CN210170028U

CN210170028U - Positioning type adsorption device, microscope detection device and laser scanning microscope

Info

Publication number: CN210170028U
Application number: CN201920180481.7U
Authority: CN
Inventors: Runlong Wu; 吴润龙; Aimin Wang; 王爱民; Wenmao Jiang; 江文茂; Yanhui Hu; 胡炎辉; Heping Cheng; 程和平
Original assignee: Beijing Chaowei Landscape Biology Technology Co Ltd
Current assignee: Beijing Chaowei Landscape Biology Technology Co Ltd
Priority date: 2019-01-31
Filing date: 2019-01-31
Publication date: 2020-03-24
Anticipated expiration: 2029-01-31

Abstract

The embodiment of the utility model provides a locate mode adsorption equipment, microscope detection device and laser scanning microscope. The positioning type adsorption device comprises an outer shell, a base, a cover glass and a movement device, wherein the outer shell is detachably connected with the base through magnetic field force, and the cover glass is fixed at a sealing port of the sucker to form a built-in space and an external adsorption space; the movement device comprises a fixed support, a first driving block and a second driving block, the second driving block drives the first driving block and the fixed support to move along the Y-axis direction, and the first driving block drives the micro microscope probe to move along the X-axis direction. The embodiment of the utility model provides a locate mode adsorption equipment, microscope detecting device and laser scanning microscope realize that miniature microscope probe removes along XY axle direction in the horizontal plane through first drive block and second drive block, under unmovable adsorption equipment's the condition, look for the observation point, realize the accurate positioning of miniature microscope probe, easy operation, convenient to use.

Description

Positioning type adsorption device, microscope detection device and laser scanning microscope

Technical Field

The embodiment of the utility model provides a relate to laser scanning microscope technical field, especially relate to a locate mode adsorption equipment, microscope detection device and laser scanning microscope.

Background

With the continuous development of medicine and biology, people have made remarkable progress on the research on the cell morphology, the tissue structure or the gastrointestinal fiber state in animal life, and particularly, the related technology of obtaining the biological cell morphology of a living body by obtaining a fluorescence signal and a second harmonic signal through the excitation of pulse laser radiation in a near infrared region and the detection by a proper high-sensitivity receiver has made remarkable results.

The related detection device for acquiring the morphology of the biological cells based on the fluorescence signal, the second harmonic signal and the CARS (Coherent anti-Stokes Raman Scattering) signal plays an important role in the application of the technology. The existing imaging equipment for detecting human body cells or tissues is mainly a three-dimensional nonlinear laser scanning microscope, wherein the laser scanning microscope in the form of the existing laser scanning microscope mainly moves a microscope detection device through a mechanical arm, namely the detection device of the laser scanning microscope is arranged on the mechanical arm, the detection device is moved through the adjustment of the mechanical arm, and then different tissue structures of a human body are aligned and detected.

However, the detection device based on the mechanical arm in the three-dimensional nonlinear laser scanning microscope has a large volume, and the probe corresponds to a large skin area of a human body, so that in specific operation, specific point positions detected by the skin cannot be accurately positioned, the mechanical arm needs to be adjusted for many times, the detection device is moved for many times, the positioning effect is poor, displacement deviation is easily generated, and the imaging quality is affected.

SUMMERY OF THE UTILITY MODEL

To the technical problem who exists among the prior art, the embodiment of the utility model provides a locate mode adsorption equipment, microscope detection device and laser scanning microscope.

In a first aspect, an embodiment of the present invention provides a positioning type adsorption device capable of setting an optical fiber bundle, including:

shell body, base, coverslip and drive the telecontrol equipment that miniature microscope probe removed in the horizontal plane, be provided with the sucking disc on the base, the sucking disc embedding the sucking disc that the shell body bottom was seted up is downthehole, the shell body with the base can dismantle the connection through magnetic field force, the shell body bottom is provided with the fiber bundle hole, fiber bundle hole evenly distributed in around the sucking disc hole, wherein:

the cover glass is fixed at the sealing port of the sucker to form an internal space and an external adsorption space of the adsorption device;

the motion device set up in the built-in space, the motion device includes fixed bolster, first drive block and second drive block, first drive block with the second drive block respectively with fixed bolster fixed connection, the second drive block is fixed on the lateral wall of shell body, wherein:

the second driving block is used for driving the first driving block and the fixed support to move together in the Y-axis direction, and the first driving block is used for driving the micro microscope probe to move in the X-axis direction, wherein the micro microscope probe is aligned to the cover glass in the forward direction.

