can be opposite to the second opening. The image receiver, the excitation light source and the self-focusing lens can all be arranged on the shell. The receiving surface of the image receiver can be opposite to the first opening, the self-focusing lens can be opposite to the second opening, the excitation light source can be opposite to the third opening, the spectroscope and the diffuse lens can all be arranged in the inner cavity, the light input end can be opposite to the light emitting end of the excitation light source, the spectroscope can be opposite to the light output end, and the spectroscope can be located between the image receiver and the self-focusing lens. In the above structure, since the diffuse lens expands the stimulation light, the irradiation area of ​​the stimulation light in the living brain is enlarged, thereby enlarging the beam of green fluorescence reflected by the living brain, thereby enlarging the size of the brain fluorescence image generated on the image receiver. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 An exploded diagram of a portable single-photon calcium imaging camera device disclosed in an embodiment of the present application;

[0026] Figure 2 It is a front view of the portable single-photon calcium imaging camera device disclosed in the embodiment of the present application;

[0027] Figure 3 for Figure 1 A cross-sectional view of

[0028] Figure 4 An exploded cross-sectional view of a housing disclosed in an embodiment of the present application;

[0029] Figure 5 A partial structural diagram of a portable single-photon calcium imaging camera device disclosed in an embodiment of the present application;

[0030] Figure 6 is the energy distribution diagram of the light source;

[0031] Figure 7 is the expected energy distribution diagram of the light emitted by the light source;

[0032] Figure 8 An image taken by irradiating the brain using the astigmatism lens disclosed in the embodiment of the present application;

[0033] Fig. 9 This is an image taken without using the astigmatism lens disclosed in the embodiment of the present application to illuminate the brain.

[0034] Among them: a diffuser lens 100, a light input end 110, a light output end 120, a blue light filter layer 130, a shell 200, an inner cavity 210, a first opening 220, a second opening 230, a third opening 240, a spectroscope 300, an image receiver 400, an excitation light source 500, a self-focusing lens 600, an achromatic lens 700, a focusing lens 800, a liquid lens 900, and a fluorescent filter 1000. DETAILED DESCRIPTION

[0035] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thoroughly understood.

[0036] It should be noted that when an element is referred to as being "disposed on" another element, it may be directly on the other element or there may be a central element. When an element is considered to be "connected to" another element, it may be directly connected to the other element or there may be a central element at the same time. The terms "vertical", "horizontal", "left", "right", "top", "bottom", "bottom end", "top end" and similar expressions used herein are for illustrative purposes only and do not represent the only implementation method.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0038] Please refer to Figures 1 to 5 The present application discloses a portable single-photon calcium imaging camera device, which includes a dichroic lens 100, a housing 200, a beam splitter 300, an image receiver 400, an excitation light source 500 and a self-focusing lens 600.

[0039] The shell 200 can provide an installation base for the astigmatism lens 100, the spectroscope 300, the image receiver 400, the excitation light source 500 and the self-focusing lens 600. The excitation light source 500 can emit stimulation light, which can stimulate the GCaMP virus in the living brain to emit green fluorescence. The self-focusing lens 600 has focusing and imaging functions.

[0040] Specifically, the shell 200 may be provided with an inner cavity 210, a first opening 220, a second opening 230 and a third opening 240, the first opening 220, the second opening 230 and the third opening 240 may all be connected to the inner cavity 210, the first opening 220 may be opposite to the second opening 230, the image receiver 400, the excitation light source 500 and the self-focusing lens 600 may all be arranged on the shell 200, the receiving surface of the image receiver 400 may be opposite to the first opening 220, the self-focusing lens 600 may be opposite to the second opening 230, the excitation light source 500 may be opposite to the third opening 240, the beam splitter 300 and the astigmatism lens 100 may all be arranged in the inner cavity 210, the light input end 110 may be opposite to the light emitting end of the excitation light source 500, the beam splitter 300 may be opposite to the light output end 120, and the beam splitter 300 may be located between the image receiver 400 and the self-focusing lens 600.

