A dual-channel visible light-near-infrared second-window light confocal microscope

Abstract

The present invention relates to a dual-channel visible light-near-infrared second-region light confocal microscope, comprising: a visible light source for emitting a first visible light; a visible light scanning module for transmitting the first visible light and scanning a sample at a first speed or a second speed; a near-infrared second-region light source for emitting a first near-infrared second-region light; a near-infrared second-region light scanning module for transmitting the first near-infrared second-region light and scanning the sample; a focusing module for focusing the first visible light or the first near-infrared second-region light on a point on the sample to excite a fluorescent substance on the sample to obtain a first fluorescence or a second fluorescence; a first acquisition module for acquiring the first fluorescence; and a second acquisition module for acquiring the second fluorescence. The excitation light in the present invention can be visible lights of multiple different wavelength bands and near-infrared second-region lights of multiple different wavelength bands, and visible light scanning of multiple different wavelength bands and near-infrared second-region light scanning of multiple different wavelength bands can be realized.

Description

Technical Field

[0001] The present invention relates to the technical field of microscopes, and particularly to a dual-channel visible light-near-infrared II light confocal microscope. Background Art

[0002] A confocal laser scanning microscope (CLSM) is one of the most advanced cell biomedical analysis instruments in modern times. It is based on fluorescence microscopy imaging and is equipped with a laser scanning device. It uses laser fluorescence probes in bands such as ultraviolet light or visible light lasers, and uses a computer for image processing. It can not only observe fixed cells and tissue sections, but also observe and detect the structure, molecules, and ions of living cells in real time and dynamically. Confocal laser scanning microscopy technology has been used in research such as cell morphology localization, three-dimensional structure reconstruction, and dynamic change processes, and provides practical research means such as quantitative fluorescence measurement and quantitative image analysis. Combined with other related biotechnology, it has been widely used in molecular cell biology fields such as morphology, physiology, immunology, and genetics.

[0003] At present, the research on photoluminescence by confocal microscopes mainly focuses on the visible light band range and the near-infrared I band (NIR I: 400 nm - 900 nm). However, biological tissues have strong scattering and absorption of fluorescence in the visible light and near-infrared I bands, so that the imaging depth corresponding to the optical microscopy imaging technology based on the above bands is often in the micron order. Therefore, it is impossible to image biological tissues located deeper in the tissue, and it is difficult to meet the increasingly urgent deep imaging requirements. Summary of the Invention

[0004] The purpose of the present invention is to provide a dual-channel visible light-near-infrared II light confocal microscope to solve at least one of the problems existing in the prior art.

[0005] To achieve the above purpose, the present invention provides the following solutions:

[0006] A dual-channel visible light-near-infrared II light confocal microscope, the dual-channel visible light-near-infrared II light confocal microscope includes:

[0007] A visible light source for emitting first visible light; the first visible light is visible light of one band of visible light or visible light obtained by combining multiple different bands of visible light;

[0008] A visible light scanning module for transmitting the first visible light and scanning a sample at a first speed or a second speed; the first speed is higher than the second speed;

[0009] A near-infrared second-region light source for emitting first near-infrared second-region light; the first near-infrared second-region light is near-infrared second-region light of one band or near-infrared second-region light after combining near-infrared second-region lights of multiple different bands;

[0010] A near-infrared second-region light scanning module for transmitting the first near-infrared second-region light and scanning a sample;

[0011] A focusing module for focusing the first visible light on a point on the sample to excite a fluorescent substance on the sample to obtain first fluorescence; and also for focusing the first near-infrared second-region light on a point on the sample to excite a fluorescent substance on the sample to obtain second fluorescence;

[0012] A first acquisition module for acquiring the first fluorescence;

[0013] A second acquisition module for acquiring the second fluorescence.

[0014] Optionally, the visible light source includes a first visible light source, a second visible light source, a first window plate, a second window plate, a first reflector, a first dichroic mirror, a first plano-convex lens, and a first optical fiber;

[0015] The first visible light source is used for emitting second visible light; the second visible light source is used for emitting third visible light; the second visible light and the third visible light have different bands;

[0016] The first window plate and the first reflector are sequentially arranged on the propagation path of the second visible light; the first window plate is used for deflecting the second visible light; the first reflector is used for reflecting the deflected second visible light;

[0017] The second window plate and the first dichroic mirror are sequentially arranged on the propagation path of the third visible light; the first dichroic mirror is also arranged on the propagation path of the deflected second visible light; the second window plate is used for deflecting the third visible light; the first dichroic mirror is used for reflecting the deflected third visible light and transmitting the deflected second visible light, and also combining the deflected third visible light and the deflected second visible light into a beam of light, denoted as fourth visible light;

[0018] The first plano-convex lens is arranged on the propagation path of the fourth visible light; the first plano-convex lens is used for focusing and coupling the fourth visible light into the first optical fiber.

[0019] Optionally, the visible light scanning module includes a visible light source optical fiber, a first collimation module, a resonant galvanometer, a first y-axis galvanometer mirror, a first 4f system, a first x-axis galvanometer mirror, a visible light scanning lens, and a visible light band multi-bandpass dichroic mirror sequentially arranged on the propagation path of the first visible light;

[0020] The visible light source optical fiber is used to confine the first visible light to a point light source and transmit the first visible light;

[0021] The first collimation module is used to collimate the first visible light;

[0022] The resonant galvanometer and the first y-axis galvanometer mirror are used to scan the sample at a first speed;

[0023] The first y-axis galvanometer mirror and the first x-axis galvanometer mirror are used to scan the sample at a second speed, and the first speed is higher than the second speed;

[0024] The first 4f system is used to collimate and optimize the optical path;

[0025] The visible light scanning lens is used to adapt to the wavelength of the first visible light and reduce aberration;

[0026] The visible light band multi-bandpass dichroic mirror is used to reflect the first visible light.

[0027] Optionally, the second near-infrared light source includes a first second near-infrared light source, a second second near-infrared light source, a third window plate, a fourth window plate, a second reflector, a third dichroic mirror, a second plano-convex lens, and a second optical fiber;

[0028] The first second near-infrared light source is used to emit a second second near-infrared light; the second second near-infrared light source is used to emit a third second near-infrared light; the second second near-infrared light and the third second near-infrared light have different wavelength bands;

[0029] The third window plate and the second reflector are sequentially arranged on the propagation path of the second second near-infrared light; the third window plate is used to deflect the second second near-infrared light; the second reflector is used to reflect the deflected second second near-infrared light;

[0030] The fourth window plate and the third dichroic mirror are sequentially arranged on the propagation path of the third second near-infrared light; the third dichroic mirror is also arranged on the propagation path of the deflected second second near-infrared light; the fourth window plate is used to deflect the third second near-infrared light; the third dichroic mirror is used to reflect the deflected third second near-infrared light and transmit the deflected second second near-infrared light, and also combines the deflected third second near-infrared light and the deflected second second near-infrared light into a beam of light, denoted as the fourth second near-infrared light;

[0031] The second plano-convex lens is disposed on the propagation path of the fourth second near-infrared region light; the second plano-convex lens is configured to focus and couple the fourth second near-infrared region light into the second optical fiber.

