Multiphoton imaging system, multiphoton stimulation system and multiphoton imaging and stimulation system

Abstract

The present invention relates to a multiphoton imaging system, a multiphoton stimulation system and a multiphoton imaging and stimulation system. The system switches the system optical path to a multiphoton imaging optical path, an adaptive optical multiphoton imaging optical path, a multiphoton optical stimulation optical path or a multiphoton holographic optical stimulation optical path by adjusting the position of an electrically controlled movable reflector. Compared with the prior art, the present invention can achieve high-precision multiphoton imaging and single-cell precision multiphoton optical stimulation.

Description

Technical Field

[0001] The present invention relates to the field of optical microscopic imaging, and in particular to a multiphoton imaging system, a multiphoton stimulation system and a multiphoton imaging and stimulation system. Background Art

[0002] Multi-photon microscopy (MPM), including two-photon microscopy and three-photon microscopy, is a nonlinear microscopic imaging technology that uses high-energy infrared lasers to excite fluorescent molecules to obtain high-resolution imaging of deep tissues. Compared with single-photon microscopy, multi-photon microscopy has lower excitation photon energy, which can reduce tissue damage and increase imaging depth.

[0003] Multiphoton imaging technology is widely used in fields such as neuroscience, developmental biology, and immunology, and is an indispensable tool in modern biomedical research. Although two-photon microscopy has a high imaging depth and low tissue damage, it still faces some challenges when imaging deep tissues. When imaging deep tissues, ordinary two-photon microscopes are affected by tissue heterogeneity and optical aberrations, and the imaging quality is significantly reduced. Aberrations can cause image blur and reduced resolution, especially in high-resolution imaging. Due to changes in the optical properties of tissues, the excitation spot may be out of focus after entering deep tissues, resulting in weakened fluorescence signals and reduced imaging clarity. As the excitation light penetrates deeper into the tissue, the scattering and absorption of light increase, resulting in weakened signals, increased background noise, and ultimately a decrease in the signal-to-noise ratio, making it difficult to obtain clear images in deep tissues.

[0004] Biomedical research requires not only the observation of cells, but also the manipulation of specific cells. Chemical genetics and optogenetics are commonly used to manipulate specific cells, among which optogenetics has higher temporal and spatial resolution. In addition, during two-photon imaging, single-photon optogenetics and two-photon optogenetics can be used for more precise stimulation. Specific cells can be stimulated while being observed, and the effects of activation or inhibition of specific cells on the activity of surrounding cells and on specific behaviors can be observed. Traditional two-photon optogenetics requires stimulation of target neurons one by one, which means that it cannot accurately activate multiple target neurons or regions at the same time, which is very limited for studying complex neural circuits or large-scale neural activities. Summary of the invention

[0005] The purpose of the present invention is to overcome the defects of the above-mentioned prior art and to provide a mode-switchable multi-photon imaging system, a multi-photon stimulation system and a multi-photon imaging and stimulation system. The corresponding imaging mode can be selected according to the actual application situation, and the corresponding imaging or stimulation mode can be selected according to the actual application situation to achieve high-resolution multi-photon imaging or holographic light stimulation.

[0006] The purpose of the present invention can be achieved by the following technical solutions:

[0007] A mode-switchable multi-photon imaging system comprises an imaging laser, a first electrically-controlled movable reflector, a second electrically-controlled movable reflector, a fourth electrically-controlled movable reflector, an imaging resonant galvanometer, an imaging non-resonant galvanometer, a first dichroic mirror, an objective lens, a wavefront control device, a lens and a plurality of lens groups;

[0008] The system switches the system optical path to a multiphoton imaging optical path or an adaptive optical multiphoton imaging optical path by adjusting the position of the electrically controlled movable reflector, specifically comprising:

[0009] Multiphoton imaging optical path: the first electrically-controlled movable reflector in the first position and the second electrically-controlled movable reflector in the first position are not connected to the multiphoton imaging optical path, the output of the imaging laser acts on the imaging resonant galvanometer, the output of the imaging resonant galvanometer acts on the imaging non-resonant galvanometer, the output of the imaging non-resonant galvanometer is reflected by the first dichroic mirror and acts on the objective lens through the first optical path containing the lens;

[0010] Adaptive optical multiphoton imaging optical path: the output of the imaging laser is reflected and directed by the first electrically-controlled movable mirror in the second position, and then is controlled by the wavefront control device after passing through the fourth electrically-controlled movable mirror in the first position. The output of the wavefront control device is reflected and adjusted through the second electrically-controlled movable mirror in the second position to act on the imaging resonant galvanometer. The output of the imaging resonant galvanometer acts on the imaging non-resonant galvanometer. The output of the imaging non-resonant galvanometer is reflected by the first dichroic mirror and acts on the objective lens through the first optical path containing the lens.

