CN109656014B

CN109656014B - Multipath fluorescence collection device and three-dimensional nonlinear laser scanning cavity endoscope

Info

Publication number: CN109656014B
Application number: CN201910099436.3A
Authority: CN
Inventors: 吴润龙; 王爱民; 江文茂; 胡炎辉; 程和平
Original assignee: Beijing Chaoweijing Biological Technology Co ltd
Current assignee: Beijing Chaoweijing Biological Technology Co ltd
Priority date: 2019-01-31
Filing date: 2019-01-31
Publication date: 2024-03-19
Anticipated expiration: 2039-01-31
Also published as: CN109656014A

Abstract

The embodiment of the invention provides a multipath fluorescence collection device and a three-dimensional nonlinear laser scanning cavity endoscope. The multi-channel fluorescence collection device comprises a dual-channel fluorescence collection module, an illumination multiplexing module and an imaging multiplexing module, wherein the dual-channel fluorescence collection module synchronously converges two-photon fluorescence signals and second harmonic signals collected by the illumination multiplexing module and the imaging multiplexing module, and then the two-photon fluorescence signals and the second harmonic signals are converted into corresponding electric signals. The multipath fluorescence collection device and the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention adopt modularized combination, collect two-photon fluorescence signals and second harmonic signals in front of the remote endoscope detection device through the optical fiber multiplexing function in the illumination multiplexing module and the imaging multiplexing module, and synchronously gather the two-photon fluorescence signals and the second harmonic signals to the two-channel fluorescence collection module, so that the two-channel fluorescence collection module collects most of the two-photon fluorescence signals and the second harmonic signals, and more accurate imaging of the cell structure is realized.

Description

Multipath fluorescence collection device and three-dimensional nonlinear laser scanning cavity endoscope

Technical Field

The embodiment of the invention relates to the technical field of laser scanning endoscopes, in particular to a multipath fluorescence collection device and a three-dimensional nonlinear laser scanning cavity endoscope.

Background

Gastrointestinal malignancy is the second leading cause of cancer death in the developed world population, and this trend has become more apparent in recent years. Surgical radical excision is mainly adopted for treating malignant tumor of gastrointestinal tract, but the specific range of the surgical excision needs to be determined when the surgical radical excision is carried out, so that the benign and malignant tumor, the infiltration depth, the metastasis condition, the existence of cancer residues at the incisional edge and the like need to be known before the operation is carried out. Therefore, preoperative gastrointestinal biopsy is an important diagnostic evidence for histological diagnosis of gastrointestinal tumors. And according to tumor size, growth position, infiltration depth, etc., gastric cancer is classified into gastric total incision, gastric sub-total incision, partial gastrectomy, endoscopic submucosal or submucosal resection, etc.

At present, a gastrointestinal endoscope which is used for detecting and imaging the cell structures of gastrointestinal tissues in the abdominal cavity, the oral cavity and the intrauterine tissues of a human body is provided with a cavity endoscope based on a two-photon imaging technology, but most of the gastrointestinal endoscopes collect two-photon signals through an objective lens in a detection device and then carry out single light path processing through a fluorescence collection device at a background.

The fluorescence collection device only integrates a single two-photon processing light path, has a single function, and cannot meet the requirement of collecting fluorescence signals in multiple paths.

Disclosure of Invention

Aiming at the technical problems in the prior art, the embodiment of the invention provides a multipath fluorescence collection device and a three-dimensional nonlinear laser scanning cavity endoscope.

In a first aspect, an embodiment of the present invention provides a multiple fluorescence collection device, including:

the system comprises a dual-channel fluorescence collection module, an illumination multiplexing module and an imaging multiplexing module, wherein the illumination multiplexing module and the imaging multiplexing module are both in optical fiber communication connection with the dual-channel fluorescence collection module, and the dual-channel fluorescence collection module comprises the following components:

the illumination multiplexing module is used for providing illumination light signals for the remote endoscope detection device, and transmitting the two-photon fluorescence signals and the second harmonic signals to the dual-channel fluorescence collection module after collecting the two-photon fluorescence signals and the second harmonic signals based on an illumination optical fiber bundle;

the imaging multiplexing module is used for imaging a tissue region to be detected in front of an objective lens of the remote endoscope detection device, and transmitting the two-photon fluorescence signal and the second harmonic signal to the dual-channel fluorescence collection module after collecting the two-photon fluorescence signal and the second harmonic signal based on a bright field optical fiber bundle;

the dual-channel fluorescence collection module is used for collecting two-photon fluorescence signals and second harmonic signals based on an objective lens in the remote endoscope detection device, synchronously converging the two-photon fluorescence signals and the second harmonic signals transmitted by the illumination multiplexing module and the imaging multiplexing module, and then converting the two-photon fluorescence signals and the second harmonic signals into corresponding electric signals; .

In a second aspect, embodiments of the present invention provide a three-dimensional nonlinear laser scanning cavity endoscope, comprising:

the device comprises a cavity endoscope detection device, a scanning acquisition controller, a femtosecond pulse laser, an optical fiber coupling module and the multipath fluorescence collection device provided by the first aspect of the embodiment of the invention, wherein the multipath fluorescence collection device and the optical fiber coupling module are connected with the zoom type cavity endoscope detection device in an optical fiber communication mode, and the multipath fluorescence collection device and the cavity endoscope detection device are electrically connected with the scanning acquisition controller, wherein:

the femtosecond pulse laser is used for outputting pulse laser signals to the optical fiber coupling module;

the optical fiber coupling module is used for coupling the pulse laser signals output by the femtosecond pulse laser and transmitting the pulse laser signals to a collimating lens in the cavity endoscope detection device;

the cavity endoscope detection device is used for receiving the pulse laser signals, outputting the pulse laser signals to autofluorescent substances in living cells, acquiring fluorescent signals and second harmonic signals generated after the autofluorescent substances are excited, and outputting the fluorescent signals and the second harmonic signals to the fluorescent collection device;

The scanning acquisition controller is used for controlling the micro-electromechanical scanning galvanometer in the cavity endoscope detection device to scan the pulse laser signal and synchronously acquiring the electric signal.