[0032] Optionally, the second near-infrared region light scanning module includes a near-infrared light source optical fiber, a second collimation module, a near-infrared band multi-band dichroic mirror, a second x-axis galvanometric mirror, a second 4f system, a second y-axis galvanometric mirror, and an infrared light scanning lens, which are sequentially disposed on the propagation path of the first second near-infrared region light;

[0033] The near-infrared light source optical fiber is configured to confine the first second near-infrared region light to a point light source and transmit the first second near-infrared region light;

[0034] The second collimation module is configured to collimate the first second near-infrared region light;

[0035] The near-infrared band multi-band dichroic mirror is configured to reflect the first second near-infrared region light;

[0036] The second x-axis galvanometric mirror and the second y-axis galvanometric mirror are configured to scan the sample;

[0037] The second 4f system is configured to collimate and optimize the optical path;

[0038] The infrared light scanning lens is configured to adapt to the wavelength of the first second near-infrared region light and reduce aberration.

[0039] Optionally, the focusing module includes a long-pass dichroic mirror, a tube lens, a z-axis piezoelectric displacement stage, an objective lens, and a sample stage;

[0040] The long-pass dichroic mirror is disposed on the propagation path of the first visible light reflected by the visible light band multi-band dichroic mirror and the first second near-infrared region light transmitted by the infrared light scanning lens; the long-pass dichroic mirror is configured to reflect the first visible light and transmit the first second near-infrared region light;

[0041] The first visible light or the first second near-infrared region light sequentially passes through the long-pass dichroic mirror, the tube lens, the z-axis piezoelectric displacement stage, and the objective lens and then is focused on a point on the sample on the sample stage;

[0042] The sample stage is configured to carry the sample and adjust the scanning area of the x-y plane of the sample;

[0043] The z-axis piezoelectric displacement stage is configured to adjust the z-axis height of the objective lens;

[0044] The objective lens is used to focus the first visible light on a point on the sample, excite the fluorescent substance on the sample, and obtain the first fluorescence; the objective lens is also used to focus the first second near-infrared light on a point on the sample, excite the fluorescent substance on the sample, and obtain the second fluorescence;

[0045] The tube lens is used to correct the phase difference of the objective lens.

[0046] Optionally, the first acquisition module includes a first Fresnel lens, a first motorized pinhole wheel, a first multimode optical fiber, and a first acquisition unit; the objective lens, the z-axis piezoelectric displacement stage, the tube lens, the long-pass dichroic mirror, and the visible light band multi-band dichroic mirror are also sequentially arranged on the propagation path of the first fluorescence; the long-pass dichroic mirror is also used to reflect the first fluorescence; the visible light band band-pass dichroic mirror is also used to transmit the first fluorescence;

[0047] The first Fresnel lens is arranged on the propagation path of the first fluorescence transmitted by the visible light band band-pass dichroic mirror; the first Fresnel lens is used to focus the first fluorescence on the first motorized pinhole wheel;

[0048] The first motorized pinhole wheel is used to adjust the pinhole size and reduce external fluorescence noise;

[0049] One end of the first multimode optical fiber is connected to the first motorized pinhole wheel; the first multimode optical fiber is used to transmit the first fluorescence;

[0050] The first acquisition unit is connected to the other end of the first multimode optical fiber; the first acquisition unit is used to acquire the first fluorescence.

[0051] Optionally, the second acquisition module includes a fourth Fresnel lens, a second motorized pinhole wheel, a second multimode optical fiber, and a second acquisition unit; the objective lens, the z-axis piezoelectric displacement stage, the tube lens, the long-pass dichroic mirror, and the infrared light scanning lens are also sequentially arranged on the propagation path of the second fluorescence; the long-pass dichroic mirror and the near-infrared band multi-band dichroic mirror are also both used to transmit the second fluorescence;

[0052] The fourth Fresnel lens is arranged on the propagation path of the second fluorescence transmitted by the near-infrared band multi-band dichroic mirror; the fourth Fresnel lens is used to focus the second fluorescence on the second motorized pinhole wheel;

[0053] The second motorized pinhole wheel is used to adjust the pinhole size and reduce external fluorescence noise;

[0054] One end of the second multimode optical fiber is connected to the second motorized pinhole wheel; the second multimode optical fiber is used to transmit the second fluorescence;

[0055] The second acquisition unit is connected to the other end of the second multimode optical fiber; the second acquisition unit is configured to acquire the second fluorescence.

[0056] Optionally, the first acquisition unit includes a second dichroic mirror, a first filter, a second Fresnel lens, a first photomultiplier tube, a first signal amplifier, a second filter, a third Fresnel lens, a second photomultiplier tube, and a second signal amplifier;

[0057] The second dichroic mirror is disposed on the propagation path of the first fluorescence; the second dichroic mirror is configured to split the first fluorescence, transmit the first-band fluorescence in the first fluorescence and reflect the second-band fluorescence in the first fluorescence; the first-band fluorescence and the second-band fluorescence respectively correspond to the bands of the visible light emitted by the visible light source;

[0058] The first filter, the second Fresnel lens, and the first photomultiplier tube are sequentially disposed on the propagation path of the first-band fluorescence transmitted by the second dichroic mirror; the first filter is configured to filter the first-band fluorescence; the second Fresnel lens is configured to focus the first-band fluorescence into the first photomultiplier tube;

[0059] The first photomultiplier tube is configured to convert the first-band fluorescence focused by the second Fresnel lens into a first electrical signal;

[0060] One end of the first signal amplifier is connected to the first photomultiplier tube, and the first signal amplifier is configured to amplify the first electrical signal;

[0061] The second filter, the third Fresnel lens, and the second photomultiplier tube are sequentially disposed on the propagation path of the second-band fluorescence reflected by the second dichroic mirror; the second filter is configured to filter the second-band fluorescence; the third Fresnel lens is configured to focus the second-band fluorescence into the second photomultiplier tube;

[0062] The second photomultiplier tube is configured to convert the second-band fluorescence focused by the third Fresnel lens into a second electrical signal;

[0063] One end of the second signal amplifier is connected to the second photomultiplier tube, and the second signal amplifier is configured to amplify the second electrical signal;

[0064] The other end of the first signal amplifier is connected to the other end of the second signal amplifier; the other end of the first signal amplifier or the other end of the second signal amplifier is further connected to an external processor.