[0011] Preferably, the multiphoton imaging optical path and the adaptive optics multiphoton imaging optical path share the same optical path after acting on the imaging resonant galvanometer.

[0012] A mode-switchable multi-photon stimulation system, the system comprising a stimulation laser, a third electrically-controlled movable reflector, a fourth electrically-controlled movable reflector, a fifth electrically-controlled movable reflector, a sixth electrically-controlled movable reflector, a stimulation non-resonant galvanometer, a first dichroic mirror, an objective lens, a wavefront control device, a lens, and a plurality of lens groups;

[0013] The system switches the system optical path to a multi-photon light stimulation optical path or a multi-photon holographic light stimulation optical path by adjusting the position of the electrically controlled movable reflector, specifically:

[0014] Multi-photon light stimulation optical path: the output of the stimulation imaging laser is reflected and directed by the third electrically-controlled movable reflector in the first position, and then acts on the stimulation non-resonant galvanometer after being reflected and directed by the sixth electrically-controlled movable reflector in the first position. The output of the stimulation non-resonant galvanometer is transmitted through the first dichroic mirror and then acts on the objective lens through the first optical path containing the lens;

[0015] Multiphoton holographic light stimulation optical path: the third electrically-controlled movable reflector in the second position, the fourth electrically-controlled movable reflector in the second position and the sixth electrically-controlled movable reflector in the second position are not connected to the multiphoton holographic light stimulation optical path, the output of the stimulation imaging laser is expanded and then acts on the wavefront control device, the output of the wavefront control device is reflected and adjusted by the fifth electrically-controlled movable reflector in the second position and then enters the stimulation non-resonant galvanometer, the output of the stimulation non-resonant galvanometer is transmitted through the first dichroic mirror and then acts on the objective lens through the first optical path containing the lens.

[0016] Preferably, the multi-photon optical stimulation optical path and the multi-photon holographic optical stimulation optical path share the same optical path after acting on the non-resonant galvanometer to stimulate the non-resonant galvanometer.

[0017] Preferably, in the multi-photon holographic optical stimulation light path, after the direction is adjusted by the fifth electrically-controlled movable reflector in the second position, before entering the non-resonant galvanometer, it also includes: the light path lens reflected by the fifth electrically-controlled movable reflector focuses, and the zero-order light is blocked by the zero-order baffle at the focus.

[0018] Preferably, a plurality of groups of lenses for adjusting the diameter of the light beam are arranged on the optical path.

[0019] Preferably, a scanning lens, a cylindrical lens and a second dichroic mirror are sequentially arranged on the first optical path containing a lens, and the output of the cylindrical lens is reflected by the second dichroic mirror and then acts on the objective lens.

[0020] Preferably, the system further comprises a PMT, and the signal collected by the objective lens is transmitted through the second dichroic mirror, and then separated into a desired wavelength signal through a third dichroic mirror before acting on the PMT.

[0021] A mode-switchable multiphoton imaging and stimulation system comprises an imaging subsystem and a stimulation subsystem; the imaging subsystem adopts the multiphoton imaging system, and the stimulation subsystem adopts the multiphoton stimulation system.

[0022] Preferably, the multiphoton imaging optical path and the multiphoton stimulation system share the same optical path after passing through the first dichroic mirror.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] (1) By adjusting the position of the electrically controlled movable reflector, the system optical path can be switched to a multiphoton imaging optical path or an adaptive optics multiphoton imaging optical path. The corresponding imaging mode can be selected according to the actual application to achieve high-resolution multiphoton imaging. The use of multiple groups of lenses in the multiphoton imaging optical path improves the imaging resolution and wavefront control effect. The use of a wavefront controller in the adaptive optics multiphoton imaging optical path can achieve precise control of the light beam, thereby improving the imaging quality and imaging depth.