In a second aspect, an embodiment of the present invention provides an absorption microscope detecting device, including:

miniature microscope probe and the embodiment of the utility model provides a first aspect provides a locate mode adsorption equipment for setting up miniature microscope probe, miniature microscope probe is fixed on the first drive block, wherein:

miniature microscope probe includes probe housing and fixed plate, probe housing top is provided with first opening and second opening, probe housing bottom is provided with the third opening, be provided with in the third opening and be used for driving the zoom motor that objective reciprocated, the motor of zooming passes through the fixed plate can be dismantled and fix on the probe housing, wherein:

first opening with be provided with first light path in the first passageway that forms between the third opening, the second opening with be provided with the second light path in the second passageway that forms between the third opening, wherein:

the first optical path sequentially comprises a collimating lens, a micro-electromechanical scanning galvanometer, a first lens, a second lens, a dichroic mirror and the objective lens, wherein the collimating lens, the micro-electromechanical scanning galvanometer, the first lens, the second lens, the dichroic mirror and the objective lens are positioned between the first port and the third port, and the first optical path is used for conducting laser signals received by the first port from the first port to the third port;

the second optical path sequentially comprises the objective lens and the dichroic mirror between the third port and the second port, and the second optical path is used for conducting the optical signal collected by the objective lens to the second port from the third port.

In a third aspect, an embodiment of the present invention provides an adsorption type three-dimensional nonlinear laser scanning microscope, including:

multichannel fluorescence collection device, air exhaust device, scanning acquisition controller, femto second pulse laser ware, fiber coupling module and the embodiment second aspect provides an absorption formula microscope detecting device, multichannel fluorescence collection device with fiber coupling module all with absorption formula microscope detecting device optical fiber communication connects, multichannel fluorescence collection device with absorption formula microscope detecting device all with scanning acquisition controller electricity is connected, air exhaust device with absorption formula microscope detecting device electricity is connected, 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 signal output by the femtosecond pulse laser and transmitting the pulse laser signal to the first port of the micro microscope probe in the adsorption microscope detection device;

the adsorption microscope detection device is used for receiving the pulse laser signal, outputting the pulse laser signal to an autofluorescence substance in a living body cell, acquiring a fluorescence signal and a second harmonic signal generated after the autofluorescence substance is excited, and outputting the fluorescence signal and the second harmonic signal to the multi-channel fluorescence collection device;

the multi-channel fluorescence collecting device is used for receiving the fluorescence signal and the second harmonic signal and then respectively converting the fluorescence signal and the second harmonic signal into corresponding electric signals;

the scanning acquisition controller is used for controlling the micro microscope probe to scan the pulse laser signal and synchronously acquiring the electric signal;

and the air extracting device is used for extracting air from the outer adsorption space of the adsorption type microscope detection device so as to form negative pressure in the outer adsorption space.

The embodiment of the utility model provides a locate mode adsorption equipment, microscope detection device and laser scanning microscope adopts the magnetism between shell body and the base to inhale the effect and can dismantle the connection together, sucking disc and cover glass form can make the adsorption equipment adsorb the outer absorption space on the life skin and be used for placing the built-in space of miniature microscope probe, wherein, the cover glass is fixed in the sealing port of sucking disc in order to realize the sealing contact, forms above-mentioned outer absorption space through the inner space of sucking disc; the motion device is arranged in the built-in space, and the monolithic stationary is on the lateral wall of shell body, through first drive block and second drive block, realize that miniature microscope probe removes along XY axle direction in the horizontal plane, the integrated device is miniaturized, the device adsorbs the back on human skin, under the condition of unmovable adsorption equipment, look for the observation point, realize the accurate positioning of miniature microscope probe, obtain the formation of image in different regions, thereby the better imaging region that the observation wants to observe, and is easy to operate, high durability and convenient use, the accessible removes XY position simultaneously, form images a plurality of sites in proper order, obtain bigger formation of image field of vision through the image concatenation at last.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.

