CN115826218B

CN115826218B - Multiphoton microscope host and multiphoton microscope system

Info

Publication number: CN115826218B
Application number: CN202211364119.8A
Authority: CN
Inventors: 吴润龙; 田景全; 云子艳; 胡炎辉; 王爱民; 程和平
Original assignee: Beijing Chaoweijing Biological Technology Co ltd; Peking University
Current assignee: Beijing Chaoweijing Biological Technology Co ltd; Peking University
Priority date: 2022-11-02
Filing date: 2022-11-02
Publication date: 2024-07-23
Anticipated expiration: 2042-11-02
Also published as: CN115826218A

Abstract

The application provides a multiphoton microscope host and a multiphoton microscope system, wherein the multiphoton microscope host comprises a mounting main body, and a wide-field searching module, a laser coupling module, a fluorescence collecting module and a scanning control module which are integrated on the mounting main body; the wide-field searching module is used for performing wide-field imaging on a living body so as to search a target area for installing a microscope probe on the living body; the laser coupling module is used for receiving the laser and adjusting the laser so as to couple the laser into the laser transmission optical fiber; the scanning control module is connected with the microscope probe through a control cable and used for controlling the microscope probe to perform laser scanning so as to generate a fluorescent signal; the fluorescence collection module is connected with the microscope probe through a fluorescence collection optical fiber and is used for collecting fluorescence signals output by the microscope probe. The technical scheme provided by the application has the advantages of small volume, easiness in carrying, convenience in use, easiness in installation and maintenance and the like.

Description

Multiphoton microscope host and multiphoton microscope system

Technical Field

The application relates to the technical field of optics, in particular to a multi-photon microscope host and a multi-photon microscope system.

Background

Direct recording of neuronal activity in free-moving animals is one of the most direct and effective methods to study the relationship between animal behavior and neural function, while multiphoton microscopy devices are the most important and widely used tool for observation of animal neurons using fluorescence imaging. The multiphoton microscope device may be a two-photon, three-photon or raman nonlinear laser scanning microscope device.

At present, the multiphoton microscope equipment has the problems of complex structure, large volume, occupied space, more wires, difficult transportation and transfer, complex field installation and later maintenance and the like.

Disclosure of Invention

In view of the above, the present application provides a multi-photon microscope host and a multi-photon microscope system, which are used for solving the problems of large occupied space, difficult transportation and transfer, complex installation and maintenance, etc. of the multi-photon microscope in the prior art.

The application provides a multiphoton microscope host machine, which is used for being connected with a microscope probe, and comprises a mounting main body, and a wide-field searching module, a laser coupling module, a fluorescence collecting module and a scanning control module which are integrated on the mounting main body; wherein,

The wide-field searching module is used for carrying out wide-field imaging on a living body so as to search a target area for installing a microscope probe on the living body;

the laser coupling module is used for receiving laser and adjusting the laser so as to couple the laser into a laser transmission optical fiber, wherein the laser transmission optical fiber is used for connecting the laser coupling module and the microscope probe;

the scanning control module is connected with the microscope probe through a control cable and used for controlling the microscope probe to perform laser scanning so as to generate a fluorescent signal;

The fluorescence collection module is connected with the microscope probe through a fluorescence collection optical fiber and used for collecting fluorescence signals output by the microscope probe.

Optionally, the multiphoton microscope further comprises a view finding adapter mounted on the mounting body;

The view finding adapter comprises a probe mounting assembly for detachably mounting the microscope probe and a switching mechanism arranged to enable switching of the probe mounting assembly to a first position and a second position;

When the probe mounting assembly is located at the first position, the microscope probe mounted on the probe mounting assembly avoids the optical path between the wide-field searching module and the living body, and when the probe mounting assembly is located at the second position, the microscope probe is aligned with the optical path of the wide-field searching module.

Optionally, the multi-photon microscope host further includes a moving module disposed on the mounting body, the moving module is configured to carry a living body and can drive the living body to move in multiple directions, and the wide-field searching module is configured to perform a field of view search on the living body located on the moving module.

Optionally, the multiphoton microscope host further includes a switchable light blocking door mounted on the mounting body;

When the light-shielding door is in a closed state, a closed space is formed between the mounting main body and the light-shielding door, and the mobile module is positioned in the closed space;

The wide field searching module is arranged to perform wide field searching on the living body on the moving module positioned in the closed space.

Optionally, the installation main body comprises a base and an installation frame fixed on the base, and a supporting plate is arranged at the upper part of the installation frame; wherein,

The mobile module is movably arranged on the base and is positioned on one side of the mounting frame, a control box positioned below the supporting plate is arranged on the other side of the mounting frame, and the scanning control module and the fluorescence collection module are arranged in the control box;

The laser coupling module is arranged above the supporting plate, and the wide-field searching module is arranged above the moving module.

Optionally, the wide-field searching module is installed on the laser coupling module, an optical path through hole penetrating up and down is arranged on the laser coupling module, and an optical path of the wide-field searching module is arranged to penetrate through the optical path through hole downwards to reach the mobile module.

Optionally, the laser coupling module includes a coupler housing, a dispersion compensating element, an acousto-optic modulator, and a beam stabilizing device, where the dispersion compensating element, the acousto-optic modulator, and the beam stabilizing device are all disposed in the coupler housing and are sequentially disposed along a transmission direction of laser; wherein,

The dispersion compensation element is used for compensating negative dispersion caused by the laser transmission process of the transmission fiber;

the acousto-optic modulator is used for adjusting the intensity of laser;

the beam stabilizing device is used for adjusting the laser transmission direction so as to correct the deviation between the actual position of the laser beam at the laser output end of the laser coupling module and the ideal position.