[0025] (2) The back-end optical paths of the multiphoton imaging optical path and the adaptive optics multiphoton imaging optical path are combined and shared. High-resolution multiphoton imaging in both imaging modes can be achieved as long as the spot size entering the shared segment is consistent. The combined and shared back-end optical paths also further reduce the number of optical path components in the system, further reducing costs while meeting multi-functional imaging requirements.

[0026] (3) By adjusting the position of the electrically controlled movable reflector, the system optical path can be directly switched to a multi-photon light stimulation path or a multi-photon holographic light stimulation path. The corresponding stimulation mode can be selected according to the actual application, taking into account the diversity and flexibility of the experiment.

[0027] (4) In the multi-photon holographic light stimulation optical path, a zero-order baffle is set to block the zero laser, and then the light spot is adjusted to a set size through lens expansion to fill the non-resonant galvanometer as much as possible. Then the optical path is merged with ordinary multi-photon light stimulation. Through multiple beam expansions, the light spot is magnified to a set multiple before entering the objective lens, so that high-precision multi-photon holographic light stimulation can be performed. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Schematic diagram of the optical path of the mode-switchable multi-photon imaging system in the embodiment;

[0029] Figure 2 Schematic diagram of the optical path of the mode-switchable multi-photon stimulation system in the embodiment;

[0030] Figure 3 Schematic diagram of the optical path of the mode-switchable multiphoton imaging and stimulation system in the embodiment.

[0031] Reference numerals:

[0032] 1-electrically controlled movable reflector, 2-lens group, 3-lens group, 4-electrically controlled movable reflector, 5-lens, 6-imaging resonant galvanometer, 7-lens, 8-lens, 9-stimulating non-resonant galvanometer, 10-imaging non-resonant galvanometer, 11-first dichroic mirror, 12-scanning lens, 13-reflector, 14-cylindrical lens, 15-second dichroic mirror, 16-electrically controlled movable reflector, 17-electrically controlled movable reflector, 18-lens group, 19-electrically controlled movable reflector, 20-lens group, 21-wavefront control device, 22-lens, 23-electrically controlled movable reflector, 24-lens, 25-zero-order baffle, 26-electrically controlled movable reflector, 27-lens, 28-PMT, 29-lens, 30-third dichroic mirror, 31-lens. DETAILED DESCRIPTION

[0033] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0034] Example

[0035] like Figure 1 As shown, this embodiment provides a mode-switchable multi-photon imaging system, including an imaging laser, a first electrically-controlled movable reflector 1, a second electrically-controlled movable reflector 4, a fourth electrically-controlled movable reflector 19, an imaging resonant galvanometer 6, an imaging non-resonant galvanometer 10, a first dichroic mirror 11, an objective lens 16, a wavefront control device 21, a lens, and a plurality of lens groups;

[0036] The system switches the system optical path to a multiphoton imaging optical path or an adaptive optical multiphoton imaging optical path by adjusting the position of the electrically controlled movable reflector, specifically including:

[0037] Multiphoton imaging optical path: the first electrically-controlled movable reflector 1 in the first position and the second electrically-controlled movable reflector 4 in the first position are not connected to the multiphoton imaging optical path, the output of the imaging laser acts on the imaging resonant galvanometer 6, the output of the imaging resonant galvanometer 6 acts on the imaging non-resonant galvanometer 10, the output of the imaging non-resonant galvanometer 10 is reflected by the first dichroic mirror 11 and acts on the objective lens 16 through the first optical path containing the lens;

[0038] Adaptive optical multiphoton imaging optical path: the output of the imaging laser is reflected and directed by the first electrically-controlled movable mirror 1 in the second position, then by the fourth electrically-controlled movable mirror 19 in the first position, and then controlled by the wavefront control device 21. The output of the wavefront control device 21 is reflected and adjusted through the second electrically-controlled movable mirror 4 in the second position to act on the imaging resonant galvanometer 6. The output of the imaging resonant galvanometer 6 acts on the imaging non-resonant galvanometer 10. The output of the imaging non-resonant galvanometer 10 is reflected by the first dichroic mirror 11 and acts on the objective lens 16 through the first optical path containing the lens.