The multipath fluorescence collection device and the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention adopt modularized combination, collect two-photon fluorescence signals and the second harmonic signals in front of the far-end endoscope detection device through the optical fiber multiplexing function in the illumination multiplexing module and the imaging multiplexing module, and synchronously gather the two-photon fluorescence signals and the second harmonic signals to the double-channel fluorescence collection module, so that the double-channel fluorescence collection module can collect most of the two-photon fluorescence signals and the second harmonic signals in front of the far-end endoscope detection device, and more accurate cell structure imaging is realized.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic diagram of a multi-path fluorescence collection device according to an embodiment of the present invention;

FIG. 2 is a schematic diagram of the dual-channel fluorescence collection module in the multi-channel fluorescence collection device according to the embodiment of the present invention;

FIG. 3 is a schematic view of a three-dimensional nonlinear laser scanning cavity endoscope according to an embodiment of the present invention;

FIG. 4 is a schematic view of a three-dimensional nonlinear laser scanning cavity endoscope according to another embodiment of the present invention;

FIG. 5 is a schematic view of a cavity endoscopic probe apparatus according to an embodiment of the present invention;

FIG. 6 is a schematic view of a endoscopic probe apparatus according to another embodiment of the present invention;

FIG. 7 is a schematic view of a three-dimensional nonlinear laser scanning cavity endoscope according to still another embodiment of the present invention;

FIG. 8 is a schematic diagram of a box-type combined structure of a three-dimensional nonlinear laser scanning cavity endoscope according to an embodiment of the present invention;

FIG. 9 is a schematic diagram of a box-type combined structure of a three-dimensional nonlinear laser scanning cavity endoscope according to another embodiment of the present invention;

FIG. 10 is a schematic view of a table structure of a three-dimensional nonlinear laser scanning cavity endoscope according to an embodiment of the present invention;

FIG. 11 is a schematic view of a desktop structure of a three-dimensional nonlinear laser scanning cavity endoscope, according to another embodiment of the present invention.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

At present, imaging is carried out based on a gastroenteroscope by taking CT, MRI and the like as assistance to obtain relevant information such as benign and malignant tumor, infiltration depth, metastasis condition, existence of cancer residues at a cutting edge and the like, and the imaging method has the defects in specific operation, such as easy bleeding of an intestinal canal or a tumor body, need of manual traction or extrusion, delay caused by repeated endoscopic biopsy when the gastroenteroscope cannot pass through the intestinal canal, and need of additional first aid hemostasis if severe bleeding is caused. However, the auxiliary examination means such as CT and MRI cannot accurately judge the infiltration depth of early gastrointestinal tumor and the lymph node metastasis in clinical practice. The ultrasonic endoscope is used for judging the T stage of the gastrointestinal tumor, the literature reports that the accuracy is only 44.7% -78%, the ultrasonic endoscope is insufficient to become a reliable diagnosis standard, the ultrasonic endoscope has poor preoperative judgment effect on the partial excision operation, the gastrointestinal mucosa layer cannot be accurately subdivided, and the N stage effect is poor.

Whereas conventional white light laparoscopes and endoscopes are capable of assessing many gastrointestinal disorders, the technique is limited to detecting gross morphological changes. Although suspicious regions are easily found, these techniques are related to false positive rate, specificity, and the like, as compared to in vivo detection techniques. White light endoscopy is associated with extensive errors in microscopic change diagnosis, including the diagnosis of ulcerative colitis or Barrett's oesophagus and flat adenomatous dysplasia. Confocal endoscopes have received extensive attention in combination with laser technology, fluorescence detection technology, rapid scanning technology, and the like, because of their ability to detect mucosal changes at the microscopic level, and the potential for replacement of tissue biopsies, the imaging technology has high sensitivity and specificity. However, the confocal endoscopic imaging technology is still limited by imaging depth and fluorescent dye, and because the gastrointestinal sample has strong absorption and scattering to visible light, the imaging depth is only in a shallow surface layer, and a specific fluorescent dye developer is also required to be injected, the operation is too complicated, and the relevant information such as the infiltration depth of tumor, the metastasis condition, the existence of cancer residues at the surgical incision edge and the like can not be accurately obtained.

The two-photon microscopic imaging technology adopts a femtosecond pulse laser with longer wavelength as an excitation light source, has the characteristics of deep imaging depth, small photodamage, small photobleaching area, high fluorescence collection efficiency and the like, and has epoch-making significance in deep imaging of biological tissues. W.Denk et al, university of Conneler, 1990 developed the first two-photon fluorescence microscope in the world, using multiphoton microscopy imaging techniques based on nonlinear optics and femtosecond pulsed lasers. The technology can rapidly obtain the tissue structure and the cell morphology of the specimen in real time by utilizing the self-fluorescence generated by cells in living tissues and the second harmonic generated by collagen tissues. As early as 1986, the second harmonic was used in skin studies and in coronary microscopy imaging studies, confirming its feasibility to be used for the observation of biological tissues. MPM may also be an important tool in cancer research. The autofluorescence generated by the cells is derived from Nicotinamide Adenine Dinucleotide (NADH) and Flavin Adenine Dinucleotide (FAD) in the cells, the wavelength of NADH is 460nm, and the secondary oscillation harmonic wave of collagen is 370-390 nm, so a multiphoton microscope in the range of 780-940 nm is usually selected when observing tumor specimen tissues. MPM imaging is not only comparable to standard tumor histopathology, but also provides additional information on tumor neogenesis processes, as reflected in the metabolic levels of tumor tissue cells by measuring the ratio NADH/FAD.

Using multiphoton imaging techniques, multiphoton microscopes can provide real-time gastrointestinal tissue structure and cell morphology information. The multiphoton imaging technology has the characteristics of no need of externally marking tissues, extremely sensitivity to collagen, small photodamage to tissues, deep penetration depth and the like, and can be applied to optical biopsy of gastrointestinal tumors. There are no clinically available two-photon laparoscope, endoscope and cavity endoscope detection device based on two-photon imaging to detect gastrointestinal tissue information in situ in real time.

Currently, there are gastrointestinal endoscopes which are used for detecting and imaging the cell structures of gastrointestinal tissues, oral cavity and intrauterine tissues in the abdominal cavity of a human body, and cavity endoscopes based on the two-photon imaging technology, but most of them collect two-photon signals through an objective lens in a detecting device and then carry out single light path processing through a fluorescence collecting device at the background. The fluorescence collection device only integrates a single two-photon processing light path, has a single function, and cannot meet the requirement of collecting fluorescence signals in multiple paths.

In order to realize multi-path collection of fluorescent signals, an embodiment of the present invention provides a multi-path fluorescent collection device, and fig. 1 is a schematic structural diagram of the multi-path fluorescent collection device provided by the embodiment of the present invention, as shown in fig. 1, where the device includes:

The dual-channel fluorescence collection module 11, the illumination multiplexing module 12 and the imaging multiplexing module 13 are all connected with the dual-channel fluorescence collection module 11 in an optical fiber communication mode, wherein:

the illumination multiplexing module 12 is configured to provide an illumination light signal for the remote endoscope detection device, and transmit the two-photon fluorescence signal and the second harmonic signal to the dual-channel fluorescence collection module after collecting the two-photon fluorescence signal and the second harmonic signal based on an illumination fiber bundle;

the imaging multiplexing module 13 is configured to image a tissue region to be detected in front of an objective lens of the distal endoscope detection device, collect two-photon fluorescence signals and second harmonic signals based on a bright field optical fiber bundle, and then transmit the two-photon fluorescence signals and the second harmonic signals to the dual-channel fluorescence collection module;

the dual-channel fluorescence collection module 11 is configured to collect two-photon fluorescence signals and second harmonic signals based on an objective lens in a remote endoscope detection device, and convert the two-photon fluorescence signals and the second harmonic signals into corresponding electrical signals after synchronously converging the two-photon fluorescence signals and the second harmonic signals transmitted by the illumination multiplexing module and the imaging multiplexing module; .