Optionally, the laser coupling module further comprises a driver for driving the acousto-optic modulator and a cooling mechanism for cooling the driver, both of which are located on the upper surface of the coupler housing.

Optionally, the wide-field searching module is installed on the upper surface of the coupler shell, the laser coupling module is provided with a light path through hole penetrating up and down, and a light path of the wide-field searching module is arranged to penetrate through the light path through hole downwards;

the multiphoton microscope host also includes a cover for covering the wide field search module, the driver, and the cooling mechanism.

Optionally, the multiphoton microscope host further includes a control box mounted on the mounting body, and the fluorescence collection module and the scanning control module are both located in the control box.

Optionally, a first interface for connecting the fluorescence collection optical fiber and a second interface for connecting the control cable are arranged on the control box, and the first interface and the second interface are both positioned above the same side of the control box;

the laser coupling module is located above the control box, and the output end of the laser coupling module, which is used for connecting the laser transmission optical fiber, is located on the same side as the first interface and the second interface of the control box.

Optionally, the multiphoton microscope further comprises a receiving device mounted on a side of the control box having the first interface and the second interface;

The storage device is used for storing the microscope probe and a cable which is connected with the microscope probe and comprises the laser transmission optical fiber, the fluorescence collection optical fiber and the control cable.

According to another aspect of the present application, there is also provided a multiphoton microscope system including: a laser, a microscope probe and a multiphoton microscope mainframe as described above;

The laser is arranged to transmit laser to the laser coupling module, the output end of the laser coupling module is connected with the microscope probe through a laser transmission optical fiber, the fluorescence collecting module is connected with the microscope probe through a fluorescence collecting optical fiber, and the scanning control module is connected with the microscope probe through a control cable.

In the technical scheme provided by the application, the multi-photon microscope host integrates all the functional modules into a whole structure, so that the occupied space can be greatly reduced, the multi-photon microscope host is suitable for various laboratories, and the whole multi-photon microscope host is arranged to enable the wire outlet to be regular, neat and attractive. In addition, because the multiphoton microscope host computer is small, portable, easily transport and change the place, can adjust the position and the direction of this multiphoton microscope fast to some experimental demands simultaneously, conveniently match more applications. In addition, the multiphoton microscope host is convenient for on-site rapid installation and maintenance.

Additional features and advantages of the application will be set forth in the detailed description which follows.

Drawings

The accompanying drawings, which are included to provide a further understanding of the application, illustrate and explain the application and are not to be construed as limiting the application. In the drawings:

FIG. 1 is a schematic diagram of the external structure of a multi-photon microscope host according to one embodiment of the application;

FIG. 2 is a schematic view of the structure of the light-shielding door of the host computer of the multi-photon microscope shown in FIG. 1 in an open state;

FIG. 3 is a schematic diagram of a multi-photon microscope in an exploded state according to one embodiment of the application;

FIG. 4 is a schematic diagram of a multi-photon microscope mainframe in an exploded state from another perspective;

FIG. 5 is a schematic diagram of the mating installation of the mobile module, the living body mounting device, and the view finding adapter according to one embodiment of the present application;

fig. 6 is a schematic structural view of a living body mounting device according to an embodiment of the present application;

FIG. 7 is a schematic diagram of a laser coupling module according to one embodiment of the application;

FIG. 8 is a schematic diagram of the internal structure of the laser coupling module shown in FIG. 7;

FIG. 9 is a schematic diagram of a wide field search module mounted on a laser coupling module according to one embodiment of the present application;

FIG. 10 is a schematic diagram of a control box in an embodiment in accordance with the application;

FIG. 11 is a front view of the control box shown in FIG. 10;

Fig. 12 is a schematic structural view of a storage device according to an embodiment of the present application;

Fig. 13 is a schematic view showing a structure in which the storage device is in an exploded state according to an embodiment of the present application;

FIG. 14 is a schematic diagram of a multiphoton microscope system according to an embodiment of the application;

fig. 15 is a schematic structural diagram of a multiphoton microscope system according to another embodiment of the present application.

Reference numerals illustrate:

100-multiphoton microscope hosts; 1-mounting a main body; 11-a base; 12-mounting frame; 13-a support plate; 14-a handle; 15-a display screen; 2-a light-shielding door; a 3-laser coupling module; 31-input terminal; 32-an output; 311-a power detector; 33-a coupler housing; 331-optical path through holes; a 34-dispersion compensating element; 35-a mirror; 36-an acousto-optic modulator; 361-a driver; 362-heat dissipating fins; 363-fan; 37-a first deflection mirror; 38-a second deflection mirror; 39-position detector; 4-a visual field searching module; 41-a fluorescent light source; 42-a camera; 43-objective lens; 5-a control box; 51-first interface; 52-a second interface; 53-a fluorescence collection module; 54-a main control circuit board; 6-a storage device; 61-a storage box; 611-a projection; 612-through holes; 613-an annular groove; 614-through slots; 62-ring indicator lights; 63-a wire baffle disc; 631-a light-transmitting cover; 632-wire blocking ring; 633-winding reel; 64-clamp wire loops; 643-a slot; 65-probe holder; 66-protective cover; 661-an annular cover; 662-a viewing window; 67-rotating shaft; 68-hinges; 7-a mobile module; 8-living body mounting means; 81-mounting seats; 811-a baffle; 812-

A fixing frame; 82-a clamping assembly; 821-first adjusting bolt; 83-a light shield; 84-rotating the component; 85-moving rack; 86-a second adjusting bolt; 87-a third adjusting bolt; 88-a treadmill; 89-receiving the tray; 80-probe mount; 9-a view search adapter; 91-a probe mounting assembly; 10-cover cap; 200-a laser; 300-a laser adapter; 301-transmission optical fiber; 400-microscope probe; 401-a laser transmission fiber; 402-fluorescence collection fiber; 403-control cable.