[0041] During the use of the portable single-photon calcium imaging camera device, the excitation light source 500 emits stimulation light to the light input end 110 of the astigmatism lens 100, and the stimulation light is emitted from the light output surface of the light output end 120. As can be seen from the above, the light output surface of the light output end 120 is a convex surface formed by the rotation of multiple light output points on a Cartesian coordinate system, so that multiple light rays emitted from multiple light output points can be evenly and diffusely emitted to the spectroscope 300, and the spectroscope 300 reflects the stimulation light to the self-focusing lens 600. The stimulation light passes through the self-focusing lens 600 to the living brain. The GCaMP virus in the living brain is stimulated by the stimulation light and emits green fluorescence, which passes through the self-focusing lens 600 and the spectroscope 300 to the image receiver 400 in sequence, thereby generating a brain fluorescence image.

[0042] The portable single-photon calcium imaging camera device disclosed in the present application includes a diffraction lens 100, a housing 200, a beam splitter 300, an image receiver 400, an excitation light source 500 and a self-focusing lens 600. The housing 200 may be provided with an inner cavity 210, a first opening 220, a second opening 230 and a third opening 240. The first opening 220, the second opening 230 and the third opening 240 may all be connected to the inner cavity 210. The first opening 220 may be opposite to the second opening 230. The image receiver 400, the excitation light source 500 and the self-focusing lens 600 may be connected to the inner cavity 210. The focusing lens 600 can be arranged on the housing 200, the receiving surface of the image receiver 400 can be opposite to the first opening 220, the self-focusing lens 600 can be opposite to the second opening 230, the excitation light source 500 can be opposite to the third opening 240, the beam splitter 300 and the diffusing lens 100 can be arranged in the inner cavity 210, the light input end 110 can be opposite to the light emitting end of the excitation light source 500, the beam splitter 300 can be opposite to the light output end 120, and the beam splitter 300 can be located between the image receiver 400 and the self-focusing lens 600. In the above structure, since the diffusing lens 100 expands the stimulation light, the irradiation area of ​​the stimulation light in the living brain is expanded, thereby expanding the green fluorescence beam reflected by the living brain, thereby expanding the size of the brain fluorescence image generated on the image receiver 400.

[0043] At the same time, the stimulation light beam is irradiated to the living brain uniformly and expanded, that is, the GCaMP virus in the living brain is irradiated by the stimulation light beam with uniform light intensity, and the green fluorescence generated by the GCaMP virus in the living brain is also uniform, thereby improving the clarity of the brain fluorescence image.

[0044] In an embodiment of the present application, the portable single-photon calcium imaging camera device may further include a fluorescent filter 1000. Specifically, the fluorescent filter 1000 may be disposed in the inner cavity 210, and the fluorescent filter 1000 may be located between the spectroscope 300 and the image receiver 400, wherein the fluorescent filter 1000 is used to allow only green fluorescence to pass, thereby preventing light of other colors from being transmitted to the image receiver 400 and interfering with the generation of brain fluorescent images.

[0045] In the embodiment of the present application, the portable single-photon calcium imaging camera device may further include an achromatic lens 700. Specifically, the achromatic lens 700 may be disposed in the inner cavity 210. The achromatic lens 700 may be located between the beam splitter 300 and the self-focusing lens 600. The achromatic lens 700 is used to eliminate the chromatic aberration and spherical aberration of the green fluorescence or the stimulation light. Chromatic aberration and spherical aberration will occur during the transmission of the stimulation light and the green fluorescence. The chromatic aberration is caused by the difference in dispersion and refractive index of light of different wavelengths in the glass, which results in different focal points for light of different wavelengths. The spherical aberration is that the light beam emitted by the object point on the axis, after passing through the optical system, intersects the optical axis at different positions with the light beam at different angles with the optical axis, thus forming a circular diffuse spot on the image plane. It can be seen that the above structure can improve the resolution of the image receiver 400 through the achromatic lens 700.

[0046] In an embodiment of the present application, the portable single-photon calcium imaging camera device may further include a focusing lens 800. Specifically, the focusing lens 800 may be disposed in the inner cavity 210. The focusing lens 800 may be located between the beam splitter 300 and the self-focusing lens 600. The focusing lens 800 may adjust small deviations such as eccentricity of the green fluorescence or stimulation light, so that most of the green fluorescence or stimulation light may pass through the focusing lens 800 and reach a designated position.