[0039] like Figure 2 As shown, this embodiment provides a mode-switchable multi-photon stimulation system, including a stimulation laser, a third electrically-controlled movable reflector 17, a fourth electrically-controlled movable reflector 19, a fifth electrically-controlled movable reflector 23, a sixth electrically-controlled movable reflector 26, a stimulation non-resonant galvanometer 9, a first dichroic mirror 11, an objective lens 16, a wavefront control device 21, a lens, and a plurality of lens groups;

[0040] The system switches the system optical path to a multi-photon light stimulation optical path or a multi-photon holographic light stimulation optical path by adjusting the position of the electrically controlled movable reflector, specifically:

[0041] Multi-photon light stimulation optical path: the output of the stimulation imaging laser is reflected and adjusted in direction by the third electrically-controlled movable reflector 17 in the first position, and then acts on the stimulation non-resonant galvanometer 9 after being reflected and adjusted in direction by the sixth electrically-controlled movable reflector 26 in the first position. The output of the stimulation non-resonant galvanometer 9 is transmitted through the first dichroic mirror 11 and then acts on the objective lens 16 through the first optical path containing the lens;

[0042] Multi-photon holographic light stimulation optical path: the third electrically-controlled movable reflector 17 in the second position, the fourth electrically-controlled movable reflector 19 in the second position, and the sixth electrically-controlled movable reflector 26 in the second position are not connected to the multi-photon holographic light stimulation optical path, the output of the stimulation imaging laser is expanded and acts on the wavefront control device 21, the output of the wavefront control device 21 is reflected and adjusted by the fifth electrically-controlled movable reflector 23 in the second position and then enters the stimulation non-resonant galvanometer 9, the output of the stimulation non-resonant galvanometer 9 is transmitted through the first dichroic mirror 11 and acts on the objective lens 16 through the first optical path containing the lens.

[0043] In neuroscience research, we first use multiphoton imaging to image the morphology and distribution of neurons, and then switch to multiphoton stimulation to stimulate specific neurons to observe their reactions and signal transmission processes. Implementing both imaging and stimulation in the same optical system greatly improves experimental efficiency and accuracy. Figure 3As shown, this embodiment provides a mode-switchable multi-photon imaging and stimulation system, including an imaging laser (Laser IMG), a stimulation laser (Laser STI), a first electrically-controlled movable reflector 1, a second electrically-controlled movable reflector 4, a fourth electrically-controlled movable reflector 19, a fifth electrically-controlled movable reflector 23, an imaging resonant galvanometer 6, an imaging non-resonant galvanometer 10, an objective lens 16, a wavefront control device 21, a lens group and a plurality of lenses;

[0044] The system switches the system optical path to a multiphoton imaging optical path or an adaptive optics (AO) multiphoton imaging optical path by adjusting the position of the electrically controlled movable reflector, specifically including:

[0045] Multiphoton imaging optical path: the first electrically-controlled movable reflector 1 in the first position and the second electrically-controlled movable reflector 4 in the first position are not connected to the multiphoton imaging optical path, the output of the imaging laser directly acts on the imaging resonant galvanometer 6 after beam expansion through the lens group, the output of the imaging resonant galvanometer 6 acts on the imaging non-resonant galvanometer 10 after amplification by the lens, the output of the imaging non-resonant galvanometer 10 acts on the objective lens 16 after reflection by the dichroic mirror 11 through the first optical path containing the lens;

[0046] Adaptive optical multiphoton imaging optical path: the output of the imaging laser is reflected and directed by the first electrically-controlled movable reflector 1 in the second position, and then expanded by the lens group. The expanded light beam is regulated by the wavefront control device 21. The output of the wavefront control device 21 is reduced in spot size by the lens, and then reflected and adjusted by the second electrically-controlled movable reflector 4 in the second position to act on the imaging resonant galvanometer 6. The output of the imaging resonant galvanometer 6 is amplified by the lens and acts on the imaging non-resonant galvanometer 10. The output of the imaging non-resonant galvanometer 10 is reflected by the dichroic mirror 11 and acts on the objective lens 16 through the first optical path containing the lens; the output of the stimulation laser acts on the objective lens 16 through the laser stimulation optical path.