Detailed Description

The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application. Embodiments of the present application and features of the embodiments may be combined with each other without conflict.

In the description of the present application, it should be understood that the terms "center," "longitudinal," "transverse," "upper," "lower," "left," "right," "vertical," "horizontal," "top," "bottom," "axial," "radial," "circumferential," and the like indicate orientations or positional relationships based on the orientation or positional relationships shown in the drawings, merely to facilitate describing the present application and simplify the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and therefore should not be construed as limiting the present application. In addition, "inner and outer" refer to inner and outer with respect to the outline of each component itself.

Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature.

The application provides a multiphoton microscope host machine, which is used for being connected with a microscope probe, wherein the microscope probe is used for being detachably arranged on a living body so as to observe neuron activities of the living body.

As shown in fig. 1 to 4, a multiphoton microscope host 100 provided by an embodiment of the present application includes a mounting body 1, and a wide field searching module 4, a laser coupling module 3, a fluorescence collecting module, and a scan control module integrated on the mounting body 1; wherein,

The wide field searching module 4 is configured to perform wide field imaging on a living body to search a target area on the living body for mounting a microscope probe;

the laser coupling module 3 is configured to receive the laser light and adjust the laser light to couple the laser light into a laser transmission fiber, wherein the laser transmission fiber is configured to connect the laser coupling module 3 and the microscope probe;

The scanning control module is connected with the microscope probe through a control cable and used for controlling the microscope probe to carry out laser scanning on a living body so as to generate a fluorescent signal;

The fluorescence collection module is connected with the microscope probe through a fluorescence collection optical fiber and is used for collecting fluorescence signals output by the microscope probe.

In application, the multiphoton microscope host 100 is connected to the microscope probe 400, and referring to fig. 14 and 15, the laser coupling module 3 of the multiphoton microscope host 100 is connected to the microscope probe 400 through the laser transmission fiber 401, the scanning control module is connected to the microscope probe 400 through the control cable 403, the fluorescence collection module is connected to the microscope probe 400 through the fluorescence collection fiber 402, and the microscope probe 400 is used for being worn on a living body. When the laser coupling module 3 receives laser and transmits the laser to the microscope probe 400 through the laser transmission optical fiber 401, the scanning control module controls the microscope probe 400 to perform laser scanning on a living body through the control cable 403 to generate a fluorescent signal, the fluorescent collecting module collects the fluorescent signal output by the microscope probe 400 through the fluorescent collecting optical fiber 402, the output fluorescent signal can be converted into an electric signal and imaged on a computer, and then the neuron activity condition of the living body is observed through imaging.

The microscope probe 400 may include a microelectromechanical (MEMS, micro-Electro-MECHANICAL SYSTEM) scanning galvanometer, among others, as well as various lenses. Thus, the scanning control module may comprise a MEMS control module that controls the microelectromechanical scanning galvanometer.

The multiphoton microscope host provided by the application integrates all the functional modules to form a whole structure, can greatly reduce occupied space, is suitable for various laboratories, is arranged as a whole machine, and can lead the outgoing lines to be regular, neat and beautiful. In addition, the multi-photon microscope host is small in size, portable, easy to carry and replace, and capable of quickly adjusting the position and the direction of the multi-photon microscope according to some test requirements, and more applications can be conveniently matched. In addition, the multiphoton microscope host is convenient for on-site rapid installation and maintenance. The multiphoton microscope device can be a two-photon, three-photon or Raman nonlinear laser scanning microscope.

In one embodiment, the multiphoton microscope host 100 further includes a moving module 7 disposed on the mounting body 1, the moving module 7 is configured to carry a living body and can drive the living body to move in multiple directions, and the wide-field searching module 4 is configured to perform a view search on the living body located on the moving module 7.

Specifically, the living body may be directly mounted on the mobile module 7, and the living body may be specifically restricted by providing a clamping or limiting structure or the like on the mobile module 7. It is also possible to first mount the living body on the living body mounting device 8 and then fix the living body mounting device 8 on the moving module 7, and the moving module 7 can move with the living body mounting device 8 to adjust the position of the living body, whereby the wide-field searching module 4 can image different areas of the living body to search for the target position of interest. The movement module 7 may be a multi-axis movement platform, and may be movable in a plurality of directions, i.e., up, down, left, right, front, and rear.

The wide-field searching module 4 is a device capable of imaging a wide-field area of a living body, and can use single photon fluorescence imaging. The imaging of the wide-field searching module 4 can be transmitted to a computer for display, or an eyepiece can be arranged on the wide-field searching module 4, and imaging can be directly observed from the eyepiece.

In the embodiment shown in fig. 5, when the wide field imaging of the living body is performed using the wide field searching module 4, the living body is mounted on the living body mounting device 8, and then the living body mounting device 8 is fixed on the moving module 7. The living body may be a mouse or other animal.

Fig. 6 provides a living body mounting device 8 suitable for mounting a mouse (not excluding for mounting other suitable animals), comprising: mount 81, treadmill 88 and fixture are all installed on mount 81. The clamping mechanism may include two oppositely disposed clamping assemblies 82 for clamping both sides of a probe mount 80 disposed on a mouse (the probe mount 80 is typically intended to be mounted on the head of a mouse, and fig. 6 only shows the probe mount 80 and not the mouse). The clamping mechanism is configured to enable a mouse to run on the treadmill 88 while the probe mount 80 is clamped and secured. Wherein, the mounting seat 81 is provided with baffle plates 811 positioned at two sides of the running machine 88 for limiting the mice on the running machine 88, and the mounting seat 81 is also provided with a fixing frame 812 fixed on the mobile module 7 through bolts.