Specifically, the multi-path fluorescence collection device provided by the embodiment of the invention mainly comprises three modules, namely, a dual-path fluorescence collection module 11, an illumination multiplexing module 12 and an imaging multiplexing module 13, wherein the illumination multiplexing module 12 and the imaging multiplexing module 13 are respectively in communication connection with optical fibers of the dual-path fluorescence collection module 11, the illumination multiplexing module 12 and the imaging multiplexing module 13 can collect partial fluorescence signals including two-photon fluorescence signals and second harmonic signals, meanwhile, the illumination multiplexing module 12 also has the function of providing illumination optical signals for a far-end endoscope detection device, the imaging multiplexing module 13 also has the function of imaging a tissue region to be detected in front of an objective lens of the far-end endoscope detection device, the illumination multiplexing module 12 and the imaging multiplexing module 13 transmit the collected two-photon fluorescence signals and second harmonic signals to the fluorescence collection module 11 through optical fibers, the dual-path fluorescence collection module 11 collects two-photon fluorescence signals and second harmonic signals through the objective lens in the far-end endoscope detection device, and synchronously converges the two-photon fluorescence signals and the second harmonic signals collected by the illumination multiplexing module 12 and the imaging multiplexing module 13, the two-photon fluorescence signals and the second harmonic signals are converted into corresponding electrical signals, the two-photon fluorescence signals are collected by the two-path fluorescence collection module 11 through the two-path fluorescence collection module and the two-path fluorescence collection module 11, the two-channel fluorescence collection module 11 gathers the second harmonic signals transmitted by the illumination multiplexing module 12 and the imaging multiplexing module 13 and the second harmonic signals acquired by the object lens into another path, and converts the gathered second harmonic signals into second electric signals, wherein the illumination multiplexing module is in optical fiber communication connection with the far-end endoscope detection device through an illumination optical fiber bundle, provides illumination optical signals for the far-end endoscope detection device, and synchronously collects two-photon fluorescence signals and second harmonic signals before the far-end endoscope detection device; the imaging multiplexing module is in optical fiber communication connection with the far-end endoscope detection device through the bright field optical fiber bundle, images the tissue region to be detected in front of the objective lens of the far-end endoscope detection device, and synchronously collects two-photon fluorescence signals and second harmonic signals in front of the far-end endoscope detection device.

The multi-path fluorescence collection device provided by the embodiment of the invention adopts modularized combination, acquires the two-photon fluorescence signals and the second harmonic signals in front of the remote endoscope detection device through the optical fiber multiplexing function in the illumination multiplexing module and the imaging multiplexing module, and synchronously gathers the two-photon fluorescence signals and the second harmonic signals to the dual-channel fluorescence collection module, so that the multi-path fluorescence collection device can collect more two-photon fluorescence signals and the second harmonic signals in front of the remote endoscope detection device relative to the existing fluorescence collection device through the object lens collecting optical fiber single-path collecting two-photon fluorescence signals and the second harmonic signals in front of the remote endoscope detection device, and converts the two-photon fluorescence signals and the second harmonic signals to corresponding electric signals through the dual-channel fluorescence collection module, thereby realizing more accurate cell structure imaging.

On the basis of the foregoing embodiments, the illumination multiplexing module in the multiple fluorescence collection device provided by the embodiment of the present invention includes an illumination light path and a first multiplexing collection light path, as shown in fig. 1, where:

the illuminated light path includes, in order, an illumination fiber bundle 121, a first multiplexed dichroic mirror 122, a variable filter 123, an illumination lens 124, and an illumination light source 125;

the first multiplexed collection optical path includes, in order, an illumination fiber bundle 121, a first multiplexed dichroic mirror 122, a first multiplexed collection lens 126, and a first transmission fiber 127. Namely, the illumination multiplexing module in the multiplex fluorescence collection device provided by the embodiment of the invention has illumination and fluorescence collection functions, the corresponding light paths are an illumination light path and a first multiplex collection light path, wherein the illumination light path comprises an illumination fiber bundle 121, a first multiplex dichroic mirror 122, a variable optical filter 123, an illumination lens 124 and an illumination light source 125, the illumination light source 125 emits illumination light signals to the illumination lens 124, the illumination light signals are converged to the variable optical filter 123, after being filtered, the illumination light signals are transmitted to the first multiplex dichroic mirror 122 and are transmitted to a far-end endoscope detection device through the illumination fiber bundle 121, illumination is provided for the endoscope detection device, and meanwhile, the first multiplex collection light path sequentially comprises the illumination fiber bundle 121, the first multiplex dichroic mirror 122, the first multiplex collection lens 126 and the first transmission fiber 127, the illumination fiber bundle 121 collects two-photon fluorescence signals and the second harmonic signals before the far-end endoscope detection device and transmits the two-photon fluorescence signals and the second harmonic signals to the first multiplexing dichroic mirror 122, the two-photon fluorescence signals and the second harmonic signals are reflected to the first multiplexing collecting lens 126 through the first multiplexing dichroic mirror 122, the converging and beam-combining are transmitted to the first transmission fiber 127, the first transmission fiber 127 is transmitted to the dual-channel fluorescence collecting module, and fluorescence collecting efficiency is improved, so that achieve the purpose of improving the image signal-to-noise ratio is achieved, wherein the illumination light source 125 passes through an electric variable optical filter 123 runner, different optical filters can be switched to obtain illumination light signals with different wavelengths, the basic principle is that the two-photon fluorescence imaging is not interfered, for example, when the two-photon fluorescence imaging is achieved, the two-photon fluorescence imaging light source 125 can be switched to red or infrared light signals with the second harmonic imaging is achieved, the illumination light signals with the fluorescence light signals with the wavelength of 370nm, the 635nm or the infrared light signals with the wavelength are obtained, and the illumination light signals are coupled into the illumination fiber bundle 121 through a lens.