[0065] Optionally, the second acquisition unit includes a fourth dichroic mirror, a third filter, a fifth Fresnel lens, a third photomultiplier tube, a third signal amplifier, a fourth filter, a sixth Fresnel lens, a fourth photomultiplier tube, and a fourth signal amplifier;

[0066] The fourth dichroic mirror is disposed on the propagation path of the second fluorescence; the fourth dichroic mirror is configured to split the second fluorescence, transmit the third-band fluorescence in the second fluorescence, and reflect the fourth-band fluorescence in the second fluorescence; the third-band fluorescence and the fourth-band fluorescence respectively correspond to the bands of the second near-infrared light emitted by the second near-infrared light source;

[0067] The third filter, the fifth Fresnel lens, and the third photomultiplier tube are sequentially disposed on the propagation path of the third-band fluorescence transmitted by the fourth dichroic mirror; the third filter is configured to filter the third-band fluorescence; the fifth Fresnel lens is configured to focus the third-band fluorescence into the third photomultiplier tube;

[0068] The third photomultiplier tube is configured to convert the third-band fluorescence focused by the fifth Fresnel lens into a third electrical signal;

[0069] One end of the third signal amplifier is connected to the third photomultiplier tube, and the third signal amplifier is configured to amplify the third electrical signal;

[0070] The fourth filter, the sixth Fresnel lens, and the fourth photomultiplier tube are sequentially disposed on the propagation path of the fourth-band fluorescence reflected by the fourth dichroic mirror; the fourth filter is configured to filter the fourth-band fluorescence; the sixth Fresnel lens is configured to focus the fourth-band fluorescence into the fourth photomultiplier tube;

[0071] The fourth photomultiplier tube is configured to convert the fourth-band fluorescence focused by the sixth Fresnel lens into a fourth electrical signal;

[0072] One end of the fourth signal amplifier is connected to the fourth photomultiplier tube, and the fourth signal amplifier is configured to amplify the fourth electrical signal;

[0073] The other end of the third signal amplifier is connected to the other end of the fourth signal amplifier; the other end of the third signal amplifier or the other end of the fourth signal amplifier is further connected to an external processor

[0074] According to the specific embodiments provided by the present invention, the present invention discloses the following technical effects:

[0075] The present invention discloses a dual-channel visible light-near-infrared II light confocal microscope, which includes a visible light source for emitting first visible light; the first visible light is visible light of one wavelength band of visible light or visible light after combining multiple different wavelength bands of visible light; a visible light scanning module for transmitting the first visible light and scanning a sample at a first speed or a second speed; the first speed is higher than the second speed; a near-infrared II light source for emitting first near-infrared II light; the first near-infrared II light is near-infrared II light of one wavelength band of near-infrared II light or near-infrared II light after combining multiple different wavelength bands of near-infrared II light; a near-infrared II light scanning module for transmitting the first near-infrared II light and scanning the sample; a focusing module for focusing the first visible light on a point on the sample to excite a fluorescent substance on the sample to obtain first fluorescence; and further for focusing the first near-infrared II light on a point on the sample to excite a fluorescent substance on the sample to obtain second fluorescence; a first acquisition module for acquiring the first fluorescence; a second acquisition module for acquiring the second fluorescence. The excitation light in the present invention can be visible light of multiple different wavelength bands and near-infrared II light of multiple different wavelength bands, and can realize visible light scanning of multiple different wavelength bands and excitation light scanning of multiple different wavelength bands of near-infrared II light. Near-infrared II (NIR II: 1000nm-1700nm) imaging has characteristics such as lower autofluorescence, weaker absorption and scattering, deeper imaging depth, higher spatial resolution, and lower background noise, so that biological tissues deeper in the tissue can be imaged. The dual-channel visible light-near-infrared II light confocal microscope provided by the embodiments of the present invention can greatly broaden the fluorescence imaging coverage range, including the visible light band and near-infrared light band of 400nm-1700nm. Description of the Drawings

[0076] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0077] Figure 1 It is a structural diagram of the visible light source provided by the embodiment of the present invention;

[0078] Figure 2 It is a structural diagram of the near-infrared II light source provided by the embodiment of the present invention;

[0079] Figure 3 It is a structural diagram of the optical path provided by the embodiment of the present invention;

[0080] Figure 4Structural diagram of the first acquisition unit provided by the embodiment of the present invention;

[0081] Figure 5 Structural diagram of the second acquisition unit provided by the embodiment of the present invention;

[0082] Figure 6 Another structural diagram of the visible light source provided by the embodiment of the present invention;

[0083] Figure 7 Another structural diagram of the first acquisition unit provided by the embodiment of the present invention.

[0084] Symbol description:

[0085] 1 - First visible light source, 2 - Second visible light source, 3 - First window piece, 4 - Second window piece, 5 - First reflector, 6 - First dichroic mirror, 7 - First plano-convex lens, 8 - First optical fiber, 9 - First second near-infrared light source, 10 - Second second near-infrared light source, 11 - Third window piece, 12 - Fourth window piece, 13 - Second reflector, 14 - Third dichroic mirror, 15 - Second plano-convex lens, 16 - Second optical fiber; 17 - Visible light source optical fiber, 18 - First collimation module, 19 - Resonant galvanometer, 20 - First y-axis galvanometer-type mirror, 21 - First 4f system, 22 - First x-axis galvanometer-type mirror, 23 - Visible light scanning lens, 24 - Visible light band multi-bandpass dichroic mirror, 25 - Long-wave pass dichroic mirror, 26 - Tube lens, 27 - z-axis piezoelectric displacement stage, 28 - Objective lens, 29 - Stage, 30 - First Fresnel lens, 31 - First electric pinwheel, 32 - First multimode optical fiber, 33 - First acquisition unit, 331 - Second dichroic mirror, 332 - First filter, 333 - Second Fresnel lens, 334 - First photomultiplier tube, 335 - First signal amplifier, 336 - Second filter, 337 - Third Fresnel lens, 338 - Second photomultiplier tube, 339 - Second signal amplifier, 34 - Near-infrared light source optical fiber, 35 - Second collimation module, 36 - Near-infrared band multi-bandpass dichroic mirror, 37 - Second x-axis galvanometer-type mirror, 38 - Second 4f system, 39 - Second y-axis galvanometer-type mirror, 40 - Infrared light scanning lens, 41 - Fourth Fresnel lens, 42 - Second electric pinhole wheel, 43 - Second multimode optical fiber, 44 - Second acquisition unit, 431 - Fourth dichroic mirror, 432 - Third filter, 433 - Fifth Fresnel lens, 434 - Third photomultiplier tube, 435 - Third signal amplifier, 436 - Fourth filter, 437 - Sixth Fresnel lens, 438 - Fourth photomultiplier tube, 439 - Fourth signal amplifier, 501 - Third reflector, 502 - Fourth reflector, 503 - Fifth reflector, 504 - Sixth reflector, 505 - Seventh reflector, 506 - Eighth reflector, 21 - Third visible light source, 22 - Fourth visible light source, 41 - Fifth window piece, 42 - Sixth window piece, 61 - Fifth dichroic mirror, 62 - Sixth dichroic mirror, 45 - Seventh dichroic mirror, 46 - Eighth dichroic mirror, 47 - Fifth filter, 48 - Sixth filter, 49 - Seventh Fresnel lens, 50 - Eighth Fresnel lens, 51 - Fifth photomultiplier tube, 52 - Sixth photomultiplier tube, 53 - Fifth signal amplifier, 54 - Sixth signal amplifier. Detailed implementation mode

[0086] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0087] The terms "first", "second", "third", etc. (if any) in the specification and claims of the present invention and the above-mentioned drawings are used to distinguish similar objects and do not necessarily need to describe a specific order or sequence. It should be understood that such described objects can be interchanged under appropriate circumstances. In addition, the terms "include" and "have" and any variations thereof are intended to cover non-exclusive inclusion.