When the probe mounting piece 80 arranged on the mouse is clamped by the clamping component 82, the mouse runs on the running machine 88, so that the attention of the mouse can be dispersed, the stress response of the awake mouse in the mounting process is reduced, and the quick experiment is facilitated.

Optionally, each clamping assembly 82 comprises two clamping portions and a first adjustment bolt 821, the first adjustment bolt 82 being arranged to enable the two clamping portions to be brought closer to each other to clamp the probe mount 80, or to be brought away from each other to release the probe mount 80, by rotation. Therefore, when the probe mount 80 is clamped or disassembled, only the first adjusting bolts 821 of the two clamping assemblies 82 need to be rotated, and the assembly and the disassembly are convenient.

Alternatively, the two clamping assemblies 82 are provided to be movable in the front-rear direction of the treadmill 88 with respect to the mount 81 and to be movable up and down with respect to the mount 81. In this way, the clamping assembly 82 can be adjusted to position to clamp the probe mount 80 according to the type or size of living subject.

Specifically, the living body fixing device 8 further includes a moving frame 85 provided corresponding to each of the clamping assemblies 82, the moving frame 85 being provided so as to be movable in the front-rear direction of the treadmill 88 with respect to the mount 81, the clamping assemblies 82 being liftably provided on the moving frame 85. As shown in fig. 6, a second adjusting bolt 86 for adjusting the lifting of the clamping assembly 82 is provided on the moving frame 85, the clamping assembly 82 can be lifted up by rotating the second adjusting bolt 86, the second adjusting bolt 86 is reversely rotated, and the clamping assembly 82 can be lowered under the action of gravity. The mount 81 is provided with a third adjusting bolt 87 for adjusting the forward and backward movement of the moving frame 85. The third adjusting screw 87 is in threaded connection with the movable frame 85, and the movable frame 85 can be made to move back and forth relative to the mounting seat 81 by rotating the third adjusting screw 87.

Optionally, the living body fixing device 8 further includes a light shielding mechanism for shielding the living eye. The light shielding mechanism includes a light shielding cover 83 and a rotating member 84 that drives the light shielding cover 83 to rotate. In wide field searching imaging of mice, the rotating member 84 may be operated such that the eye shield 83 shields the eyes of the mice to protect the eyes from light.

The living body fixing device 8 may further include a water fountain (not shown) disposed on the mounting base 81, and the water fountain is configured to provide water for the mice on the running machine, so as to facilitate the distraction of the mice.

The living body fixing device 8 may further include a receiving tray 89 provided below the mount 81 for receiving excreta of the mice.

In one embodiment, as shown in fig. 3 and 5, the multiphoton microscope host further includes a view finding adapter 9 mounted on the mounting body 1; the view finding adapter 9 includes a probe mount assembly 91 for detachably mounting the microscope probe 400, and a switching mechanism configured to be able to switch the probe mount assembly 91 to a first position and a second position.

When the probe mounting assembly 91 is located at the first position, the microscope probe 400 mounted on the probe mounting assembly 91 avoids the optical path between the wide-field searching module 4 and the living body, and when the probe mounting assembly 91 is located at the second position, the microscope probe 400 is aligned with the optical path of the wide-field searching module 4.

Wherein, the switching mechanism may be configured to switch the probe mounting assembly 91 between two positions by manually pushing and pulling, specifically, a holding portion (not shown in the figure) that is convenient to hold may be disposed on the switching mechanism, and when the probe mounting assembly 91 is pushed by the holding portion, the probe mounting assembly 91 may be moved to the first position, and when the probe mounting assembly 91 is pulled in the opposite direction, the probe mounting assembly 91 may be moved to the second position.

Further, the field searching adaptor 9 may be provided with an objective lens 43, and when the field searching adaptor 9 is mounted on the mounting body 1, the objective lens 43 is aligned with the optical path of the wide field searching module 4, and when the probe mounting assembly 91 is in the second position, the microscope probe 400 mounted thereon is aligned with the optical path of the wide field searching module 4. When the wide field imaging is performed by using the wide field searching module 4, the probe mounting assembly 91 is switched to the first position by using the switching mechanism, after the target area is found on the living body by the wide field searching module 4, the probe mounting assembly 91 is switched to the second position, and then the microscope probe 400 on the probe mounting assembly 91 is removed and fixed at a position (which can be fixed by means of gluing) of the probe mounting member 80 of the living body corresponding to the found target area.

After the microscope probe 400 is mounted on the probe mount 80, a living body (e.g., a mouse) can be detached from the living body mounting device 8, and the mouse is released to be free to move, whereby neuron observation can be performed on the free-moving mouse through the microscope probe 400.

In one embodiment, the multiphoton microscope host 100 further includes a light shielding door 2 capable of being opened and closed mounted on the mounting body 1. When the light-shielding door 2 is in a closed state, a closed space is formed between the mounting main body 1 and the light-shielding door 2, the moving module 7 is positioned in the closed space, and the wide-field searching module 4 is arranged to perform wide-field searching on a living body positioned on the moving module 7 in the closed space.

The arrangement of the light shielding door 2 can ensure that the living body is positioned in a darkroom when the wide-field searching module 4 images the living body, can achieve higher imaging signal to noise ratio, and the arrangement of the light shielding door 2 does not need to build a special light shielding environment (such as a large hood, or a lighting lamp for closing a laboratory, etc.).

Alternatively, as shown in fig. 1 and 2, the two light-shielding doors 2 may be provided in a split manner, that is, the two light-shielding doors 2 are respectively rotatably mounted on the mounting body 1 so as to be able to be closed when rotated toward each other and to be rotated away from each other. Fig. 1 shows a state where two light-shielding doors 2 are closed, and fig. 2 shows a state where two light-shielding doors 2 are open.