[0047] In an embodiment of the present application, the portable single-photon calcium imaging camera device may further include a liquid lens 900 and a controller. Specifically, the liquid lens 900 may be disposed in the inner cavity 210, the controller may be disposed on the housing 200, the liquid lens 900 may be located between the beam splitter 300 and the self-focusing lens 600, the controller may be electrically connected to the liquid lens 900, and when the liquid lens 900 is in a powered-on state, the controller may control the deformation of the liquid lens 900, wherein the liquid lens 900 uses the liquid as an astigmatism lens 100 to change the focal length by changing the curvature of the liquid, thereby realizing automatic focusing and zooming of the portable single-photon calcium imaging camera device, thereby improving the image effect captured by the portable single-photon calcium imaging camera device.

[0048] In an embodiment of the present application, the portable single-photon calcium imaging camera device may further include a battery. Specifically, the battery may be disposed on the housing 200, and the image receiver 400 and the liquid lens 900 may be electrically connected to the battery, thereby enabling the portable single-photon calcium imaging camera device to be disconnected from the power supply connected via the electrical connector, thereby enabling the portable single-photon calcium imaging camera device to facilitate free movement of the living body.

[0049] In the embodiment of the present application, a blue light filter layer 130 may be attached to the light input surface of the light input end 110. Since the blue light band in the stimulation light can excite the GCaMP virus to produce green fluorescence, the blue light filter layer 130 can only allow the blue light in the stimulation light to pass through, thereby preventing light of other color bands from irradiating the living brain and causing adverse effects.

[0050] The disclosed astigmatism lens 100 establishes a Cartesian coordinate system with the position of the light source as the origin, and a fitting curve fitted by a plurality of lens profile surface points is rotated to form a closed three-dimensional model, and the closed three-dimensional model is the astigmatism lens 100. The coordinate positions of the plurality of lens profile surface points in the Cartesian coordinate system can be obtained by the following formula:

[0051]

[0052] Wherein, x is the X-axis coordinate value of any one of the plurality of lens surface points in the Cartesian coordinate system, y is the Y-axis coordinate value of any one of the plurality of lens surface points in the Cartesian coordinate system, N ix is the normal component of any one of the multiple lens surface points in the X-axis direction on the Cartesian coordinate system, N iy is the normal component of any one of the multiple lens surface points in the Y-axis direction on the Cartesian coordinate system, I (i+1)x is the unit vector of the incident light in the Cartesian coordinate system that matches any one of the multiple lens surface points, in the X-axis direction, (i+1)y is the unit vector of the Y-axis direction of the incident light in the Cartesian coordinate system that matches any one of the multiple lens surface points, θ j is the position of any one of the multiple lens surface points, P ix is the X-axis coordinate value of the incident point of the incident light matched with any one of the multiple lens surface points on the Cartesian coordinate system, P iy It is the Y-axis coordinate value of the incident point of the incident light matched with any one of the multiple lens surface points on the Cartesian coordinate system.

[0053] The radiance distribution of the light source is known, that is, the energy distribution of the light emitted by the light source is known. If the light source is regarded as a point, its energy distribution can be roughly approximated as an axisymmetric distribution, such as Figure 6 and Figure 7 As shown, Ω is the energy distribution of the light source, and S is regarded as the expected energy distribution of the light emitted by the light source. Ω and S are divided into energy grids respectively. For the division of Ω, since the energy distribution is known, the energy in each grid can be guaranteed to be equal during the division; for the division of S, the area in each grid must also be guaranteed to be equal during the division.

[0054] The above grid division is used to calculate the profile of the astigmatism lens 100. The light energy distribution effect that the astigmatism lens 100 can achieve is the energy distribution on S. Through the above energy division, the boundaries of each grid of Ω and S are determined. Therefore, the incident light vector I i We can determine them one by one, and use Snell's law to calculate the normal vector N corresponding to the point i , normal vector N i It is the surface normal vector of the point that can refract the incident light to the specified position. Multiple lens surface points are calculated one by one through the above formula by iteratively using the normal vector.

[0055] During the use of the astigmatism lens 100, the stimulation light source needs to illuminate the illumination area of ​​the living brain, and the multiple light rays emitted by the stimulation light source are refracted by the astigmatism lens 100. Since the astigmatism lens 100 is a closed three-dimensional model, the closed three-dimensional model is formed by rotating the fitting curve of the surface points of the lens profile, so that the multiple light rays emitted by the stimulation light source can be refracted by the astigmatism lens 100 and emitted uniformly and diffusely, so that the illumination area of ​​the multiple light rays on the living brain becomes larger, and by Figure 8 and Fig. 9 As shown, Figure 8 is an image of brain neurons captured through the astigmatism lens 100. Fig. 9 is a brain neuron image taken without being illuminated by the astigmatism lens 100, wherein: Figure 8 and Fig. 9 Images at the same magnification.