[0088] In the present invention, the following-described drawings and the embodiments used to describe the principles disclosed in the present invention are only for illustration and should not be construed as limiting the scope of the disclosure of the present invention. Those skilled in the art will understand that the principles of the present invention can be implemented in any appropriately arranged system. Exemplary embodiments will be described in detail, and examples of these embodiments are shown in the drawings. In addition, a terminal according to an exemplary embodiment will be described in detail with reference to the drawings. The same reference numerals in the drawings refer to the same elements.

[0089] The terms used in the specification of the present invention are only used to describe specific embodiments and do not intend to show the concept of the present invention. Unless there is a clearly different meaning in the context, expressions used in the singular form cover those in the plural form. In the specification of the present invention, it should be understood that terms such as "include", "have", and "contain" are intended to indicate the possibility of the existence of the features, numbers, steps, actions, or combinations thereof disclosed in the specification of the present invention, and do not intend to exclude the possibility of the existence or addition of one or more other features, numbers, steps, actions, or combinations thereof. The same reference numerals in the drawings refer to the same parts.

[0090] The purpose of the present invention is to provide a dual-channel visible light-near-infrared second-region light confocal microscope to solve the problems existing in the prior art and achieve multi-band scanning.

[0091] To make the above objects, features, and advantages of the present invention more obvious and understandable, the present invention will be further described in detail below with reference to the drawings and specific embodiments.

[0092] This embodiment discloses a dual-channel visible light-near-infrared second-region light confocal microscope, including

[0093] A visible light source for emitting first visible light. The first visible light is visible light of one band of visible light or visible light after combining multiple different bands of visible light;

[0094] A visible light scanning module for transmitting first visible light and scanning a sample at a first speed or a second speed. The first speed is higher than the second speed.

[0095] A second near-infrared light source for emitting first second near-infrared light. The first second near-infrared light is second near-infrared light of one band or second near-infrared light obtained by combining second near-infrared lights of multiple different bands.

[0096] A second near-infrared light scanning module for transmitting first second near-infrared light and scanning a sample.

[0097] A focusing module for focusing the first visible light on a point on the sample to excite a fluorescent substance on the sample to obtain first fluorescence. It is also used to focus the first second near-infrared light on a point on the sample to excite a fluorescent substance on the sample to obtain second fluorescence.

[0098] A first acquisition module for acquiring the first fluorescence.

[0099] A second acquisition module for acquiring the second fluorescence.

[0100] As Figure 1 shown, the visible light source includes a first visible light source 1, a second visible light source 2, a first window pane 3, a second window pane 4, a first mirror 5, a first dichroic mirror 6, a first plano-convex lens 7, and a first optical fiber 8. The first visible light source 1 is used to emit second visible light, and the second visible light source 2 is used to emit third visible light. The second visible light and the third visible light have different bands. The first window pane 3 and the first mirror 5 are sequentially arranged on the propagation path of the second visible light. The first window pane 3 is used to deflect the second visible light, and the first mirror 5 is used to reflect the deflected second visible light. The second window pane 4 and the first dichroic mirror 6 are sequentially arranged on the propagation path of the third visible light, and the first dichroic mirror 6 is also arranged on the propagation path of the deflected second visible light. The second window pane 4 is used to deflect the third visible light, and the first dichroic mirror 6 is used to reflect the deflected third visible light and transmit the deflected second visible light, and also combine the deflected third visible light and the deflected second visible light into a beam of light, denoted as fourth visible light. The first plano-convex lens 7 is arranged on the propagation path of the fourth visible light, and the first plano-convex lens 7 is used to focus and couple the fourth visible light into the first optical fiber 8.

[0101] As Figure 2As shown in the figure, the near-infrared second-region light source includes a first near-infrared second-region light source 9, a second near-infrared second-region light source 10, a third window piece 11, a fourth window piece 12, a second reflecting mirror 13, a third dichroic mirror 14, a second plano-convex lens 15, and a second optical fiber 16. The first near-infrared second-region light source 9 is used to emit second near-infrared second-region light, and the second near-infrared second-region light source 10 is used to emit third near-infrared second-region light. The bands of the second near-infrared second-region light and the third near-infrared second-region light are different. The third window piece 11 and the second reflecting mirror 13 are sequentially arranged on the propagation path of the second near-infrared second-region light. The third window piece 11 is used to deflect the second near-infrared second-region light, and the second reflecting mirror 13 is used to reflect the deflected second near-infrared second-region light. The fourth window piece 12 and the third dichroic mirror 14 are sequentially arranged on the propagation path of the third near-infrared second-region light. The third dichroic mirror 14 is also arranged on the propagation path of the deflected second near-infrared second-region light. The fourth window piece 12 is used to deflect the third near-infrared second-region light, and the third dichroic mirror 14 is used to reflect the deflected third near-infrared second-region light and transmit the deflected second near-infrared second-region light, and also combines the deflected third near-infrared second-region light and the deflected second near-infrared second-region light into a beam of light, denoted as the fourth near-infrared second-region light. The second plano-convex lens 15 is arranged on the propagation path of the fourth near-infrared second-region light. The second plano-convex lens 15 is used to focus the fourth near-infrared second-region light and couple it into the second optical fiber 16. Each window piece in this embodiment can be a flat glass, and the refraction of light passing through the glass is used to control the deflection of the optical path, and its thickness is determined by the actual optical path.

[0102] As Figure 3 shown, the visible light scanning module includes a visible light source optical fiber 17, a first collimation module 18, a resonant galvanometer 19, a first y-axis galvanometer mirror 20, a first 4f system 21, a first x-axis galvanometer mirror 22, a visible light scanning lens 23, and a visible light band multi-bandpass dichroic mirror 24 that are sequentially arranged on the first visible light propagation path. The visible light source optical fiber 17 is used to limit the first visible light to a point source and transmit the first visible light. The first collimation module 18 is used to collimate the first visible light. The resonant galvanometer 19 and the first y-axis galvanometer mirror 20 are used to scan the sample at a first speed. The first y-axis galvanometer mirror 20 and the first x-axis galvanometer mirror 22 are used to scan the sample at a second speed, and the first speed is higher than the second speed. The first 4f system 21 is used to collimate and optimize the optical path. The visible light scanning lens 23 is used to adapt to the wavelength of the first visible light and reduce aberration. The visible light band multi-bandpass dichroic mirror 24 is used to reflect the first visible light.

[0103] In this embodiment, the second near-infrared light scanning module includes a near-infrared light source optical fiber 34, a second collimation module 35, a near-infrared multi-bandpass dichroic mirror 36, a second x-axis galvanometric mirror 37, a second 4f system 38, a second y-axis galvanometric mirror 39, and an infrared light scanning lens 40, which are sequentially arranged on the propagation path of the first second near-infrared light. The near-infrared light source optical fiber 34 is used to confine the first second near-infrared light into a point light source and transmit the first second near-infrared light. The second collimation module 35 is used to collimate the first second near-infrared light. The near-infrared multi-bandpass dichroic mirror 36 is used to reflect the first second near-infrared light. The second x-axis galvanometric mirror 37 and the second y-axis galvanometric mirror 39 are used to scan the sample, specifically for slow scanning. The second 4f system 38 is used for collimation and optimizing the optical path. The infrared light scanning lens 40 is used to adapt to the wavelength of the first second near-infrared light and reduce aberration.