It will be appreciated that the light-shielding door 2 may be provided with only one, and may be provided to be opened and closed by lifting or sliding.

In one embodiment, the specific arrangement of the modules of the multi-photon microscope host 100 can be shown with reference to fig. 3 and 4, the mounting body 1 includes a base 11 and a mounting frame 12 fixed on the base 11, and a support plate 13 is disposed at an upper position of the mounting frame 12;

The mobile module 7 is movably arranged on the base 1 and is positioned on one side of the mounting frame 12, the other side of the mounting frame 12 is fixedly provided with a control box 5 positioned below the supporting plate 13, and the scanning control module and the fluorescence collection module are arranged in the control box 5; the laser coupling module 3 is mounted on a support plate 13, and the wide field searching module 4 is mounted above the moving module 7. The light shielding door 2 is mounted on a side of the mounting frame 12 facing the moving module 7, and is used for forming a closed space on a side of the mounting frame 12 having the moving module 7, and an optical path of the wide-field searching module 4 positioned above can enter the closed space so as to be capable of imaging a living body on the moving module 7.

Optionally, the wide-field searching module 4 is mounted on the laser coupling module 3, and the laser coupling module 3 is provided with an optical path through hole 331 penetrating up and down, and the optical path of the wide-field searching module 4 is set to penetrate down through the optical path through hole 331 to reach the moving module 7. The arrangement can make the whole structure more compact and the volume more compact.

Optionally, the base 11 may be provided with a handle 14, and the handle 14 may facilitate the carrying of the multiphoton microscope host 100.

Optionally, a display screen 15 may be further disposed on the base 11, and the display screen 15 may display: for example, parameters of laser, transmission state, temperature and humidity of a multiphoton microscope and the like, so that the working state of the equipment can be conveniently known.

In one embodiment, as shown in fig. 7 and 8, the laser coupling module 3 includes a coupler housing 33, a dispersion compensating element 34, an acousto-optic modulator 36, and a beam stabilizing device, where the dispersion compensating element 34, the acousto-optic modulator 36, and the beam stabilizing device are all disposed in the coupler housing 33 and are disposed in sequence along the transmission direction of the laser light; wherein,

The dispersion compensation element 34 is used to compensate for negative dispersion caused by the laser transmission fiber 401 during transmission of the laser light;

The acousto-optic modulator 36 is used for adjusting the intensity of the laser;

The beam stabilizing means are used to adjust the laser transmission direction to correct the deviation of the actual position of the laser beam at the output 32 of the laser coupling module 3 from the ideal position.

The beam stabilizing means may particularly comprise a position.

Wherein a position detector 39 is arranged close to the output 32 of the laser coupling module 3 for detecting the position information of the laser at the output 32. The position detector 39 may be a 4D position detector, and the 4D position detector can strictly detect the position drift and the angle drift of the distinguishing beam, and can accurately detect the real-time position of the beam. The reflector adjusting mechanism is configured to drive the deflection reflector to deflect according to the position information detected by the position detector 39 so as to adjust the laser transmission direction, so that the laser is stably output from the output end to the laser transmission fiber, and the coupling efficiency is improved.

Specifically, the ideal position of the laser beam at the output end 32, which is a position where the ideal coupling efficiency can be achieved when the laser is coupled into the connected laser transmission fiber 401 at the output end 32, is first determined. When the beam is deflected, for example, the beam is deflected due to deflection of an optical device caused by vibration, temperature change, or the like, or the beam is deflected due to artificial touch, or the like, the position detector 39 detects the position information of the laser at the output end 32 in real time and sends the position information to the control unit, and the control unit continuously determines the deviation of the position of the laser beam from the ideal position according to the position information, and controls the mirror adjusting mechanism to adjust the deflection mirror, thereby continuously adjusting the reflection direction of the laser so that the laser is stably transmitted to the laser transmission optical fiber 401 within a certain range around the ideal position. The control unit may be disposed in the laser coupler housing 33 or in the control box 5.

Optionally, the laser coupling module 3 may also be provided with at least one mirror 35 for changing the direction of laser transmission. By changing the laser transmission direction, the light path can be bent, so that the arrangement of components on the light path is facilitated, and the size of the whole laser coupling module is reduced.

As shown in fig. 8, the laser enters from the input end 31 of the laser coupling module 3, is subjected to dispersion compensation by the dispersion compensation element 34, and is transmitted to the reflecting mirror 35, is deflected by about 90 degrees by the reflecting mirror 35, and is transmitted to the acousto-optic modulator 36, the intensity of the laser is adjusted by the acousto-optic modulator 36, and is transmitted to the first deflecting mirror 37, is deflected by about 90 degrees by the first deflecting mirror 37, is reflected to the second deflecting mirror 38, is reflected to the output end 32 by the second deflecting mirror 38, and is coupled into the laser transmission fiber connected to the output end 32. The first deflecting and reflecting laser 37 and the second deflecting and reflecting mirror 38 are respectively provided with a corresponding mirror adjustment mechanism, and the positions of the first deflecting and reflecting laser 37 and the second deflecting and reflecting mirror 38 are adjusted in real time according to the position information detected by the position detector 39, so that the laser can be stably output to the laser transmission optical fiber 401.

A power detector 311 may also be provided in the laser coupling module 3 for detecting the laser transmission power.

In addition, as shown in fig. 9, the laser coupling module further includes a driver 361 for driving the acousto-optic modulator 36 and a cooling mechanism for cooling the driver 361, both of which are located on the upper surface of the coupler housing 33. The cooling mechanism may include a heat radiation fin 362 that radiates heat from the driver 361, and a fan 363 that radiates heat from the heat radiation fin 362.