[0047] The optical path of the system is switched to a multi-photon light stimulation optical path or a multi-photon holographic light stimulation optical path by adjusting the position of the electrically controlled movable reflector, specifically:

[0048] Multi-photon light stimulation optical path: the output of the stimulation imaging laser is reflected and directed by the third electrically-controlled movable reflector 17 in the first position, and then expanded by the lens group. The expanded light beam is reflected and directed by the sixth electrically-controlled movable reflector 26 in the first position, and then acts on the stimulation non-resonant galvanometer 9. The output of the stimulation non-resonant galvanometer 9 is transmitted by the dichroic mirror 11 and then acts on the objective lens 16 through the first optical path containing the lens;

[0049] Multiphoton holographic light stimulation optical path: the third electrically-controlled movable reflector 17 in the second position, the fourth electrically-controlled movable reflector 19 in the second position, and the sixth electrically-controlled movable reflector 26 in the second position are not connected to the multiphoton holographic light stimulation optical path. The output of the stimulation imaging laser is expanded and acts on the wavefront control device 21. The output of the wavefront control device 21 is reflected and adjusted in direction by the fifth electrically-controlled movable reflector 23 in the second position, and then focused by a lens. After the zero-order light is blocked by the zero-order baffle 25 at the focus, the spot size is adjusted by the lens to be consistent with the size before entering the stimulation non-resonant galvanometer 9 in the multiphoton light stimulation optical path. The output of the stimulation non-resonant galvanometer 9 is transmitted through the dichroic mirror 11 and acts on the objective lens 16 through the first optical path containing the lens.

[0050] The multi-photon light stimulation optical path and the multi-photon holographic light stimulation optical path share the same optical path after acting on the non-resonant galvanometer 9 to stimulate the non-resonant galvanometer.

[0051] Specifically, in this embodiment, the imaging laser (Laser IMG) uses Spark Laser ALCOR 920, and the stimulation laser (Laser STI) uses Anyang FemtoYL TM -20 laser, the imaging resonant galvanometer 6 uses the Cambridgetechnology 8K resonant galvanometer, the imaging non-resonant galvanometer 10 uses the Cambridge technology6215 galvanometer, the stimulating non-resonant galvanometer 9 uses the Jinhaichuang SG1105 galvanometer, and the wavefront modulator 21 uses the meadowlark 1024×1024SpatialLight Modulator.

[0052] In this embodiment, when the electrically controlled movable reflector 1 is located at position ① and the electrically controlled movable reflector 4 is located at position ①, it is a common multi-photon imaging optical path. The imaging laser (Laser IMG) emits laser light (spot of about 1 mm), which is magnified 5 times by the lens group 2, and further enlarged 1.25 times by the lens 7 with a focal length of 100 mm and the lens 8 with a focal length of 125 mm after passing through the resonant galvanometer 6. Then, after the laser light passes through the non-resonant scanning galvanometer 10, it is reflected by the dichroic mirror 11, and further enlarged 2.5 times by the scanning lens 12 with a focal length of 80 mm and the sleeve lens 14 with a focal length of 200 mm. Finally, the light beam enters the objective lens 16. After multiple beam expansions, the light spot is magnified about 15 times before entering the objective lens, almost filling the objective lens entrance pupil, and high-resolution multi-photon imaging can be performed.

[0053] Adaptive optics (AO) is a technology used to compensate for aberrations in optical systems, which can significantly improve the imaging quality of microscopes. The AO system improves the focus and imaging clarity of the light beam by real-time monitoring and correction of optical aberrations, which is particularly important in deep tissue imaging.