[0104] In this embodiment, the focusing module includes a long-pass dichroic mirror 25, a tube lens 26, a z-axis piezoelectric displacement stage 27, an objective lens 28, and a stage 29. The long-pass dichroic mirror 25 is arranged on the propagation path of the first visible light reflected by the visible light multi-bandpass dichroic mirror 24 and the first second near-infrared light transmitted by the infrared light scanning lens 40. The long-pass dichroic mirror 25 is used to reflect the first visible light and transmit the first second near-infrared light. The first visible light or the first second near-infrared light sequentially passes through the long-pass dichroic mirror 25, the tube lens 26, the z-axis piezoelectric displacement stage 27, and the objective lens 28 and then focuses on a point on the sample on the stage 29. The stage 29 is used to carry the sample and adjust the scanning area of the x-y plane of the sample. The z-axis piezoelectric displacement stage 27 is used to adjust the z-axis height of the objective lens 28. The objective lens 28 is used to focus the first visible light on a point on the sample, excite the fluorescent substance on the sample, and obtain the first fluorescence. The objective lens 28 is also used to focus the first second near-infrared light on a point on the sample, excite the fluorescent substance on the sample, and obtain the second fluorescence. The tube lens 26 is used to correct the phase difference of the objective lens 28.

[0105] Further, the first acquisition module includes a first Fresnel lens 30, a first motorized pinhole wheel 31, a first multimode optical fiber 32, and a first acquisition unit 33. The objective lens 28, the z-axis piezoelectric displacement stage 27, the tube lens 26, the long-pass dichroic mirror 25, and the visible light band multi-band dichroic mirror 24 are further sequentially arranged on the propagation path of the first fluorescence. The long-pass dichroic mirror 25 is further used to reflect the first fluorescence, and the visible light band band-pass dichroic mirror 24 is further used to transmit the first fluorescence. The first Fresnel lens 30 is arranged on the propagation path of the first fluorescence transmitted by the visible light band band-pass dichroic mirror 24, and the first Fresnel lens 30 is used to focus the first fluorescence on the first motorized pinhole wheel 31. The first motorized pinhole wheel 31 is used to adjust the pinhole size to reduce external fluorescence noise. One end of the first multimode optical fiber 32 is connected to the first motorized pinhole wheel 31, and the first multimode optical fiber 32 is used to transmit the first fluorescence. The first acquisition unit 33 is connected to the other end of the first multimode optical fiber 32, and the first acquisition unit 33 is used to acquire the first fluorescence. The first motorized pinhole wheel 31 in this embodiment can adjust the pinhole size in the range of 2 nm to 45 μm.

[0106] Further, the second acquisition module includes a fourth Fresnel lens 41, a second motorized pinhole wheel 42, a second multimode optical fiber 43, and a second acquisition unit 44. The objective lens 28, the z-axis piezoelectric displacement stage 27, the tube lens 26, the long-pass dichroic mirror 25, and the infrared light scanning lens 40 are further sequentially arranged on the propagation path of the second fluorescence. Both the long-pass dichroic mirror 25 and the near-infrared band multi-band dichroic mirror 36 are used to transmit the second fluorescence. The fourth Fresnel lens 41 is arranged on the propagation path of the second fluorescence transmitted by the near-infrared band multi-band dichroic mirror 36. The fourth Fresnel lens 41 is used to focus the second fluorescence on the second motorized pinhole wheel 42. The second motorized pinhole wheel 42 is used to adjust the pinhole size to reduce external fluorescence noise. One end of the second multimode optical fiber 43 is connected to the second motorized pinhole wheel 42 and is used to transmit the second fluorescence. The second acquisition unit 44 is connected to the other end of the second multimode optical fiber 43 and is used to acquire the second fluorescence. The second motorized pinhole wheel 42 in this embodiment can adjust the pinhole size in the range of 2 nm to 45 μm.

[0107] Such as Figure 4As shown, the first acquisition unit 33 includes a second dichroic mirror 331, a first filter 332, a second Fresnel lens 333, a first photomultiplier tube 334, a first signal amplifier 335, a second filter 336, a third Fresnel lens 337, a second photomultiplier tube 338, and a second signal amplifier 339. The second dichroic mirror 331 is disposed on the propagation path of the first fluorescence for splitting the first fluorescence, transmitting the first-band fluorescence in the first fluorescence and reflecting the second-band fluorescence in the first fluorescence. The first-band fluorescence and the second-band fluorescence respectively correspond to the bands of the visible light emitted by the visible light source, and can respectively correspond to the second visible light and the third visible light, so as to achieve multi-channel fluorescence imaging. The first filter 332, the second Fresnel lens 333, and the first photomultiplier tube 334 are sequentially disposed on the propagation path of the first-band fluorescence transmitted by the second dichroic mirror 331. The first filter 332 is used for filtering the first-band fluorescence, the second Fresnel lens 333 is used for focusing the first-band fluorescence into the first photomultiplier tube 334, and the first photomultiplier tube 334 is used for converting the first-band fluorescence focused by the second Fresnel lens 333 into a first electrical signal. One end of the first signal amplifier 335 is connected to the first photomultiplier tube 334, and the first signal amplifier 335 is used for amplifying the first electrical signal. The second filter 336, the third Fresnel lens 337, and the second photomultiplier tube 338 are sequentially disposed on the propagation path of the second-band fluorescence reflected by the second dichroic mirror 331. The second filter 336 is used for filtering the second-band fluorescence, the third Fresnel lens 337 is used for focusing the second-band fluorescence into the second photomultiplier tube 338, and the second photomultiplier tube 338 is used for converting the second-band fluorescence focused by the third Fresnel lens 337 into a second electrical signal. One end of the second signal amplifier 339 is connected to the second photomultiplier tube 338, and the second signal amplifier 339 is used for amplifying the second electrical signal. The other end of the first signal amplifier 335 is connected to the other end of the second signal amplifier 339, and the other end of the first signal amplifier 335 or the other end of the second signal amplifier 339 is further connected to an external processor.