Because the driver 361 drives the rf power to be large, the risk of high-power rf signal interference is increased in the laser coupler housing 33, and because the driver 361 generates a large amount of heat, the deformation of a precise optical system plate is easily caused, the temperature in the cavity is increased, and the performance of the device is affected, so that the driver 361 is arranged outside the coupler housing 33, and the heat dissipation fins 362 and the fans 363 are added to dissipate the heat.

Alternatively, referring to fig. 9, a wide field searching module 4 is installed on the upper surface of the coupler housing 33, and a light path through hole 331 penetrating up and down is provided on the laser coupling module 3, and a light path of the wide field searching module 4 is disposed to penetrate down through the light path through hole 331. The wide-field searching module 4 may include a fluorescent light source 41 and a camera 42, where the light path of the camera 42 passes through the light path through hole 331 downwards to reach the living body on the mobile module 7, so as to realize wide-field imaging on the living body.

The multiphoton microscope host may further include a cover 10 for covering the wide field searching module 4, the driver 361, and the cooling mechanism, and the cover 10 not only protects but also is attractive.

In one embodiment, as shown in fig. 10 and 11, the multiphoton microscope host further includes a control box 5 mounted on the mounting body 1, and the fluorescence collection module and the scanning control module are both located in the control box 5.

The fluorescence collection module 53 may include a photomultiplier tube (PMT), to which the signal collected by the fluorescence collection fiber 402 is transmitted.

Optionally, the fluorescence collection module 53 includes a beam splitter and at least two beam splitting and collecting modules, each of which may include a photomultiplier tube. The fluorescence signal collected by the fluorescence collection optical fiber 402 from the microscope probe 400 is split into at least two fluorescence signals by the spectroscope, and then is collected by at least two spectroscope collection modules respectively.

The control box 5 may further be provided with a signal processing module, where the signal processing module is configured to process a signal output by the fluorescent collection module 53 and transmit the processed signal to a computer for display. For example, the fluorescent signal collected by the fluorescent collection module 53 is converted into an electric signal, and the signal is amplified, then collected and recombined by high-speed AD collection, and then transmitted to a computer for display.

The fluorescence acquisition module and the signal processing module are both arranged in the control box 5, so that the transmission distance from the signal acquired by the fluorescence acquisition module to the signal processing module is shortened, the possibility of being interfered is reduced, and the reliability of signal transmission is improved.

Optionally, the control box 5 is provided with a first interface 51 for connecting the control cable 403 and a second interface 52 for connecting the fluorescence collection optical fiber 402, and the first interface 51 and the second interface 52 are located above the same side of the control box 5.

The laser coupling module 3 is located above the control box 5, and the output end of the laser coupling module 3 for connecting the laser transmission optical fiber 401 is located on the same side as the first interface 51 and the second interface 52 of the control box 5.

The laser coupling module 3 is arranged above the control box 5, the first interface 51 and the second interface 52 of the control box 5 are positioned above the box body, and the output end of the laser coupling module 3 and the first interface 51 and the second interface 52 of the control box 5 are positioned on the same side, so that the laser transmission optical fiber 401, the fluorescence collection optical fiber 402 and the control cable 403 are close to each other, the wiring is regular and attractive, the cables can be converged into a bus cable, for example, the cables can be wrapped into the bus cable through a cable cover after being converged, and the cables are conveniently stored through the storage device 6 (the storage of the storage device and the microscope probe will be described in detail).

In addition, the outgoing ends of the laser transmission optical fiber 401, the fluorescence collection optical fiber 402 and the control cable 403 are arranged above the control box 5, so that more behavioural devices can be conveniently adapted, and the cable length can be reduced. For example, when a mouse with the microscope probe 400 is placed in the living body behavior box to freely move, the arrangement of the cable can facilitate the microscope probe 400 to extend downwards into the living body behavior box.

Further, as shown in fig. 10 and 11, a main control circuit board 54 and a fluorescence collecting module 53 are disposed in the control box 5, and the fluorescence collecting module 53 is located above the main control circuit board 54, where the main control circuit board 54 includes a scan control module, and may further include a control driving circuit for controlling the laser coupling module 3 (e.g. controlling an acousto-optic modulator and a beam stabilizer of the laser coupling module 3), the view searching module 4, and an indicator light, an optical sensor, a temperature and humidity sensor, etc.

In one embodiment, the multiphoton microscope further comprises a receiving device 6, and the receiving device 6 is mounted on a side of the control box 5 having the first interface 51 and the second interface 52, and since the laser transmission fiber 401, the fluorescence collection fiber 402 and the control cable 403 are all located on the side, the receiving device 6 is mounted on the side to conveniently receive the microscope probe 400 and the cable connected to the microscope probe 400, including the laser transmission fiber 401, the fluorescence collection fiber 402 and the control cable 403. For ease of storage, the cable may be collectively wrapped, for example, with a wire wrap.

As shown in fig. 12 and 13, the storage device 6 includes:

A storage body provided with a winding drum 633, and an annular space for accommodating the cable is formed around the winding drum 633;

A probe holder 65, the probe holder 65 is fixed to the housing body, and the microscope probe 400 is mountable to the probe holder 65 after the cable is wound around the winding drum 633.

Fig. 12 shows a state in which the cable is wound on the winding drum 633 and the microscope probe 400 is mounted on the probe holder 65. The cable and the probe are not easily touched and pressed to cause damage by being stored through the storage device 6, the situation that the cable is randomly placed to cause winding and knotting and the situation that the cable is randomly bent to cause damage can be avoided, and the cable and the probe are received through the storage device 6 more neatly and attractive, so that the visual effect is promoted.

Specifically, the storage body may include: the probe holder 65 is fixed to the outside of the wire tray 63 facing away from the storage box 61. The winding drum 633 is provided on one of the storage box 61 and the wire blocking drum 62, and the storage box 61 and the wire blocking drum 63 are provided so as to be able to stop the cable wound on the winding drum 633 from both ends of the winding drum 633.