[0056] In the embodiment of the present application, the astigmatism lens 100 may be an acrylic lens. Specifically, the acrylic lens is made of acrylic material. The acrylic material has the advantages of light weight and high light transmittance. Meanwhile, the acrylic material has good processing performance.

[0057] The present application also discloses a portable single-photon calcium imaging camera method, which is applicable to the above-mentioned portable single-photon calcium imaging camera device. The disclosed portable single-photon calcium imaging camera method includes:

[0058] Step 101, expanding the light emitted by the excitation light source 500 through the astigmatism lens 100 to obtain a first light beam;

[0059] In this step, the excitation light source 500 emits stimulation light to the light input end 110 of the astigmatism lens 100, and the stimulation light is emitted from the light output surface of the light output end 120. The light output surface of the light output end 120 is a convex surface formed by rotating a curve formed by multiple light output points on a Cartesian coordinate system, so that the multiple light rays emitted by the multiple light output points can form a diffuse and uniform first light beam.

[0060] Step 102, the first light beam is reflected by the beam splitter 300 and passes through the self-focusing lens 600 to the irradiation area of ​​the living body;

[0061] Step 103 , the second light beam reflected from the irradiated area passes through the self-focusing lens 600 and the beam splitter 300 in sequence to the image receiver 400 , and a brain fluorescent image is obtained through the image receiver 400 .

[0062] In this step, the GCaMP virus in the living brain is stimulated by irradiation to emit a second light beam which passes through the self-focusing lens 600 and the beam splitter 300 in sequence to the image receiver 400, thereby generating a brain fluorescent image.

[0063] In the embodiment of the present application, expanding the light emitted by the excitation light source 500 into a first light beam by the diffuser lens 100 may include:

[0064] Step 201, filtering the light emitted by the excitation light source 500 through the blue light filter layer 130 to obtain a third light beam;

[0065] In this step, since the blue light band in the stimulation light can excite the GCaMP virus to produce green fluorescence, the blue light filter layer 130 can only allow the blue light in the stimulation light to pass through to form a third light beam, thereby preventing light of other color bands from irradiating the living brain and causing adverse effects.

[0066] Step 202 : Expand the third light beam through the astigmatism lens 100 to obtain the first light beam.

[0067] In the embodiment of the present application, the second light beam reflected by the irradiation area sequentially passes through the self-focusing lens 600 and the beam splitter 300 to the image receiver 400, and the brain fluorescence image is obtained through the image receiver 400, which may include:

[0068] Step 301, filtering the second light beam through the fluorescent filter 1000 to obtain a fourth light beam;

[0069] In this step, the fluorescent filter 1000 can only allow green fluorescence to pass through to form the fourth light beam, thereby preventing light of other colors from being transmitted to the image receiver 400 and causing interference with the generation of brain fluorescent images.

[0070] Step 302 : Receive a fourth light beam through the image receiver 400 to obtain a brain fluorescent image.

[0071] The technical features of the above embodiments may be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0072] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A portable single-photon calcium imaging camera device, characterized in that: The invention comprises an astigmatism lens, a shell, a beam splitter, an image receiver, an excitation light source and a self-focusing lens. The shell is provided with an inner cavity, a first opening, a second opening and a third opening. The first opening, the second opening and the third opening are all connected to the inner cavity. The first opening and the second opening are opposite to each other. The image receiver, the excitation light source and the self-focusing lens are all arranged on the shell. The receiving surface of the image receiver is opposite to the first opening. The self-focusing lens is opposite to the second opening. The excitation light source is opposite to the third opening. The beam splitter and the astigmatism lens are all arranged in the inner cavity. The light input end of the astigmatism lens is opposite to the light emitting end of the excitation light source. The beam splitter is opposite to the light output end of the astigmatism lens. The beam splitter is located between the image receiver and the self-focusing lens. The astigmatism lens establishes a Cartesian coordinate system with the position of the light source as the origin. A fitting curve fitted by a plurality of lens profile surface points is formed by rotation to form a closed three-dimensional model. The closed three-dimensional model is an astigmatism lens. The coordinate positions of the plurality of lens profile surface points in the Cartesian coordinate system can be obtained by the following formula: Wherein, x is the X-axis coordinate value of any one of the plurality of lens surface points in the Cartesian coordinate system, y is the Y-axis coordinate value of any one of the plurality of lens surface points in the Cartesian coordinate system, N ix is the normal component of any one of the multiple lens surface points in the X-axis direction on the Cartesian coordinate system, N iy is the normal component of any one of the multiple lens surface points in the Y-axis direction on the Cartesian coordinate system, I (i+1)x is the unit vector of the incident light in the Cartesian coordinate system that matches any one of the multiple lens surface points, in the X-axis direction, (i+1)y is the unit vector of the Y-axis direction of the incident light in the Cartesian coordinate system that matches any one of the multiple lens surface points, θ j is the position of any one of the multiple lens surface points, P ix is the X-axis coordinate value of the incident point of the incident light matched with any one of the multiple lens surface points on the Cartesian coordinate system, P iy It is the Y-axis coordinate value of the incident point of the incident light matched with any one of the multiple lens surface points on the Cartesian coordinate system.