On the basis of the foregoing embodiments, the imaging multiplexing module in the multiplex fluorescence collection device provided by the embodiment of the present invention includes an imaging optical path and a second multiplexing collection optical path, as shown in fig. 1, where:

the imaging light path sequentially comprises a bright-field optical fiber bundle 131, a second multiplexing dichroic mirror 132, an imaging lens 133 and a camera 134;

the second multiplexing collection optical path includes, in order, a bright field optical fiber bundle 131, a second multiplexing dichroic mirror 132, a second multiplexing collection lens 135, and a second transmission optical fiber 136. Namely, the imaging multiplexing module in the multi-path fluorescence collection device provided by the embodiment of the invention has the functions of imaging and fluorescence collection, the corresponding optical paths are an imaging optical path and a second multiplexing collection optical path, wherein the imaging optical path sequentially comprises a bright field optical fiber bundle 131, a second multiplexing dichroic mirror 132, an imaging lens 133 and a camera 134, the camera 134 captures the information of the tissue region to be detected before the objective lens of the far-end endoscope detection device through the bright field optical fiber bundle 131, the bright field optical fiber bundle 131 collects the two-photon fluorescence signal before the far-end endoscope detection device and the second harmonic signal through the optical fiber multiplexing function of the bright field optical fiber bundle 131, the two-photon fluorescence signal and the second harmonic signal are transmitted to the second multiplexing dichroic mirror 132, the two-photon fluorescence signal is reflected to the second multiplexing collection lens 135 through the second multiplexing dichroic mirror 132, the two-photon fluorescence collection module is converged to the second transmission optical fiber 136, the camera 134 can be two laparoscopes corresponding to the two binocular bright field optical fiber bundles 131, the bright field imaging and the two-photon imaging lens form a multi-mode mirror, the bright field binocular three-dimensional stereoscopic mode is used for carrying out the basic observation of the sample with large visual field, and the appearance is observed. For suspicious or interested areas, the two-photon mode can be switched to perform autofluorescence and second harmonic imaging, and the cell-level morphology of the sample is observed, so that basis is provided for further judgment, wherein the camera 134 can be an imaging device based on CCD or CMOS imaging devices.

On the basis of the above embodiments, fig. 2 is a schematic structural diagram of the dual-channel fluorescence collection module in the multi-channel fluorescence collection device according to the embodiment of the present invention, as shown in fig. 2, the dual-channel fluorescence collection module in the multi-channel fluorescence collection device according to the embodiment of the present invention includes an objective lens collection optical fiber, an optical fiber universal interface 881, a first photomultiplier 882, a second photomultiplier 883, a first collection optical path located between the optical fiber universal interface 881 and the first photomultiplier 882, and a second collection optical path located between the optical fiber universal interface 881 and the second photomultiplier 883, wherein:

the first transmission optical fiber, the second transmission optical fiber and the objective lens collection optical fiber are all in communication connection with the optical fiber universal interface optical fiber;

the first collecting light path sequentially comprises a coupling lens 81, an infrared filter 82, a first dichroic mirror 83, a first filter 84 and a first collecting lens 85, wherein the first collecting light path is used for collecting fluorescent signals received by the fluorescent collecting device, and the first photomultiplier 882 is used for converting the fluorescent signals into first electric signals;

the second collecting light path sequentially includes a coupling lens 81, an infrared filter 82, a first dichroic mirror 83, a second dichroic mirror 86, a second filter 87, and a second collecting lens 88, where the second collecting light path is used to collect the second harmonic signal received by the fluorescent collecting device, and the second photomultiplier 883 is used to convert the second harmonic signal into a second electric signal. The two-channel fluorescence collection module in the multi-channel fluorescence collection device provided by the embodiment of the invention integrates two paths of signal collection light paths, namely a fluorescence signal collection light path and a second harmonic signal collection light path, respectively, and two-photon fluorescence signals collected by the illumination multiplexing module, the imaging multiplexing module and the two-channel fluorescence collection module are respectively collected and converted into first electric signals and second electric signals, and the second harmonic signals collected by the illumination multiplexing module, the imaging multiplexing module and the two-channel fluorescence collection module are processed by the two-channel fluorescence collection module and the second harmonic signals, wherein the processed fluorescence signals and the second harmonic signals come from fluorescence signals and second harmonic signals collected by an objective lens in a remote endoscope detection device, and the illumination multiplexing module and the imaging multiplexing module are converged into the fluorescence signals and the second harmonic signals in the two-channel fluorescence collection module.

The first dichroic mirror 83 in the first collecting light path is a dichroic mirror that transmits the two-photon fluorescence signals collected by the illumination multiplexing module, the imaging multiplexing module and the dual-channel fluorescence collecting module, reflects the second harmonic collected by the illumination multiplexing module, the imaging multiplexing module and the dual-channel fluorescence collecting module, the second dichroic mirror 86 and the first dichroic mirror 83 are the same dichroic mirror for reflecting the second harmonic, the first filter 84 is used for transmitting the fluorescence signals and filtering out the rest interference signals, the second filter 87 is used for transmitting the corresponding second harmonic signals and filtering out the rest interference signals, for example, when the 780nm femtosecond optical fiber laser is used for exciting the autofluorescence substances in the abdominal cavity or oral cavity cells of a human body, 390nm second harmonic signals and 450-600nm two-photon autofluorescence signals can be obtained, two-way fluorescence can be separated by the dichroic mirror that reflects the wavelength of more than 420nm, the first filter 84 of 390 +/-20 nm and the second filter 87 of 450-600nm are respectively used for reflecting the second harmonic signals, wherein the second filter 87 is used for transmitting the rest interference signals, the second harmonic signals are collected by the optical fiber and the optical fiber is connected with the optical fiber for collecting the two-photon autofluorescence signals in the far-end internal detection device, and the optical fiber is connected to the optical fiber interface.

On the basis of the above embodiments, the multiple illumination multiplexing modules in the multiple fluorescence collection device provided by the embodiment of the invention are provided. That is, a plurality of illumination multiplexing modules can be arranged in the multipath fluorescence collection device provided by the embodiment of the invention so as to collect fluorescence signals and second harmonic signals to the maximum extent.

On the basis of the above embodiments, the imaging multiplexing modules in the multiplex fluorescence collection device provided by the embodiment of the invention are multiple. That is, a plurality of imaging multiplexing modules can be arranged in the multipath fluorescence collection device provided by the embodiment of the invention so as to collect fluorescence signals and second harmonic signals to the maximum extent.

On the basis of the above embodiments, as shown in fig. 1, the multi-path fluorescence collection device provided in the embodiment of the present invention further includes an industrial personal computer 14, the dual-path fluorescence collection module is connected to the industrial personal computer 14 through optical fiber communication, the illumination multiplexing module and the imaging multiplexing module are both electrically connected to the industrial personal computer, and the industrial personal computer 14 is configured to obtain a first electrical signal and a second electrical signal, generate a first fluorescence image based on the first electrical signal, and generate a second fluorescence image based on the second electrical signal. Namely, after the fluorescence signals and the second harmonic signals collected by the multipath fluorescence collection device provided by the embodiment of the invention rotate into the corresponding first electric signals and second electric signals, the corresponding first fluorescence images and second fluorescence images are obtained and generated by the industrial personal computer 14, the industrial personal computer generates the first fluorescence images based on the first electric signals and generates the second fluorescence images based on the second electric signals, and the first fluorescence images and the second fluorescence images can be respectively used for displaying cell structure information and fiber structure information, wherein control software is installed on the industrial personal computer, and a control instruction is sent to a scanner through the control software so as to control the scanning collection controller to obtain the first electric signals and the second electric signals.