In the embodiment shown in fig. 13, the wire blocking disc 63 includes a winding drum 633 and a wire blocking ring 632 radially protruding from the winding drum 633, and when the winding drum 633 is fixed to the storage box 61, an annular space is formed between the wire blocking ring 632 and the storage box 61 around the winding drum 633. Of course, the winding drum 633 may be directly formed on the receiving box 61, and the wire blocking disc 63 is fixed at an end of the winding drum 633 facing away from the receiving box 61 and has a wire blocking ring 632 radially protruding from the winding drum 633.

The probe support 65 may have various structures, so long as the microscope probe 400 can be installed, for example, the probe support 65 may be configured as a clamping structure capable of clamping the probe, and a jack may be provided on the probe support 65, so that the microscope probe 400 is inserted into the jack.

Optionally, a plurality of limiting notches are circumferentially and alternately arranged on the outer circumference of the wire blocking disc 63 (i.e. on the wire blocking ring 632), and the cable extends from the winding drum 633 to the outer side of the wire blocking disc 63 so that when the probe is mounted on the probe support 65, the cable is limited by one of the limiting notches;

optionally, a wire loop 64 may be disposed on the outer side of the wire blocking disc 63 facing away from the winding drum 633, and a plurality of slots 643 disposed around the probe support 65 may be disposed on the wire loop 64, where a plurality of bumps disposed at intervals along the circumferential direction may be disposed on the wire loop 64, and the slots 643 are formed between adjacent bumps, respectively. When the cable extends to the outer side of the wire blocking disc 63 to enable the probe to be mounted on the probe support 65, the cable can be clamped in one of the clamping grooves 643, so that the cable can be prevented from loosening from the winding drum 633, and the situation that the probe is driven to be separated from the probe support due to swinging or loosening of the cable can be avoided. The wire loop 64 may be made of a flexible material, such as a rubber material, and the flexible material has elasticity that allows the cable to be easily clamped into and removed from the slot.

In one embodiment, the housing 61 is formed with a through hole 612 on a side thereof for abutting against the control box 5, the through hole 612 communicates with the annular space accommodating the cable, and an end of the cable having the microscope probe 400 enters the housing 61 from the through hole 612 and can be wound on the winding drum 633 while being protruded to the annular space.

Specifically, the housing 61 is provided with a projection 611 projecting toward the winding drum 633 around the through hole 612, and an annular groove 613 is formed on the radially outer side of the projection 611 away from the through hole 612; the wire tray 63 is provided with a winding drum 633, the winding drum 633 is fixed to the projection 611, and the annular groove 613 forms an annular space for winding the wire between the receiving box 61 and the wire tray 63. The protruding portion 611 may be provided with an annular structure, or may be provided with an arc structure with a notch, for installing the winding drum 633, an external thread may be provided on the outer surface of the protruding portion 611, an internal thread may be provided on the inner surface of the winding drum 633, and the winding drum 633 may be connected to the protruding portion 611 of the receiving box 61 by means of a threaded connection.

The projection 611 is provided with a through groove 614 penetrating the projection 611 radially, and the end of the cable having the microscope probe 400 can be extended from the through groove 614 to the annular space after entering the storage case 61 from the through hole 612. The through groove 614 may be a notch formed in the projection 611 as shown in fig. 13, or may be a through hole provided in a wall of the projection 611. When the winding drum 633 is fixed to the projection 611, the through groove 614 is located at substantially one end of the winding drum 633, and the cable can be wound around the winding drum 633 by extending from the through groove 614 to the annular space.

In one embodiment, the housing device further includes an annular indicator light 62, the annular indicator light 62 is mounted on the housing body, and the annular indicator light 62 is disposed around the center of the winding drum 633. The annular indicator lamp 62 may be configured to illuminate the interior of the storage device 6, or may be configured to indicate an operation state of the interior of the multi-photon microscope host, for example, the annular indicator lamp 62 may be configured to indicate that the apparatus is in an operation state, a failure in a non-operation state, or an abnormality in the apparatus, etc. using different colors. If the laser is detected to enter the laser coupling module 3 or the laser is detected to emit laser light, the controller can control the annular indicator lamp 62 to display green, and the green indicator device is in an operating state; when detecting an abnormality of the device, such as an abnormality of laser power, or other abnormal state, the annular indicator lamp 62 can be controlled to display red color for warning; while the device is in a non-operational state, the annular indicator light 62 may display a yellow color for interior illumination.

The wire blocking disc 63 includes a light-transmitting cover 631, and the annular indicator lamp 62 is disposed between the housing 61 and the wire blocking disc 63, and the lamp light of the annular indicator lamp 62 can be seen through the light-transmitting cover 631 corresponding to the light-transmitting cover 631. The wire ring 632 of the wire blocking disk 63 is disposed on the outer circumference of the light-transmitting cover 631.

In addition, the storage device 6 may further include a protective cover 66 covering the outside of the storage body, and the protective cover 66 is provided rotatably mounted on the storage body through a rotation shaft 67 and a hinge 68, and is rotatable to an open state and a closed state. To keep the protective cover 66 in the closed state, a magnet may be provided between the protective cover 66 and the storage main body.

The protective cover 6 may include an annular cover 661 and a transparent viewing window 62 provided at a central portion of the annular cover 661 to facilitate viewing of the condition inside the storage device, and to facilitate viewing of the status of the apparatus indicated by the annular indicator lamp 62.

In use, the probe is mounted on a living body for use by rotating the protective cap 66 open, removing the microscope probe 400 from the probe holder 65, and unwinding the cable from the winding drum 633.