[0108] As Figure 5As shown, the second acquisition unit 44 includes a fourth dichroic mirror 431, a third filter 432, a fifth Fresnel lens 433, a third photomultiplier tube 434, a third signal amplifier 435, a fourth filter 436, a sixth Fresnel lens 437, a fourth photomultiplier tube 438, and a fourth signal amplifier 439. The fourth dichroic mirror 431 is disposed on the propagation path of the second fluorescence, and is used for splitting the second fluorescence, transmitting the third-band fluorescence in the second fluorescence and reflecting the fourth-band fluorescence in the second fluorescence. The third-band fluorescence and the fourth-band fluorescence respectively correspond to the bands of the second near-infrared light emitted by the near-infrared light source in the second near-infrared region, and can respectively correspond to the second near-infrared light in the second near-infrared region and the third near-infrared light in the second near-infrared region, so as to achieve multi-channel fluorescence imaging. The third filter 432, the fifth Fresnel lens 433, and the third photomultiplier tube 434 are sequentially disposed on the propagation path of the third-band fluorescence transmitted by the fourth dichroic mirror 431. The third filter 432 is used for filtering the third-band fluorescence, the fifth Fresnel lens 433 is used for focusing the third-band fluorescence into the third photomultiplier tube 434, and the third photomultiplier tube 434 is used for converting the third-band fluorescence focused by the fifth Fresnel lens 433 into a third electrical signal. One end of the third signal amplifier 435 is connected to the third photomultiplier tube 434, and the third signal amplifier 435 is used for amplifying the third electrical signal. The fourth filter 436, the sixth Fresnel lens 437, and the fourth photomultiplier tube 438 are sequentially disposed on the propagation path of the fourth-band fluorescence reflected by the fourth dichroic mirror 431. The fourth filter 436 is used for filtering the fourth-band fluorescence, the sixth Fresnel lens 437 is used for focusing the fourth-band fluorescence into the fourth photomultiplier tube 438, and the fourth photomultiplier tube 438 is used for converting the fourth-band fluorescence focused by the sixth Fresnel lens 437 into a fourth electrical signal. One end of the fourth signal amplifier 439 is connected to the fourth photomultiplier tube 438, and the fourth signal amplifier 439 is used for amplifying the fourth electrical signal. The other end of the third signal amplifier 435 is connected to the other end of the fourth signal amplifier 439, and the other end of the third signal amplifier 435 or the other end of the fourth signal amplifier 439 is also connected to an external processor.

[0109] The usage process of the present invention is as follows:

[0110] As Figure 3As shown, the sample coated with a fluorescent substance is placed on the stage 29. The visible light source or the second near-infrared light source is turned on. The visible light source can emit visible light in multiple bands. The visible light passes through the visible light source optical fiber 17, the first collimation module 18, the third mirror 501, the fourth mirror 502, the resonant galvanometer 19, the first y-axis galvanometer mirror 20, the first 4f system 21, the first x-axis galvanometer mirror 22, the fifth mirror 503, the visible light scanning lens 23, the sixth mirror 504, the visible light band multi-bandpass dichroic mirror 24, the long-pass dichroic mirror 25, the tube lens 26, the z-axis piezoelectric displacement stage 27, and the objective lens 28 in sequence, and then irradiates the sample on the stage 29. The fluorescent substance on the sample is excited, and the first fluorescence after excitation passes through the objective lens 28, the z-axis piezoelectric displacement stage 27, the tube lens 26, the long-pass dichroic mirror 25, the visible light band multi-bandpass dichroic mirror 24, the first Fresnel lens 30, the first motorized pinwheel 31, and the first multimode optical fiber 32 in sequence and is collected by the first acquisition unit 33. Among them, the visible light band multi-bandpass dichroic mirror 24 can both reflect visible light and transmit the first fluorescence.

[0111] The resonant galvanometer 19 and the first y-axis galvanometer mirror 20 form a fast scanning galvanometer pair for the visible light optical path, which can perform fast laser scanning on the sample to achieve real-time observation effects. The first y-axis galvanometer mirror 20 and the first x-axis galvanometer mirror 22 form a slow scanning galvanometer pair for the visible light optical path, which can perform slow laser scanning on the sample, improve the image resolution while ensuring a certain imaging rate, and display more details of the sample.

[0112] The near-infrared second-region light source can emit near-infrared second-region light in multiple bands. The near-infrared second-region light sequentially passes through the near-infrared light source optical fiber 34, the second collimation module 35, the near-infrared multi-bandpass dichroic mirror 36, the seventh reflector 505, the second x-axis galvanometric scanning mirror 37, the second 4f system 38, the second y-axis galvanometric scanning mirror 39, the infrared light scanning lens 40, the eighth reflector 506, the long-pass dichroic mirror 25, the tube lens 26, the z-axis piezoelectric displacement stage 27, and the objective lens 28, and then irradiates the sample on the stage 29. The fluorescent substance on the sample is excited, and the excited second fluorescence sequentially passes through the objective lens 28, the z-axis piezoelectric displacement stage 27, the tube lens 26, the long-pass dichroic mirror 25, the eighth reflector 506, the infrared light scanning lens 40, the second y-axis galvanometric scanning mirror 39, the second 4f system 38, the second x-axis galvanometric scanning mirror 37, the seventh reflector 505, the near-infrared multi-bandpass dichroic mirror 36, the fourth Fresnel lens 41, the second motorized pinhole wheel 42, and the second multimode optical fiber 43, and is then collected by the second acquisition unit 44. Among them, the long-pass dichroic mirror 25 can reflect visible light and also transmit near-infrared second-region light, enabling visible light and near-infrared second-region light to enter the tube lens 26 together. The near-infrared multi-bandpass dichroic mirror 36 can both reflect near-infrared second-region light and transmit the second fluorescence.

[0113] The second x-axis galvanometric scanning mirror 37 and the second y-axis galvanometric scanning mirror 39 form a slow-scanning galvanometric mirror pair for the near-infrared second-region light optical path, which can perform slow laser scanning on the sample, improve the image resolution while ensuring a certain imaging rate, and display more details of the sample.

[0114] In another embodiment, both the visible light source and the near-infrared second-region light source include four light sources with different bands, and both the first acquisition unit and the second acquisition unit include four photomultiplier tubes and their corresponding components. Specifically as follows:

[0115] Such as Figure 6As shown in the figure, the visible light source further includes a third visible light source 21, a fourth visible light source 22, a fifth window 41, a sixth window 42, a fifth dichroic mirror 61, and a sixth dichroic mirror 62. The light emitted by the third visible light source 21 enters the fifth dichroic mirror 61 through the fifth window 41. The fifth dichroic mirror 61 transmits the visible light emitted by the first visible light source 1 and the second visible light source 2, and reflects the visible light emitted by the third visible light source 21. The light emitted by the fourth visible light source 22 enters the sixth dichroic mirror 62 through the sixth window 42. The sixth dichroic mirror 62 transmits the visible light emitted by the first visible light source 1, the second visible light source 2, and the third visible light source 21, and reflects the visible light emitted by the fourth visible light source 22. The sixth dichroic mirror 62 also combines the visible light emitted by the first visible light source 1, the second visible light source 2, the third visible light source 21, and the fourth visible light source 22 into a single beam of light. The same applies to the second near-infrared light source, which will not be elaborated here.

[0116] As Figure 7 shown in the figure, the first acquisition unit further includes a seventh dichroic mirror 45, an eighth dichroic mirror 46, a fifth filter 47, a sixth filter 48, a seventh Fresnel lens 49, an eighth Fresnel lens 50, a fifth photomultiplier tube 51, a sixth photomultiplier tube 52, a fifth signal amplifier 53, and a sixth signal amplifier 54. The first fluorescence is split by the eighth dichroic mirror 46 and then collected by the first photomultiplier tube 334, the second photomultiplier tube 338, the fifth photomultiplier tube 51, and the sixth photomultiplier tube 52 respectively. The same applies to the second acquisition unit, which will not be elaborated here.