[0054] In this embodiment, when the electrically controlled movable reflector 1 is located at position ②, the electrically controlled movable reflector 19 is located at position ①, the electrically controlled movable reflector 23 is located at position ①, and the electrically controlled movable reflector 4 is located at position ②, this is a multi-photon imaging optical path with adaptive optics. The imaging laser (Laser IMG) emits a laser (spot of about 1 mm), which is reflected by the electrically controlled movable reflector 1, and after the spot is magnified 3 times by the beam expander 3, it is reflected by the electrically controlled movable reflector 19, and after the spot is further magnified 5 times by the lens group 20, it enters the wavefront control device 21. At this time, the spot is magnified 15 times, almost filling the meadowlark 1024×1024 SLM 22 (Array Size 17.4×17.4mm), thereby maximizing the wavefront modulation effect. Then the light beam passes through the lens 22 with a focal length of 350 mm and the lens 5 with a focal length of 120 mm, and the spot is reduced to about 5 times the laser light outlet, ensuring that the spot size is close to the spot size before the ordinary multi-photon imaging optical path reaches the resonant galvanometer 6. After being reflected by the electrically controlled movable reflector 4, the optical path is merged with the ordinary multi-photon imaging optical path. After subsequent beam expansion, the spot size almost fills the objective lens entrance pupil, realizing high-resolution adaptive optical multi-photon imaging.

[0055] In this embodiment, when the electrically controlled movable reflector 17 is located at position ① and the electrically controlled movable reflector 26 is located at position ①, this is the ordinary multi-photon light stimulation optical path. The stimulation laser (Laser STI) emits a laser (spot size is about 3 mm), which is reflected by the electrically controlled movable reflector 17, magnified 1.5 times by the lens group 18, and reflected by the electrically controlled movable reflector 26, and then enters the light stimulation non-resonant galvanometer 9, which has a size of 5 mm, and the spot almost fills the entire galvanometer. The dichroic mirror 11 transmits the light stimulation laser, and after passing through the scanning lens 12 with a focal length of 80 mm and the sleeve lens 14 with a focal length of 200 mm, the spot is further magnified 2.5 times, and finally the light beam enters the objective lens. After multiple beam expansions, the spot is magnified by about 3.75 times before entering the objective lens, and high-precision multi-photon stimulation can be performed.

[0056] In this embodiment, when the electrically controlled movable reflector 17 is located at position ②, the electrically controlled movable reflector 19 is located at position ②, the electrically controlled movable reflector 23 is located at position ②, and the electrically controlled movable reflector 26 is located at position ②, it is a multi-photon holographic light stimulation optical path. The stimulation laser (Laser STI) emits a laser (spot size is about 3mm), which enters the wavefront control device 21 after being magnified 5 times by the lens group 20. At this time, the spot almost fills the meadowlark1024×1024SLM (Array Size 17.4×17.4mm), which maximizes the wavefront modulation effect. The laser modulated and reflected by the SLM is reflected by the electrically controlled movable reflector 23, passes through the lens 24 with a focal length of 350 mm, is blocked by the zero-order baffle 25 at the focusing point, and then passes through the lens 27 with a focal length of 150 mm. At this time, the spot size is about 2.15 times the size of the laser light outlet, occupying the stimulation non-resonant galvanometer 9, and then the optical path is merged with the ordinary multi-photon light stimulation. After multiple beam expansions, the spot is magnified by about 5.375 times before entering the objective lens, and high-precision multi-photon holographic light stimulation can be performed.

[0057] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can easily think of various equivalent modifications or replacements within the technical scope disclosed by the present invention, and these modifications or replacements should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be based on the protection scope of the claims.

Claims

1. A mode-switchable multiphoton imaging system, characterized in that: The invention comprises an imaging laser, a first electrically-controlled movable reflector (1), a second electrically-controlled movable reflector (4), a fourth electrically-controlled movable reflector (19), an imaging resonant galvanometer (6), an imaging non-resonant galvanometer (10), a first dichroic mirror (11), an objective lens (16), a wavefront control device (21), a lens and a plurality of lens groups; The system switches the system optical path to a multiphoton imaging optical path or an adaptive optical multiphoton imaging optical path by adjusting the position of the electrically controlled movable reflector, specifically comprising: Multiphoton imaging optical path: a first electrically controlled movable reflector (1) at a first position and a second electrically controlled movable reflector (4) at a first position are not connected to the multiphoton imaging optical path, an output of an imaging laser acts on an imaging resonant galvanometer (6), an output of the imaging resonant galvanometer (6) acts on an imaging non-resonant galvanometer (10), and an output of the imaging non-resonant galvanometer (10) acts on an objective lens (16) after being reflected by a first dichroic mirror (11); Adaptive optical multi-photon imaging optical path: the output of an imaging laser is reflected by a first electrically controlled movable reflector (1) at a second position to adjust the direction, then is adjusted by a fourth electrically controlled movable reflector (19) at a first position, and then is adjusted by a wavefront control device (21); the output of the wavefront control device (21) is reflected by a second electrically controlled movable reflector (4) at a second position to adjust the optical path and then acts on an imaging resonant galvanometer (6); the output of the imaging resonant galvanometer (6) acts on an imaging non-resonant galvanometer (10); the output of the imaging non-resonant galvanometer (10) is reflected by a first dichroic mirror (11) and then acts on an objective lens (16) through a first optical path containing a lens.