According to another aspect of the present application, there is also provided a multiphoton microscope system, as shown in fig. 14 and 15, including: a laser 200, a microscope probe 400, and a multiphoton microscope host 100 as described above;

The laser 200 is configured to transmit laser light to the laser coupling module 3, the output end of the laser coupling module 3 is connected to the microscope probe 400 through a laser transmission optical fiber 401, the fluorescence collection module is connected to the microscope probe 400 through a fluorescence collection optical fiber 402, and the scanning control module is connected to the microscope probe 400 through a control cable 403.

In one embodiment, as shown in fig. 14, the multiphoton microscope system further includes a laser adapter 300, and the laser 200 first emits laser light to the laser adapter 300, and after the laser light is adjusted and adapted by the laser adapter 300, the laser light is transmitted to the multiphoton microscope host 100 through a transmission optical fiber 301.

By providing the laser adapter 300 between the laser 200 and the multiphoton microscope host 100, the laser light of different parameters emitted from the various lasers 200 can be adapted to the subsequent devices through adjustment of the laser adapter 300. Moreover, since the laser adapter 300 is connected with the multiphoton microscope host 100 through the transmission optical fiber 301, the multiphoton microscope 100 can freely move, and the multiphoton microscope host 100 can be placed at different positions or even in a cross-platform manner according to requirements, so that the use is more flexible.

The laser adapter 300 may specifically include a housing, and a beam conversion device and a beam stabilization device located in the housing, where the beam conversion device is configured to convert a laser beam entering the housing, and the beam conversion refers to amplifying, shrinking, zooming, and the like, by using conversion characteristics of an optical element, so that the laser can be matched with a subsequent device, and the best performance of the device is achieved. The beam stabilizing device is arranged at the downstream of the beam conversion device along the laser transmission direction and is used for adjusting the deflection direction of the laser beam so as to correct the deviation between the actual position of the laser beam at the laser output port and the ideal position. The beam stabilizing device is similar to the arrangement of the beam conversion device in the laser coupling module 3, and can adjust the deflection direction of laser when detecting that the laser beam deflects, so that the laser output is stable, and the coupling efficiency of the laser output is ensured.

In the embodiment shown in fig. 15, a fixed optical path may be provided between the laser 200 and the multiphoton microscope host 100, but in this case, there is no movement between the laser 200 and the multiphoton microscope host 100, and if the position is to be switched, the optical path needs to be re-set up.

The foregoing description of the preferred embodiments of the application is not intended to be limiting, but rather is to be construed as including any modifications, equivalents, and alternatives falling within the spirit and principles of the application.

Claims

1. The multi-photon microscope host machine is used for being connected with a microscope probe and is characterized by comprising an installation main body, and a wide-field searching module, a laser coupling module, a fluorescence collecting module, a scanning control module, a visual field searching adapter and a moving module which are integrated on the installation main body; wherein,

The fluorescence collection module is connected with the microscope probe through a fluorescence collection optical fiber and is used for collecting fluorescence signals output by the microscope probe;

The visual field searching adapter comprises a probe mounting assembly and a switching mechanism, wherein the probe mounting assembly is used for detachably mounting the microscope probe, the switching mechanism is arranged to be capable of switching the probe mounting assembly to a first position and a second position, when the probe mounting assembly is positioned at the first position, the microscope probe mounted on the probe mounting assembly avoids a light path between the wide field searching module and the living body, and when the probe mounting assembly is positioned at the second position, the microscope probe is aligned with the light path of the wide field searching module; and

The moving module is used for bearing a living body and can drive the living body to move in multiple directions, and the wide-field searching module is used for searching the field of view of the living body on the moving module.

2. The multiphoton microscope host of claim 1, further comprising a switchable light blocking door mounted on the mounting body;

3. The multiphoton microscope host of claim 1, wherein the mounting body comprises a base and a mounting bracket fixed to the base, the mounting bracket being provided with a support plate at an upper position thereof; wherein,

4. A multiphoton microscope host according to claim 2 or claim 3, wherein the wide field search module is mounted on the laser coupling module, the laser coupling module is provided with a light path through hole penetrating up and down, and a light path of the wide field search module is arranged to pass down through the light path through hole to reach the moving module.

5. The multi-photon microscope host computer of claim 1, wherein the laser coupling module comprises a coupler housing, a dispersion compensation element, an acousto-optic modulator and a beam stabilization device, wherein the dispersion compensation element, the acousto-optic modulator and the beam stabilization device are all arranged in the coupler housing and are sequentially arranged along a transmission direction of laser light; wherein,

the acousto-optic modulator is used for adjusting the intensity of laser;

6. The multiphoton microscope host of claim 5, wherein the laser coupling module further comprises a driver for driving the acousto-optic modulator and a cooling mechanism for cooling the driver, the driver and the cooling mechanism both being located on an upper surface of the coupler housing.

7. The multi-photon microscope host computer according to claim 6, wherein the wide field searching module is mounted on the upper surface of the coupler housing, the laser coupling module is provided with a light path through hole penetrating up and down, and the light path of the wide field searching module is arranged to penetrate down through the light path through hole;

8. The multiphoton microscope host of claim 1 or 2, further comprising a control box mounted on the mounting body, the fluorescence collection module and the scanning control module both being located within the control box.

9. The multiphoton microscope host of claim 8, wherein the control box is provided with a first interface for connecting the fluorescence collection optical fiber and a second interface for connecting the control cable, and the first interface and the second interface are both positioned above the same side of the control box;

10. The multiphoton microscope host of claim 9, further comprising a receiving device mounted on a side of the control box having the first interface and the second interface;

11. A multiphoton microscope system, the multiphoton microscope system comprising: a laser, a microscope probe and a multiphoton microscope host according to any one of claims 1 to 10;