The embodiment of the invention also provides a three-dimensional nonlinear laser scanning cavity endoscope, fig. 3 is a schematic structural diagram of the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention, and as shown in fig. 3, the three-dimensional nonlinear laser scanning cavity endoscope comprises:

the utility model provides a multichannel fluorescence collection device of cavity endoscope detection device 18, scanning collection controller 15, femtosecond pulse laser, fiber coupling module, and each embodiment of the aforesaid provides, multichannel fluorescence collection device with fiber coupling module all with cavity endoscope detection device 18 fiber communication connects, multichannel fluorescence collection device with cavity endoscope detection device 18 all with scanning collection controller 15 electricity is connected, wherein:

the optical fiber coupling module is used for coupling the pulse laser signals output by the femtosecond pulse laser and transmitting the pulse laser signals to a collimating lens in the cavity endoscope detection device 18;

the cavity endoscope detecting device 18 is configured to receive the pulse laser signal, output the pulse laser signal to an autofluorescent substance in a living cell, obtain a fluorescence signal and a second harmonic signal generated after excitation of the autofluorescent substance, and output the fluorescence signal and the second harmonic signal to the fluorescence collecting device;

The scanning acquisition controller 15 is configured to control a microelectromechanical scanning galvanometer in the endoscopic detection device 18 to scan the pulsed laser signal, and to synchronously acquire the electrical signal.

Specifically, the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention comprises a cavity endoscope detection device 18, a scanning acquisition controller 15, a femtosecond pulse laser, an optical fiber coupling module and the multipath fluorescence collection device provided by the above embodiments, so as to form the three-dimensional nonlinear laser scanning cavity endoscope for detecting gastrointestinal tissues and oral tissues of a human body by utilizing a two-photon imaging technology, wherein the femtosecond pulse laser can emit pulse laser signals for exciting autofluorescent substances in gastrointestinal tissues and oral tissue cells of the human body to generate multiphoton fluorescence signals and second harmonic signals, the femtosecond pulse laser of 920nm is used for exciting FAD and collagen in cells, the fluorescence signals of 500-600nm and the second harmonic signals of 460nm are excited, and the autofluorescent substances such as FAD or NADH in cells are excited by the femtosecond pulse laser of 780nm to generate corresponding fluorescence signals and second harmonic signals, wherein the femtosecond pulse laser and the optical fiber coupling module are combined together to form a laser emission module;

The multi-channel fluorescence collection device comprises a dual-channel fluorescence collection module 11, an illumination multiplexing module 12 and an imaging multiplexing module 13, two paths of signal collection light paths are integrated, and the two paths of signal collection light paths and a second harmonic signal collection light path are respectively used for realizing the respective collection of fluorescence signals and second harmonic signals; the scanning acquisition controller 15 controls the micro-electromechanical scanning galvanometer to scan the pulse laser signal and excite the autofluorescent substance to generate a fluorescent signal and a second harmonic signal, and acquires a first electric signal and a second electric signal obtained by converting the fluorescent signal and the second harmonic signal by the fluorescent collection device; the three-dimensional nonlinear laser scanning cavity endoscope can be divided into a laparoscope and a stomatoscope according to the different structures of the cavity endoscope detection device 18, as shown in fig. 3, and if the cavity endoscope detection device 18 is a laparoscope detection device, the three-dimensional nonlinear laser scanning cavity endoscope is a laparoscope. Fig. 4 is a schematic structural diagram of a three-dimensional nonlinear laser scanning cavity endoscope according to another embodiment of the present invention, and as shown in fig. 4, the cavity endoscope detection device 18 is an stomatoscope detection device, and then the three-dimensional nonlinear laser scanning cavity endoscope is an stomatoscope. The resolution of the three-dimensional nonlinear laser scanning cavity endoscope can be set to 800nm, the imaging field of view can be 400 micrometers by 400 micrometers, and the imaging speed can be 26 frames (256×256 pixels) or 13 frames (512×512 pixels).

The three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention adopts the multipath fluorescence collection device, the scanning acquisition controller, the femtosecond pulse laser, the optical fiber coupling module and the cavity endoscope detection device, so that the laser scanning cavity endoscope for detecting gastrointestinal tissues and oral tissues of a human body by utilizing a two-photon imaging technology is formed, and the multipath fluorescence collection device is used for multipath collection of fluorescence signals and second harmonic signals generated after front cells of the cavity endoscope detection device are excited by adopting the multipath fluorescence collection module, the illumination multiplexing module and the imaging multiplexing module, so that corresponding cell tissue structure information is obtained, and the device is simple to operate and convenient to use.

On the basis of the above embodiments, the cavity endoscope detecting device in the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the present invention includes an outer fixed housing and an inner clamping device, fig. 5 is a schematic structural diagram of the cavity endoscope detecting device provided by the embodiment of the present invention, as shown in fig. 5, the inner clamping device 182 is disposed in the outer fixed housing 181, a zoom motor is disposed inside the outer fixed housing 181 and is used for driving the inner clamping device 182 to move up and down relative to the outer fixed housing 181, and an optical path structure for forming a first optical path and a second optical path is disposed in the inner clamping device 182, where:

The first optical path sequentially includes a collimating lens 1821, a mems scanning galvanometer 1822, a lens 1823, a dichroic mirror 1824, a relay lens 1825, and an objective lens 1826, where the first optical path is used for conducting the laser signal received by the collimating lens 1821 from the collimating lens 1821 to the objective lens 1826;

the second optical path includes the objective lens 1826, the relay lens 1825, and the dichroic mirror 1824 in order, where the second optical path is used to conduct the optical signal collected by the objective lens 1826 from the objective lens 1826 to the dichroic mirror 1824. Namely, the endoscope detection device in the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention comprises an outer fixed shell 181 and an inner clamping device 182, wherein the inner clamping device 182 is an integral body, optical paths for transmitting laser signals and collecting two-photon signals and second harmonic signals are arranged in the inner clamping device 182, the optical paths are respectively a first optical path and a second optical path, the first optical path comprises a collimating lens 1821, a micro-electromechanical scanning galvanometer 1822, a lens, a dichroic mirror, a relay mirror 1825 and an objective lens 1826, the laser signals for exciting autofluorescent substances in human gastrointestinal tissues or oral tissue cells are emitted through the collimating lens 1821, the micro-electromechanical scanning galvanometer 1822, the lens, the dichroic mirror, the relay mirror 1825 and the objective lens 1826 in the first optical path, the autofluorescent substances are emitted from the objective lens 1826 to the autofluorescent substances, the two-photon signals and the second harmonic signals are collected through the objective lens 1826, the two-photon signals and the second harmonic signals are collected through the relay mirror 1825 and the dichroic mirror in the second optical path, the two-photon signals are collected into the fluorescent collecting device, the detection information for acquiring the gastrointestinal tissues or oral tissue infiltration detection information is acquired, the information is judged by the aid of the motor, and the relative depth of the optical path is different from the optical paths, the optical paths are not fixed with the optical paths, and the optical paths are not fixed with the optical path structures, and the optical paths are not clamped by the device.