2. The system according to claim 1, characterized in that The multi-photon imaging optical path and the adaptive optical multi-photon imaging optical path share the same optical path after acting on the imaging resonance galvanometer (6).

3. A mode-switchable multiphoton stimulation system, characterized in that: The system comprises a stimulation laser, a third electrically-controlled movable reflector (17), a fourth electrically-controlled movable reflector (19), a fifth electrically-controlled movable reflector (23), a sixth electrically-controlled movable reflector (26), a stimulation non-resonant galvanometer (9), a first dichroic mirror (11), an objective lens (16), a wavefront control device (21), a lens, and a plurality of lens groups; The system switches the system optical path to a multi-photon light stimulation optical path or a multi-photon holographic light stimulation optical path by adjusting the position of the electrically controlled movable reflector, specifically: Multi-photon light stimulation optical path: the output of the stimulation imaging laser is reflected and adjusted in direction by a third electrically-controlled movable reflector (17) at a first position, and then acts on a stimulation non-resonant galvanometer (9) after being reflected and adjusted in direction by a sixth electrically-controlled movable reflector (26) at a first position; the output of the stimulation non-resonant galvanometer (9) is transmitted through a first dichroic mirror (11) and then acts on an objective lens (16) through a first optical path containing a lens; The multi-photon holographic light stimulation optical path: the third electrically controlled movable reflector (17) in the second position, the fourth electrically controlled movable reflector (19) in the second position, and the sixth electrically controlled movable reflector (26) in the second position are not connected to the multi-photon holographic light stimulation optical path; the output of the stimulation imaging laser acts on the wavefront control device (21) after beam expansion; the output of the wavefront control device (21) enters the stimulation non-resonance galvanometer (9) after being reflected and adjusted in direction by the fifth electrically controlled movable reflector (23) in the second position; the output of the stimulation non-resonance galvanometer (9) acts on the objective lens (16) through the first optical path containing a lens after being transmitted through the first dichroic mirror (11).

4. The system according to claim 3, characterized in that The multi-photon light stimulation optical path and the multi-photon holographic light stimulation optical path share the same optical path after acting on the non-resonant galvanometer (9) to stimulate the non-resonant galvanometer.

5. The system according to claim 3, characterized in that In the multi-photon holographic light stimulation light path, after the direction is adjusted by the fifth electrically-controlled movable reflector (23) in the second position, before entering the non-resonant galvanometer (9), the light path reflected by the fifth electrically-controlled movable reflector (23) is also included: the light path lens is focused, and the zero-order light is blocked at the focus by a zero-order baffle (25).

6. The system according to claim 1 or 3, characterized in that: A plurality of groups of lenses for adjusting the diameter of the light beam are arranged on the optical path.

7. The system according to claim 1 or 3, characterized in that: A scanning lens (12), a cylindrical lens (14) and a second dichroic mirror (15) are sequentially arranged on the first optical path containing a lens, and the output of the cylindrical lens (14) is reflected by the second dichroic mirror (15) and then acts on the objective lens (16).

8. The system according to claim 7, characterized in that The system also includes a PMT (28). The signal collected by the objective lens (16) is transmitted through the second dichroic mirror (15), and then separated into required wavelength signals by the third dichroic mirror (30) before acting on the PMT (28).

9. A mode-switchable multiphoton imaging and stimulation system, characterized in that: The system comprises an imaging subsystem and a stimulation subsystem; the imaging subsystem adopts the multiphoton imaging system described in claim 1, and the stimulation subsystem adopts the multiphoton stimulation system described in claim 3.

10. The system according to claim 9, characterized in that The multi-photon imaging optical path and the multi-photon stimulation system share the same optical path after passing through the first dichroic mirror (11).

Images