Fig. 1 is a schematic structural view of a positioning type adsorption device capable of arranging an optical fiber bundle according to an embodiment of the present invention;

fig. 2 is a schematic structural view of a combined positioning type adsorption device according to an embodiment of the present invention;

fig. 3 is a schematic view illustrating a positioning type adsorption apparatus provided by an embodiment of the present invention performing remote positioning by using cross laser;

FIG. 4 is a schematic view of a suction cup structure disposed on a base in a positioning type suction device according to an embodiment of the present invention;

fig. 5 is a schematic sectional structure view of a combined positioning type adsorption device according to an embodiment of the present invention;

fig. 6 is a schematic view of a cross-sectional structure of a combined positioning type adsorption device according to an embodiment of the present invention;

fig. 7 is a schematic structural diagram of a micro microscope probe according to an embodiment of the present invention;

fig. 8 is a schematic view of a first optical path structure in a probe of a micro microscope according to an embodiment of the present invention;

fig. 9 is a schematic view showing a cross-sectional structure of a combined positioning type adsorption device according to an embodiment of the present invention;

fig. 10 is a schematic cross-sectional structure view of an absorption microscope detection device provided in an embodiment of the present invention;

fig. 11 is a schematic structural view of a dual-channel fluorescence collection module in the multi-channel fluorescence collection device according to the embodiment of the present invention;

fig. 12 is a schematic view of an absorption type three-dimensional nonlinear laser scanning microscope provided by an embodiment of the present invention detecting human facial skin tissues;

fig. 13 is a schematic view of an absorption type three-dimensional nonlinear laser scanning microscope provided by the embodiment of the present invention detecting skin tissue of a chest of a human body;

fig. 14 is a schematic view of a plurality of detection devices of an absorption type three-dimensional nonlinear laser scanning microscope provided by an embodiment of the present invention detecting human skin tissues simultaneously;

fig. 15 is a schematic view of an absorption type three-dimensional nonlinear laser scanning microscope for detecting animal skin tissues according to an embodiment of the present invention;

fig. 16 is a schematic view of a box-type combination structure of an absorption type three-dimensional nonlinear laser scanning microscope according to an embodiment of the present invention;

fig. 17 is a schematic view of a box sealing structure of a box combination structure of an adsorption type three-dimensional nonlinear laser scanning microscope according to an embodiment of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, 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 obvious that the described embodiments are some, but not all, embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

The existing related detection equipment for acquiring the form of biological cells based on fluorescence signals and second harmonic signals is mainly a three-dimensional nonlinear laser scanning microscope, and the form of the laser scanning microscope at present mainly comprises a laser scanning microscope for moving a microscope detection device through a mechanical arm, namely a detection device of the laser scanning microscope is arranged on the mechanical arm, the detection device is moved through the adjustment of the mechanical arm, and then different tissue structures of a human body are aligned and detected. However, the detection device based on the mechanical arm in the three-dimensional nonlinear laser scanning microscope has a large volume, and the probe corresponds to a large skin area of a human body, so that in specific operation, specific point positions detected by the skin cannot be accurately positioned, the mechanical arm needs to be adjusted for many times, the detection device is moved for many times, the positioning effect is poor, displacement deviation is easily generated, and the imaging quality is affected.

In order to carry out accurate location formation of image to the organism tissue for the convenience, acquire its cell morphological structure information, the embodiment of the utility model provides a locate mode adsorption equipment that can set up optic fibre bundle, figure 1 does the embodiment of the utility model provides a locate mode adsorption equipment structure sketch that can set up optic fibre bundle, as shown in figure 1, this locate mode adsorption equipment includes:

outer shell body 11, base 13, coverslip 12 and drive the telecontrol equipment that miniature microscope probe 15 removed in the horizontal plane are provided with sucking disc 131 on the base 13, and the sucking disc that sets up in 11 bottoms of sucking disc 131 embedding outer shell is downthehole, and outer shell body 11 can be dismantled with base 13 through magnetic field force and be connected, the outer shell body bottom is provided with optical fiber bundle hole 10, optical fiber bundle hole 10 evenly distributed in around the sucking disc hole, wherein:

the cover glass 12 is fixed on the sealing port 1311 of the suction cup 131 to form an inner space and an outer adsorption space of the adsorption type device;

the movement device is disposed in the built-in space, and the movement device includes a fixed bracket 142, a first driving block 141 and a second driving block 143, the first driving block 141 and the second driving block 143 are respectively fixedly connected with the fixed bracket 142, and the second driving block 143 is fixed on the side wall of the outer housing 11, wherein:

the second driving block 143 is used to drive the first driving block 141 and the fixing bracket 142 to move together in the Y-axis direction, and the first driving block 141 is used to drive the micro microscope probe 15 to move in the X-axis direction, in which the micro microscope probe 15 is aligned in the normal direction with respect to the cover glass 12.

Specifically, the embodiment of the utility model provides a can set up shell body 11 and base 13 in the locate mode adsorption equipment of optic fibre bundle 15 and constitute for the material that can each other magnetism inhale, or built-in magnetic object that has each other magnetism inhale, make accessible magnetic field force can dismantle the connection together each other, and be provided with sucking disc 131 on the base 13, shell body 11 bottom is provided with the sucking disc hole that is used for imbedding sucking disc 131, be equipped with the sealing 1311 with sucking disc 131 inner space intercommunication on sucking disc 131, when cover glass 12 lid is on sealing 1311, form built-in space and can make adsorption equipment adsorb the outer space that adsorbs on the life skin in this locate mode adsorption equipment.

The moving device for driving the micro microscope probe 15 to move in the horizontal plane is arranged in the built-in space, and comprises a fixed support 142, a first driving block 141 and a second driving block 143, wherein the first driving block 141 and the second driving block 143 both comprise a sliding block and a fixed block, the first fixed block of the first driving block 141 is fixed on the fixed support 142, and the first sliding block of the first driving block 141 slides relative to the first fixed block under the driving of a first motor; the second sliding block of the second driving block 143 is fixed on the fixing bracket 142, the second fixing block of the second driving block 143 is fixed on the side wall of the outer shell 11, and the second sliding block of the second driving block 143 slides relative to the second fixing block under the driving of the second motor; the micro microscope probe 15 can be fixed on the first slider of the first driving block 141, so that the first driving block 141 can drive the micro microscope probe 15 to move along the X-axis direction, the second driving block 143 can drive the first driving block 141 and the fixing bracket 142 and the micro microscope probe 15 fixed on the first driving block 141 to move along the Y-axis direction together, so that the adsorption device can be roughly positioned on the skin tissue to be tested of the living body, and the micro microscope probe 15 can be driven to move along the X-axis and the Y-axis directions by the movement device, so that the micro microscope probe 15 can be accurately positioned in the horizontal plane, and meanwhile, the micro microscope probe 15 can be driven by the motor to move along the X-axis and the Y-axis directions, so that the imaging of different regions can be realized while the microscope adsorption device is not moved, and the imaging region desired to be observed can be better observed. Meanwhile, the displacement table can be stepped by the same distance through program control, and imaged images are spliced, so that an imaging result of N times of visual field can be obtained.

And after the fixing support 142 and the micro microscope probe 15 are fixed, the micro microscope probe 15 is aligned to the cover glass 12 in the forward direction to output an internal signal and receive an external signal through the cover glass 12, thereby realizing the zoom of the microscope and the three-dimensional imaging. And the outer shell 11, the base 13, the cover glass 12 and the moving device in the figure 1 are combined into a whole to be a positioning type adsorption device which can be provided with an optical fiber bundle 15. Fig. 2 is the embodiment of the utility model provides a locate mode adsorption equipment structural schematic diagram after the combination, as shown in fig. 2, miniature microscope probe 15 fixes on the adsorption equipment after the combination promptly for being provided with the locate mode adsorption equipment of miniature microscope probe 15, wherein miniature microscope probe 15 is located built-in space, fixes on the lateral wall of shell body 11, and miniature microscope probe 15's objective is forward to aim at the coverslip, and adsorption microscope probe's shell body 11 can be dismantled through magnetic field force with base 13 and be connected.