[0117] According to the specific embodiments provided by the present invention, the present invention discloses the following technical effects:

[0118] The excitation light in the present invention can be visible light of multiple different wavelength bands and second near-infrared light of multiple different wavelength bands, enabling visible light scanning of multiple different wavelength bands and second near-infrared excitation light scanning of multiple different wavelength bands, thereby greatly expanding the fluorescence imaging coverage range to 400nm - 1700nm. Second near-infrared (NIR II: 1000nm - 1700nm) imaging has characteristics such as lower autofluorescence, weaker absorption and scattering, resulting in deeper imaging depth, higher spatial resolution, and lower background noise, thus enabling imaging of biological tissues deeper in the tissue.

[0119] The resonant galvanometer and the first y-axis galvanometer in the present invention form a fast scanning galvanometer pair for the visible light optical path, which can perform fast visible light laser scanning on the sample to achieve real-time observation effects.

[0120] In this specification, the various embodiments are described in a progressive manner. Each embodiment focuses on the differences from other embodiments. For the same or similar parts among the various embodiments, reference can be made to each other.

[0121] In this article, specific examples are used to elaborate on the principles and implementation manners of the present invention. The descriptions of the above embodiments are only used to help understand the core idea of the present invention. At the same time, for those of ordinary skill in the art, according to the idea of the present invention, there will be changes in the specific implementation manners and application scopes. In summary, the content of this specification should not be construed as a limitation on the present invention.

Claims

1. A dual-channel visible light-near-infrared second near-infrared window light confocal microscope, characterized in that, Including: A visible light source for emitting a first visible light; the first visible light is visible light of one wavelength band or visible light obtained by combining visible lights of multiple different wavelength bands; A visible light scanning module for transmitting the first visible light and scanning a sample at a first speed or a second speed; the first speed is higher than the second speed, and the visible light scanning module includes a visible light source optical fiber, a first collimation module, a resonant galvanometer, a first y-axis galvanometer mirror, a first 4f system, a first x-axis galvanometer mirror, a visible light scanning lens, and a visible light band multi-bandpass dichroic mirror sequentially arranged on the propagation path of the first visible light, where: The visible light source optical fiber is used to confine the first visible light to a point light source and transmit the first visible light; The first collimation module is used to collimate the first visible light; The resonant galvanometer and the first y-axis galvanometer mirror are used to scan the sample at the first speed; The first y-axis galvanometer mirror and the first x-axis galvanometer mirror are used to scan the sample at the second speed, and the first speed is higher than the second speed; The first 4f system is used to collimate and optimize the optical path; The visible light scanning lens is used to adapt to the wavelength of the first visible light and reduce aberration; The visible light band multi-bandpass dichroic mirror is used to reflect the first visible light; A near-infrared second-region light source for emitting a first near-infrared second-region light; the first near-infrared second-region light is near-infrared second-region light of one wavelength band or near-infrared second-region light obtained by combining near-infrared second-region lights of multiple different wavelength bands; A near-infrared second-region light scanning module for transmitting the first near-infrared second-region light and scanning the sample, and the near-infrared second-region light scanning module includes a near-infrared light source optical fiber, a second collimation module, a near-infrared band multi-bandpass dichroic mirror, a second x-axis galvanometer mirror, a second 4f system, a second y-axis galvanometer mirror, and an infrared light scanning lens sequentially arranged on the propagation path of the first near-infrared second-region light, where: The near-infrared light source optical fiber is used to confine the first near-infrared second-region light to a point light source and transmit the first near-infrared second-region light; The second collimation module is used to collimate the first near-infrared second-region light; The near-infrared band multi-bandpass dichroic mirror is used to reflect the first near-infrared second-region light; The second x-axis galvanometer mirror and the second y-axis galvanometer mirror are used to scan the sample; The second 4f system is used to collimate and optimize the optical path; The infrared light scanning lens is used to adapt to the wavelength of the first near-infrared second-region light and reduce aberration; A focusing module for focusing the first visible light on a point on the sample to excite a fluorescent substance on the sample to obtain a first fluorescence; and also for focusing the first near-infrared second-region light on a point on the sample to excite a fluorescent substance on the sample to obtain a second fluorescence, and the focusing module includes a long-pass dichroic mirror, a tube lens, a z-axis piezoelectric displacement stage, an objective lens, and a stage, where: The long-pass dichroic mirror is disposed on the propagation paths of the first visible light reflected by the visible light band multi-band dichroic mirror and the first second near-infrared light transmitted by the infrared light scanning lens; the long-pass dichroic mirror is configured to reflect the first visible light and transmit the first second near-infrared light; The first visible light or the first second near-infrared light sequentially passes through the long-pass dichroic mirror, the tube lens, the z-axis piezoelectric displacement stage, and the objective lens and then focuses on a point on the sample on the stage; The stage is used to carry the sample and adjust the scanning area of the x-y plane of the sample; The z-axis piezoelectric displacement stage is used to adjust the z-axis height of the objective lens; The objective lens is used to focus the first visible light on a point on the sample to excite the fluorescent substance on the sample to obtain the first fluorescence; the objective lens is also used to focus the first second near-infrared light on a point on the sample to excite the fluorescent substance on the sample to obtain the second fluorescence; The tube lens is used to correct the phase difference of the objective lens; A first acquisition module, configured to acquire the first fluorescence, the first acquisition module includes a first Fresnel lens, a first motorized pinhole wheel, a first multimode optical fiber, and a first acquisition unit, wherein: The objective lens, the z-axis piezoelectric displacement stage, the tube lens, the long-pass dichroic mirror, and the visible light band multi-band dichroic mirror are also sequentially disposed on the propagation path of the first fluorescence; the long-pass dichroic mirror is also used to reflect the first fluorescence; the visible light band band-pass dichroic mirror is also used to transmit the first fluorescence; The first Fresnel lens is disposed on the propagation path of the first fluorescence transmitted by the visible light band band-pass dichroic mirror; the first Fresnel lens is used to focus the first fluorescence on the first motorized pinhole wheel; The first motorized pinhole wheel is used to adjust the pinhole size to reduce external fluorescence noise; One end of the first multimode optical fiber is connected to the first motorized pinhole wheel; the first multimode optical fiber is used to transmit the first fluorescence; The first acquisition unit is connected to the other end of the first multimode optical fiber; the first acquisition unit is used to acquire the first fluorescence; A second acquisition module, configured to acquire the second fluorescence, the second acquisition module includes a fourth Fresnel lens, a second motorized pinhole wheel, a second multimode optical fiber, and a second acquisition unit, wherein: The objective lens, the z-axis piezoelectric displacement stage, the tube lens, the long-pass dichroic mirror, and the infrared light scanning lens are also sequentially disposed on the propagation path of the second fluorescence; the long-pass dichroic mirror and the near-infrared band multi-band dichroic mirror are both used to transmit the second fluorescence; The fourth Fresnel lens is disposed on the propagation path of the second fluorescence transmitted by the near-infrared band multi-band dichroic mirror; the fourth Fresnel lens is used to focus the second fluorescence on the second motorized pinhole wheel; The second motorized pinhole wheel is used to adjust the pinhole size to reduce external fluorescence noise; One end of the second multimode optical fiber is connected to the second motorized pinhole wheel; the second multimode optical fiber is used to transmit the second fluorescence; The second acquisition unit is connected to the other end of the second multimode optical fiber; the second acquisition unit is used to acquire the second fluorescence.