Wherein a relay lens 1825 is disposed inside the objective lens 1826, and is used for conducting the laser signal for exciting the autofluorescent substance from the dichroic mirror to the objective lens 1826 for a long distance, and conducting the two-photon signal and the second harmonic signal collected by the objective lens 1826 to the dichroic mirror, wherein the image plane of the laser signal objective lens 1826 coincides with the focal plane of the relay lens 1825, and the laser signal scanning area passing through the micro-electromechanical scanning mirror is conducted to the image plane of the objective lens 1826 in a ratio of 1:1, wherein the relay lens 1825 can be extended or shortened according to specific needs.

The dichroic mirror 1824 may be configured as a long-pass short-reflecting dichroic mirror or a short-pass long-reflecting dichroic mirror, that is, when the long-pass short-reflecting dichroic mirror is configured, a pulse laser signal for exciting an autofluorescent substance is transmitted, and the collected two-photon signal and the second harmonic signal are reflected, where the variable-focus cavity endoscope detection device may be a laparoscopic detection device; when the dichroic mirror 1824 is a short-pass long-reflecting dichroic mirror, a pulse laser signal for exciting an autofluorescent substance is reflected, the collected two-photon signal and a second harmonic signal are transmitted, the dichroic mirror reflects the laser signal which is incident on the dichroic mirror after passing through the collimating lens 1821, the micro-electromechanical scanning galvanometer 1822 and the lens, and passes through the relay lens 1825 to the objective lens 1826, and the two-photon signal and the second harmonic signal collected by the objective lens 1826 are transmitted, and at this time, the variable-focus cavity endoscope detection device can be a detection device of an oral cavity mirror.

On the basis of the above embodiments, the optical path structure in the endoscopic detection apparatus provided in the embodiment of the present invention further includes a liquid lens, and fig. 6 is a schematic structural diagram of an endoscopic detection apparatus provided in another embodiment of the present invention, as shown in fig. 6, the liquid lens 1820 is located between the collimating lens 1821 and the mems scanning galvanometer 1822, so as to form a new first optical path, where the new first optical path includes the collimating lens 1821, the liquid lens 1820, the mems scanning galvanometer 1822, the lens 1823, the dichroic mirror 1824, and the objective lens 1826 in sequence. That is, the liquid lens 1820 is arranged such that the liquid lens 1820 is bent by applying a voltage or a current to the liquid lens 1820, so that the parallel light emitted from the straight lens 1821 has different optical powers. The specific light path is: the laser signal is emitted from the optical fiber, is parallel to enter the liquid lens 1820 after passing through the collimating lens 1821, generates corresponding optical power according to the loaded voltage or current signal from the liquid lens 1820, and the emitted converging or diverging light is transmitted to the objective lens 1826 through the relay lens 1825 and then converged on the sample after passing through the micro-electromechanical scanning galvanometer 1822, the lens 1823 and the dichroic mirror 1824. The focal power change introduced by the liquid lens 1820 can enable the focal point of the laser signal emitted from the opening of the objective lens 1826 to move back and forth in the depth direction, the response speed of the liquid lens 1820 is very fast, and the scanning frequency is in the order of KHz, so that fast scanning imaging in the depth direction can be realized. Wherein the liquid lens 1820 is equivalent to a parallel plate glass when no voltage or current signal is applied, has no optical power to the laser signal and does not cause any shift of the focus after the objective lens 1826, thereby realizing three-dimensional stereoscopic imaging. In specific use, the liquid lens 1820 is complementary to the zoom motor 183, the position of the objective lens 1826 is adjusted by the zoom motor 183, after coarse adjustment to the corresponding depth position, the system is switched to the liquid lens 1820 zoom scanning mode, and rapid three-dimensional imaging is performed on the sample, wherein when the zoom motor 183 is not installed in the variable-focus cavity endoscopic detection device, zoom adjustment can be performed only by the liquid lens 1820.

On the basis of the above embodiments, the outer fixing housing in the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the present invention includes a handle housing and a probe tube, as shown in fig. 5, the handle housing 1811 is fixedly connected with the probe tube 1812, the zoom motor is disposed inside the handle housing 1811, and a probe channel is disposed in the probe tube 1812, where:

the collimating lens 1821, the mems scanning galvanometer 1822, the lens, and the dichroic mirror in the optical path structure are all located in the handle housing 1811, the relay lens and the objective lens in the optical path structure are all located in the detection channel, and the objective lens is located at a channel port of the detection channel;

a plurality of illumination channels 1022 are further disposed in the detection tube 1812, and illumination optical fiber bundles for transmitting illumination light signals are disposed in the illumination channels 1022, wherein the illumination channels 1022 are uniformly distributed with the axis of the detection channel as the center;

an observation channel is further disposed in the probe tube 1812, the observation channel is located between the probe channel and the illumination channel 1022, an observation lens is disposed at a channel opening of the observation channel, and the observation lens is connected with a bright field optical fiber bundle in the observation channel, so as to obtain image information of a tissue region to be measured in front of the objective lens. That is, a plurality of illumination channels 1022 are further disposed in the probe tube 1812 in the endoscope probe device provided by the embodiment of the present invention, and an illumination fiber bundle for transmitting illumination light signals is disposed in the illumination channels 1022, where the illumination channels 1022 are uniformly distributed with the axis of the probe channel as the center. That is, a plurality of illumination channels 1022 are further disposed in the probe tube 1812 of the endoscope probe device provided in the embodiment of the present invention, more than one illumination channel 1022 is disposed in each channel, the illumination fibers have a certain aperture angle, the illumination channels 1022 can be directly used for divergent illumination without lenses, and the illumination channels 1022 are uniformly distributed with the axis of the probe channel as the center, so as to provide uniform illumination for the endoscope probe device, so as to facilitate working observation of the state of the tissue area to be measured in front of the objective lens.

An observation channel is further arranged in a detection tube of the cavity endoscope detection device, the observation channel is located between the detection channel and the illumination channel, an observation lens 1021 and a bright field optical fiber bundle are arranged, the bright field optical fiber bundle is an imaging optical fiber bundle and is used for transmitting image information of a tissue region to be detected before an objective lens captured by the observation lens, one observation channel or two observation channels can be used for forming binocular observation, a stereoscopic bright field cavity endoscope function is achieved, the binocular observation is shown in fig. 4, the three-dimensional nonlinear laser scanning cavity endoscope structure schematic diagram is shown in fig. 7, the imaging multiplexing module 13 is one, and the corresponding observation lens 1021 is one, namely the monocular observation.

On the basis of the above embodiments, an adsorption channel is further disposed in the detection tube in the variable focal length endoscope detection device according to the embodiments of the present invention, as shown in fig. 1, and the adsorption channel 1023 is located between the illumination channel and the edge of the detection tube. That is, the detection tube in the variable-focus cavity endoscope detection device provided by the embodiment of the invention is also provided with the adsorption channel 1023 for adsorbing the variable-focus cavity endoscope detection device on the tissue to be detected, and negative pressure is formed in the adsorption channel 1023 by extracting air in the adsorption channel 1023, so that the variable-focus cavity endoscope detection device is adsorbed on the tissue to be detected, wherein the adsorption channel 1023 is positioned between the illumination channel and the edge of the detection tube, namely, at a position which is positioned outside the illumination channel and is close to the side of the detection tube.