2. The portable single-photon calcium imaging camera device according to claim 1, characterized in that: A blue light filter layer is attached to the light input surface of the light input end.

3. The portable single-photon calcium imaging camera device according to claim 1, characterized in that: The portable single-photon calcium imaging camera device further comprises a fluorescent filter, which is arranged in the inner cavity and located between the beam splitter and the image receiver.

4. The portable single-photon calcium imaging camera device according to claim 1, characterized in that: The portable single-photon calcium imaging camera device further comprises an achromatic lens, which is arranged in the inner cavity and located between the beam splitter and the self-focusing lens.

5. The portable single-photon calcium imaging camera device according to claim 1, characterized in that: The portable single-photon calcium imaging camera device further comprises a focusing lens, which is arranged in the inner cavity and located between the beam splitter and the self-focusing lens.

6. The portable single-photon calcium imaging camera device according to claim 1, characterized in that: The optical axis of the beam splitter, the optical axis of the lens and the optical axis of the self-focusing lens intersect with each other.

7. The portable single-photon calcium imaging camera device according to claim 1, characterized in that: The portable single-photon calcium imaging camera device further comprises a liquid lens, which is arranged in the inner cavity and located between the beam splitter and the self-focusing lens.

8. The portable single-photon calcium imaging camera device according to claim 7, characterized in that: The portable single-photon calcium imaging camera device further comprises a battery, which is arranged on the housing, and the image receiver and the liquid lens are both electrically connected to the battery.

9. A portable single-photon calcium imaging camera method, applicable to the portable single-photon calcium imaging camera device according to any one of claims 1 to 8, characterized in that: The portable single-photon calcium imaging method comprises: The first light beam is obtained by expanding the light emitted by the excitation light source component through the astigmatism lens: the excitation light source component emits stimulation light to the light inlet end of the astigmatism lens, and the stimulation light is emitted from the light outlet surface of the light outlet end, and the light outlet surface of the light outlet end is a convex surface formed by rotating a curve formed by multiple light outlet points on a Cartesian coordinate system, so that multiple light rays emitted by multiple light outlet points can form a diffuse and uniform first light beam; Reflecting the first light beam through the self-focusing lens to the irradiation area of ​​the living body by the beam splitter; The irradiated area reflects a second light beam which sequentially passes through the self-focusing lens and the beam splitter to the image receiver, and a brain fluorescent image is obtained through the image receiver.

10. The portable single-photon calcium imaging method according to claim 9, characterized in that: The light-incoming surface of the light-incoming end is provided with a blue light filter layer, and the light emitted by the excitation light source is expanded into the first light beam through the diffuser lens, including: The blue light filter layer filters the light emitted by the excitation light source to obtain a third light beam; The first light beam is obtained by expanding the third light beam through the divergent lens.

11. The portable single-photon calcium imaging method according to claim 9, characterized in that: The portable single-photon calcium imaging camera device further includes a fluorescent filter, which is disposed in the inner cavity and located between the beam splitter and the image receiver. The second light beam reflected by the irradiated area sequentially passes through the self-focusing lens and the beam splitter to the image receiver, and the brain fluorescent image is obtained through the image receiver, including: Filtering the second light beam through the fluorescent filter to obtain a fourth light beam; The brain fluorescence image is obtained by receiving the fourth light beam through the image receiver.

Images