The positioning type adsorption device capable of setting the optical fiber bundle provided by the embodiment of the utility model adopts the magnetic attraction effect between the outer shell and the base to be detachably connected together, the sucking disc and the cover glass form an outer adsorption space which can enable the adsorption device to be adsorbed on the skin of a living body and an inner space for placing a micro microscope probe, wherein the cover glass is fixed at the sealing port of the sucking disc to realize sealing contact, and the outer adsorption space is formed through the inner space of the sucking disc; the movement device is arranged in the built-in space and integrally fixed on the side wall of the shell, the micro microscope probe moves in the horizontal plane along the XY axis direction through the first driving block and the second driving block, the whole device is miniaturized, after the device is adsorbed on the skin of a human body, the observation point is searched while the adsorption device is not moved, the precise positioning of the micro microscope probe is realized, the imaging of different areas is obtained, and therefore the better observation of the imaging area to be observed is realized, the operation is simple, and the use is convenient.

On the basis of each embodiment, the embodiment of the utility model provides a can set up locate mode adsorption equipment of optic fibre bundle, as shown in fig. 1, still include:

first word laser instrument 16 and the first word laser instrument 17 of second, first word laser instrument 16 and the first word laser instrument 17 of second all set up in shell body 11 bottom, through transmitting cover glass and outwards adsorbing the interior transmission word laser in the space, wherein:

a first in-line laser 16 for emitting in-line laser coincident with the X axis, and a second in-line laser 17 for emitting in-line laser coincident with the Y axis. Promptly for the convenient location is surveyed, the embodiment of the utility model provides a can set up the locate mode adsorption equipment's of optic fibre bundle shell body 11 bottom has still set up two word lasers, be located the X axle and the Y axle direction of sucking disc respectively, all transmit a word laser in the outside adsorption space through the transmission coverslip, a word laser and the X axle coincidence of first word laser 16 transmission, a word laser and the Y axle coincidence of a second word laser 17 transmission, thereby make the cross locating point of the cross laser that obtains coincide with the original point of XY axle and the objective optical axis center coincidence of miniature microscope probe, and the cross also coincides with XY axle. Fig. 3 is the embodiment of the utility model provides a locate mode adsorption equipment carries out remote location sketch map through cross laser, as shown in fig. 3, when locate mode adsorption equipment 1 that has miniature microscope probe is far away from the skin tissue of the life body that awaits measuring, the accessible transmission advances the cross laser in the outer adsorption space and carries out remote location, then passes through the telecontrol equipment again, carries out the accurate positioning of XY axle direction to more convenient, the accurate imaging information who acquires different regions.

miniature camera 18 and illumination fiber optic bundle 101, miniature camera 18 and illumination fiber optic bundle 101 all set up in the shell body bottom, wherein:

the miniature camera 18 is a miniature CCD or a miniature CMOS;

the lens of the miniature camera 18 faces the outer adsorption space, and the angle of the lens of the miniature camera 18 is adjustable;

the illumination fiber bundle 101 is disposed in the fiber bundle hole 10, and is used for emitting an optical signal for providing illumination into the outer absorption space. In order to make things convenient for the accurate positioning more promptly, the embodiment of the utility model provides a can set up the locate mode adsorption equipment's of optic fibre bundle 15 shell body 11 bottom has still set up miniature camera 18 and illumination optic fibre bundle 101, use distortionless wide-angle lens before the miniature camera 18, the camera lens is towards outside absorption space, see through the coverslip and can obtain great field of vision, when adsorption equipment adsorbs on skin, can observe the relative position of objective and skin of miniature microscope probe 15 through miniature camera 18, rethread telecontrol equipment moves miniature microscope probe 15 to the position of required observation along XY axle direction. The lens angle of the miniature camera 18 can be adjusted up and down according to the requirement; the illuminating fiber bundle 101 can be used for absorbing light signals emitted from the space to the outside, and providing illumination for the miniature camera 18 when the absorbing device is absorbed on the skin, wherein the wavelength of the illuminating fiber bundle for providing illumination is separated from the wavelength of the fluorescence imaging, for example, the wavelength of the second harmonic signal for imaging is 390nm and the wavelength of the two-photon autofluorescence signal for imaging is 450-600nm, and the wavelength of the LED lamp can be selected from 370nm, 635nm or infrared 850nm, 940 nm.