2. The dual-channel visible light-near-infrared second window light confocal microscope according to claim 1, wherein The visible light source includes a first visible light source, a second visible light source, a first window plate, a second window plate, a first reflector, a first dichroic mirror, a first plano-convex lens, and a first optical fiber; The first visible light source is used to emit second visible light; the second visible light source is used to emit third visible light; the wavelength bands of the second visible light and the third visible light are different; The first window plate and the first reflector are sequentially arranged on the propagation path of the second visible light; the first window plate is used to deflect the second visible light; the first reflector is used to reflect the deflected second visible light; The second window plate and the first dichroic mirror are sequentially arranged on the propagation path of the third visible light; the first dichroic mirror is also arranged on the propagation path of the deflected second visible light; the second window plate is used to deflect the third visible light; the first dichroic mirror is used to reflect the deflected third visible light and transmit the deflected second visible light, and also combines the deflected third visible light and the deflected second visible light into a beam of light, denoted as fourth visible light; The first plano-convex lens is arranged on the propagation path of the fourth visible light; the first plano-convex lens is used to focus and couple the fourth visible light into the first optical fiber.

3. The dual-channel visible light-near-infrared II region light confocal microscope according to claim 1, characterized in that, The near-infrared second-region light source includes a first near-infrared second-region light source, a second near-infrared second-region light source, a third window plate, a fourth window plate, a second reflector, a third dichroic mirror, a second plano-convex lens, and a second optical fiber; The first near-infrared second-region light source is used to emit second near-infrared second-region light; the second near-infrared second-region light source is used to emit third near-infrared second-region light; the wavelength bands of the second near-infrared second-region light and the third near-infrared second-region light are different; The third window plate and the second reflector are sequentially arranged on the propagation path of the second near-infrared second-region light; the third window plate is used to deflect the second near-infrared second-region light; the second reflector is used to reflect the deflected second near-infrared second-region light; The fourth window plate and the third dichroic mirror are sequentially arranged on the propagation path of the third near-infrared second-region light; the third dichroic mirror is also arranged on the propagation path of the deflected second near-infrared second-region light; the fourth window plate is used to deflect the third near-infrared second-region light; the third dichroic mirror is used to reflect the deflected third near-infrared second-region light and transmit the deflected second near-infrared second-region light, and also combines the deflected third near-infrared second-region light and the deflected second near-infrared second-region light into a beam of light, denoted as fourth near-infrared second-region light; The second plano-convex lens is arranged on the propagation path of the fourth near-infrared second-region light; the second plano-convex lens is used to focus and couple the fourth near-infrared second-region light into the second optical fiber.

4. The dual-path visible light-near-infrared second-region light confocal microscope according to claim 1, wherein The first acquisition unit includes a second dichroic mirror, a first filter, a second Fresnel lens, a first photomultiplier tube, a first signal amplifier, a second filter, a third Fresnel lens, a second photomultiplier tube, and a second signal amplifier; The second dichroic mirror is disposed on the propagation path of the first fluorescence; the second dichroic mirror is configured to split the first fluorescence, transmit the first-band fluorescence in the first fluorescence, and reflect the second-band fluorescence in the first fluorescence; the first-band fluorescence and the second-band fluorescence respectively correspond to the bands of the visible light emitted by the visible light source; The first filter, the second Fresnel lens, and the first photomultiplier tube are sequentially disposed on the propagation path of the first-band fluorescence transmitted by the second dichroic mirror; the first filter is configured to filter the first-band fluorescence; the second Fresnel lens is configured to focus the first-band fluorescence into the first photomultiplier tube; The first photomultiplier tube is configured to convert the first-band fluorescence focused by the second Fresnel lens into a first electrical signal; One end of the first signal amplifier is connected to the first photomultiplier tube, and the first signal amplifier is configured to amplify the first electrical signal; The second filter, the third Fresnel lens, and the second photomultiplier tube are sequentially disposed on the propagation path of the second-band fluorescence reflected by the second dichroic mirror; the second filter is configured to filter the second-band fluorescence; the third Fresnel lens is configured to focus the second-band fluorescence into the second photomultiplier tube; The second photomultiplier tube is configured to convert the second-band fluorescence focused by the third Fresnel lens into a second electrical signal; One end of the second signal amplifier is connected to the second photomultiplier tube, and the second signal amplifier is configured to amplify the second electrical signal; The other end of the first signal amplifier is connected to the other end of the second signal amplifier; the other end of the first signal amplifier or the other end of the second signal amplifier is further connected to an external processor.

5. The dual-channel visible light-near-infrared second window light confocal microscope according to claim 1, wherein, The second acquisition unit includes a fourth dichroic mirror, a third filter, a fifth Fresnel lens, a third photomultiplier tube, a third signal amplifier, a fourth filter, a sixth Fresnel lens, a fourth photomultiplier tube, and a fourth signal amplifier; The fourth dichroic mirror is disposed on the propagation path of the second fluorescence; the fourth dichroic mirror is configured to split the second fluorescence, transmit the third-band fluorescence in the second fluorescence, and reflect the fourth-band fluorescence in the second fluorescence; the third-band fluorescence and the fourth-band fluorescence respectively correspond to the bands of the second near-infrared light emitted by the second near-infrared light source; The third filter, the fifth Fresnel lens, and the third photomultiplier tube are sequentially disposed on the propagation path of the third-band fluorescence transmitted by the fourth dichroic mirror; the third filter is configured to filter the third-band fluorescence; the fifth Fresnel lens is configured to focus the third-band fluorescence into the third photomultiplier tube; The third photomultiplier tube is configured to convert the third-band fluorescence focused by the fifth Fresnel lens into a third electrical signal; One end of the third signal amplifier is connected to the third photomultiplier tube, and the third signal amplifier is configured to amplify the third electrical signal; The fourth filter, the sixth Fresnel lens, and the fourth photomultiplier tube are sequentially arranged on the propagation path of the fourth-band fluorescence reflected by the fourth dichroic mirror; the fourth filter is used to filter the fourth-band fluorescence; the sixth Fresnel lens is used to focus the fourth-band fluorescence into the fourth photomultiplier tube; The fourth photomultiplier tube is used to convert the fourth-band fluorescence focused by the sixth Fresnel lens into a fourth electrical signal; One end of the fourth signal amplifier is connected to the fourth photomultiplier tube, and the fourth signal amplifier is used to amplify the fourth electrical signal; The other end of the third signal amplifier is connected to the other end of the fourth signal amplifier; the other end of the third signal amplifier or the other end of the fourth signal amplifier is also connected to an external processor.

Images