On the basis of the above embodiments, the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the invention further comprises an air extracting device, as shown in fig. 3, the air extracting device 17 mainly comprises an air extracting pump, the air extracting device is connected with the adsorption channel through an air extracting pipeline, an air extracting valve is arranged in the air extracting pipeline and is electrically connected with the air extracting device 17, the air extracting device 17 controls the air extracting flow of the air extracting pipeline by adjusting the opening and closing of the air extracting valve, so that the air extracting control of the adsorption channel is realized, the negative pressure in the adsorption channel is further adjusted, the zoom type cavity endoscope detecting device is adsorbed on tissues such as stomach intestine, mouth cavity and uterine cavity in the abdominal cavity of a human body under the action of atmospheric pressure, and motion artifacts caused by the movement of biological tissues are reduced, so that imaging is more stable and clear.

Fig. 8 is a schematic diagram of a box-type combined structure of a three-dimensional nonlinear laser scanning endoscope according to an embodiment of the present invention, as shown in fig. 8, a display 55 integrated on a box cover is integrated with a box body in which each module is installed, so that the whole equipment can be conveniently moved and a workplace can be conveniently replaced, and when the display 55 is used, the display can be externally placed on the box body, so that a worker can conveniently obtain information on the display, wherein a variable-focus endoscope detection device 1 in the three-dimensional nonlinear laser scanning endoscope is an stomatoscope detection device. After the three-dimensional nonlinear laser scanning cavity endoscope is used, a worker can carry the equipment box, and the equipment can be conveniently replaced in a workplace, especially in a hospital, a laboratory or an outdoor place.

Fig. 9 is a schematic diagram of a box-type combined structure of a three-dimensional nonlinear laser scanning endoscope according to another embodiment of the present invention, as shown in fig. 9, a display 55 integrated on a box cover is integrated with a box body provided with each module, so that the whole equipment can be conveniently moved and a workplace can be replaced, and the display 55 can be externally placed on the box body when in use, so that a worker can conveniently obtain information on the display, wherein a variable-focus endoscope detection device 1 in the three-dimensional nonlinear laser scanning endoscope is a laparoscope detection device, and a plurality of laparoscope detection devices can be simultaneously arranged. After the three-dimensional nonlinear laser scanning cavity endoscope is used, a worker can carry the equipment box, and the equipment can be conveniently replaced in a workplace, especially in a hospital, a laboratory or an outdoor place.

Based on the above embodiments, the three-dimensional nonlinear laser scanning endoscope provided by the embodiment of the invention has a plurality of variable-focus endoscope detection devices. The fluorescence collection device and the optical fiber coupling module provided by the embodiment of the invention can be simultaneously connected with a plurality of variable-focus cavity endoscope detection devices in an optical fiber communication way, namely, a plurality of detection devices are integrated in a three-dimensional nonlinear laser scanning cavity endoscope system, so that the simultaneous detection of different parts of gastrointestinal tissues is realized, and the contrast analysis is performed.

Based on the above embodiments, the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiments of the present invention further includes an adjusting optical fiber, which is used for optical fiber transmission connection between the fluorescence collection device and the optical fiber coupling module and the variable focal length cavity endoscope detection device, respectively, wherein:

the length of the adjusting optical fiber is adjustable. The optical fiber coupling module is connected with the variable-focus cavity endoscope detection device through the optical fiber with the adjustable length, so that the detection device can be flexibly moved according to different experimental scene requirements, the limit of the limited optical fiber length is avoided, the length of the adjustable optical fiber is adjustable, the application of various occasions is realized by changing the optical fibers with different lengths, and the optical fiber with different lengths can be changed at any time according to the requirements.

For the three-dimensional nonlinear laser scanning cavity endoscope provided by the above embodiments, another specific implementation manner is provided in the embodiment of the present invention, fig. 10 is a schematic diagram of a desk-top structure of the three-dimensional nonlinear laser scanning cavity endoscope provided by the embodiment of the present invention, as shown in fig. 10, the three-dimensional nonlinear laser scanning cavity endoscope includes an air extraction device 52, a first device 53, a second device 54, a display 55 and a variable focal length cavity endoscope detection device 1, where a scan acquisition controller and an industrial personal computer are integrated in the first device 53, the industrial personal computer is electrically connected with the display 55, the second device 54 is integrated with a femtosecond pulse laser, an optical fiber coupling module and a fluorescence collection device, an illumination module and an imaging module, and the optical fiber coupling module and the fluorescence collection device are all connected with an adsorption type microscope detection device 51 in an optical fiber transmission manner, where the variable focal length cavity endoscope detection device 1 is an oral cavity microscope detection device for detecting the oral cavity tissue of a human body so as to understand the information of malignancy, infiltration depth, transfer condition and presence or absence of cancer residues at the cutting edge, and the like, and the operation of the adsorption type three-dimensional nonlinear laser microscope scanning is the same as the above embodiments.

In another embodiment of the present invention, as shown in fig. 11, a table structure schematic diagram of a three-dimensional nonlinear laser scanning endoscope is provided, where the three-dimensional nonlinear laser scanning endoscope also includes an air extractor 52, a first device 53, a second device 54, a display 55, and a variable focal length endoscope detection device 1, where the first device 53 integrates a scanning acquisition controller and an industrial personal computer, the industrial personal computer is electrically connected with the display 55, the second device 54 integrates a femtosecond pulse laser, an optical fiber coupling module, a fluorescence collection device, an illumination module, and an imaging module, and the optical fiber coupling module and the fluorescence collection device are all connected with an adsorption microscope detection device 51 through optical fiber transmission, where the variable focal length endoscope detection device 1 is a laparoscope detection device, and the laparoscope detection device is embedded in a human abdomen to detect gastrointestinal tissues so as to understand information such as malignancy of a tumor, infiltration depth, a metastasis condition, and presence or absence of cancer residues at a cutting edge.

While the present invention has been described in connection with the embodiments illustrated in the drawings, it will be apparent to those skilled in the art that the present invention is not limited to the preferred embodiments of the present invention, and various modifications and variations can be made thereto by those skilled in the art on the basis of the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

The apparatus embodiments described above are merely illustrative, wherein elements illustrated as separate elements may or may not be physically separate, and elements shown as elements may or may not be physical elements, may be located in one place, or may be distributed over a plurality of network elements. Some or all of the modules may be selected according to actual needs to achieve the purpose of the solution of this embodiment. Those of ordinary skill in the art will understand and implement the present invention without undue burden.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A multiplex fluorescence collection device, comprising:

the illumination multiplexing module is used for providing illumination light signals for the remote endoscope detection device, and transmitting the first two-photon fluorescence signals and the first second harmonic signals to the dual-channel fluorescence collection module after collecting the first two-photon fluorescence signals and the first second harmonic signals based on an illumination optical fiber bundle;

the imaging multiplexing module is used for imaging a tissue region to be detected in front of an objective lens of the remote endoscope detection device, and transmitting the second two-photon fluorescence signal and the second harmonic signal to the dual-channel fluorescence collection module after collecting the second two-photon fluorescence signal and the second harmonic signal based on a bright field optical fiber bundle;

The dual-channel fluorescence collection module is used for collecting a third two-photon fluorescence signal and a third second harmonic signal based on an objective lens in a remote endoscope detection device, synchronously converging the first two-photon fluorescence signal, the first second harmonic signal, the second two-photon fluorescence signal and the second harmonic signal which are transmitted by the illumination multiplexing module and the imaging multiplexing module, converging the first two-photon fluorescence signal, the second two-photon fluorescence signal and the third two-photon fluorescence signal which are acquired by the objective lens and transmitted by the illumination multiplexing module and the imaging multiplexing module into one path, converting the converged two-photon fluorescence signal into a first electric signal, converging the first second harmonic signal, the second harmonic signal and the third second harmonic signal which are acquired by the objective lens and transmitted by the illumination multiplexing module and the imaging multiplexing module into the other path, and converting the converged second harmonic signal into a second electric signal.

2. The multiplex fluorescence collection device of claim 1, wherein the illumination multiplexing module comprises an illumination light path and a first multiplexed collection light path, wherein:

the illumination light path sequentially comprises the illumination fiber bundle, a first multiplexing dichroic mirror, a variable optical filter, an illumination lens and an illumination light source;

The first multiplexing collection optical path sequentially comprises the illumination optical fiber bundle, a first multiplexing dichroic mirror, a first multiplexing collection lens and a first transmission optical fiber.

3. The multiplex fluorescence collection device of claim 2, wherein the imaging multiplexing module comprises an imaging optical path and a second multiplexed collection optical path, wherein:

the imaging light path sequentially comprises the bright field optical fiber bundle, a second multiplexing dichroic mirror, an imaging lens and a camera;

the second multiplexing collection optical path sequentially comprises the bright field optical fiber bundle, a second multiplexing dichroic mirror, a second multiplexing collection lens and a second transmission optical fiber.

4. The multiplex fluorescence collection device of claim 3, wherein the dual channel fluorescence collection module comprises an objective collection fiber, a fiber optic universal interface, a first photomultiplier tube, a second photomultiplier tube, and a first collection optical path between the fiber optic universal interface and the first photomultiplier tube, and a second collection optical path between the fiber optic universal interface and the second photomultiplier tube, wherein:

The first collecting light path sequentially comprises a coupling lens, an infrared filter, a first dichroic mirror, a first filter and a first collecting lens, wherein the first collecting light path is used for collecting the first two-photon fluorescence signal, the second two-photon fluorescence signal and the third two-photon fluorescence signal received by the fluorescence collecting device, and the first photomultiplier is used for converting the first two-photon fluorescence signal, the second two-photon fluorescence signal and the third two-photon fluorescence signal into first electric signals;

the second collecting light path sequentially comprises the coupling lens, the infrared filter, the first dichroic mirror, the second filter and the second collecting lens, wherein the second collecting light path is used for collecting the first second harmonic signal, the second harmonic signal and the third second harmonic signal received by the fluorescent collecting device, and the second photomultiplier is used for converting the first second harmonic signal, the second harmonic signal and the third second harmonic signal into second electric signals.

5. The multiplex fluorescence collection device of claim 3, wherein the illumination multiplexing module is a plurality of modules.

6. The multiplex fluorescence collection device of claim 3, wherein the imaging multiplexing module is a plurality of modules.

7. The multiplex fluorescence collection device of claim 4, further comprising an industrial personal computer, wherein the dual-channel fluorescence collection module is in optical fiber communication with the industrial personal computer, wherein the illumination multiplexing module and the imaging multiplexing module are both in electrical connection with the industrial personal computer, and wherein the industrial personal computer is configured to obtain a first electrical signal and a second electrical signal, generate a first fluorescence image based on the first electrical signal, and generate a second fluorescence image based on the second electrical signal.

8. A three-dimensional nonlinear laser scanning cavity endoscope, comprising:

the device for detecting the cavity endoscope, the scanning acquisition controller, the femtosecond pulse laser, the optical fiber coupling module and the multi-path fluorescence collection device according to any one of claims 1 to 7, wherein the multi-path fluorescence collection device and the optical fiber coupling module are all connected with the device for detecting the cavity endoscope in an optical fiber communication manner, and the multi-path fluorescence collection device and the device for detecting the cavity endoscope are all electrically connected with the scanning acquisition controller, wherein:

the cavity endoscope detection device is used for receiving the pulse laser signal, outputting the pulse laser signal to an autofluorescent substance in a living body cell, acquiring a first two-photon fluorescence signal, a second two-photon fluorescence signal, a third two-photon fluorescence signal, a first second harmonic signal, a second harmonic signal and a third second harmonic signal which are generated after the autofluorescent substance is excited, and outputting the first two-photon fluorescence signal, the second two-photon fluorescence signal, the third two-photon fluorescence signal, the first second harmonic signal, the second harmonic signal and the third second harmonic signal to the fluorescence collection device;

9. The three-dimensional nonlinear laser scanning cavity endoscope according to claim 8, wherein the cavity endoscope detection device comprises an outer fixed housing and an inner clamping device, the inner clamping device is disposed in the outer fixed housing, a zoom motor is disposed inside the outer fixed housing for driving the inner clamping device to move up and down relative to the outer fixed housing, and an optical path structure for forming a first optical path and a second optical path is disposed in the inner clamping device, wherein:

The first optical path sequentially comprises a collimating lens, a micro-electromechanical scanning galvanometer, a lens, a dichroic mirror, a relay mirror and an objective lens, wherein the first optical path is used for conducting laser signals received by the collimating lens from the collimating lens to the objective lens;

the second optical path sequentially comprises the objective lens, the relay lens and the dichroic mirror, wherein the second optical path is used for conducting optical signals collected by the objective lens from the objective lens to the dichroic mirror.

10. The three-dimensional nonlinear laser scanning cavity endoscope of claim 9, wherein the outer fixed housing comprises a handle housing and a probe tube, the handle housing is fixedly connected with the probe tube, the zoom motor is disposed inside the handle housing, and a probe channel is disposed inside the probe tube, wherein:

the collimating lens, the micro-electromechanical scanning galvanometer, the lens and the dichroic mirror in the light path structure are all positioned in the handle shell, the relay lens and the objective lens in the light path structure are all positioned in the detection channel, and the objective lens is positioned at a channel opening of the detection channel;

a plurality of illumination channels are further arranged in the detection tube, and illumination fiber bundles for transmitting illumination light signals are arranged in the illumination channels, wherein the illumination channels are uniformly distributed by taking the axle center of the detection channel as the center;

An observation channel is further arranged in the detection tube, the observation channel is located between the detection channel and the illumination channel, an observation lens is arranged at a channel opening of the observation channel, and the observation lens is connected with a bright field optical fiber bundle in the observation channel and used for acquiring image information of a tissue region to be detected in front of the objective lens.