On the basis of the above embodiments, the suction cup in the positioning type suction device capable of setting the optical fiber bundle provided by the embodiment of the present invention further includes a suction port and an air exhaust port, fig. 4 is a schematic view of a suction cup structure arranged on a base in the positioning type suction device provided by the embodiment of the present invention, as shown in fig. 4, the suction port 1312 is communicated with the sealing port 1311 for being sucked on a living body to be measured through an external suction space;

the pumping port 1313 communicates with the outer adsorption space to pump the gas in the outer adsorption space. That is, the suction cup 131 disposed on the base 13 in the absorption microscope detecting device provided by the embodiment of the present invention further includes an absorption port 1312 and an extraction port 1313 in addition to the sealing port 1311, and the sealing port 1311 is communicated with the absorption port 1312 to form an internal space of the suction cup 131, when the cover glass is hermetically fixed on the sealing port 1311, the internal space of the suction cup 131 forms an external absorption space, and the absorption port 1312 is absorbed on the living body to be measured through the external absorption space; the air exhaust port 1313 communicates with the outer adsorption space, and the air in the outer adsorption space can be exhausted through the air exhaust port 1313, thereby forming a negative pressure in the outer adsorption space. Under the action of external pressure, the adsorption type microscope detection device is adsorbed on tissues such as skin of a to-be-detected living body.

Wherein, fig. 5 is the schematic view of the sectional structure of the positioning type adsorption device after combination, as shown in fig. 5, the suction cup is embedded into the suction cup hole of the outer shell 11, the cover glass 12 is fixed on the suction cup, the micro microscope probe 15 is fixed on the side wall of the outer shell 11 through the moving device 14, and the objective of the micro microscope probe 15 is aligned to the cover glass 12 in the forward direction. The micro microscope probe 15 is located in the built-in space 1314, the absorption type microscope detection device is absorbed on tissues such as skin of a to-be-detected living body through negative pressure formed in the outer absorption space 1315, an air suction opening 1313 of the suction cup is communicated with a through hole 1111 in the outer shell and used for placing an air suction pipeline, and the first linear laser 16 and the micro camera 18 are fixed at the bottom of the outer shell 11.

Wherein, fig. 6 is the schematic view of the sectional structure of the positioning type adsorption device after combination, as shown in fig. 6, the micro microscope probe 15 is located in the built-in space, and the objective lens of the micro microscope probe 15 is directed to the cover glass, and the second linear laser 17 and the illumination fiber bundle 101 are fixed at the bottom of the outer shell 11.

On the basis of the above embodiments, the embodiment of the present invention provides a fixing bracket in a positioning type absorption device capable of setting an optical fiber bundle, including an X-axis bracket and a Y-axis bracket, wherein the X-axis bracket and the Y-axis bracket are mutually perpendicular and fixedly connected, and wherein:

the X-axis support is detachably and fixedly connected with the first driving block, and the Y-axis support is detachably and fixedly connected with the second driving block. That is to say the embodiment of the utility model provides a can set up fixed bolster in the location formula adsorption equipment of optic fibre bundle is the L type, and it includes X-axis support and Y-axis support, X-axis support and Y-axis support mutually perpendicular fixed connection, and fixed connection can be dismantled with the first mount of first drive block to X-axis support, and fixed connection can be dismantled with the second slider of second drive block to realize that miniature microscope probe can follow XY axle direction and remove under telecontrol equipment's drive.

The embodiment of the utility model provides a still provide an absorption formula microscope detection device, this absorption formula microscope detection device includes:

the micro microscope probe and the positioning type adsorption device which is provided by the above embodiments and can be provided with the optical fiber bundle, the micro microscope probe is fixed on the first driving block, wherein:

miniature microscope probe is including probe housing and fixed plate, and probe housing top is provided with first opening and second opening, and probe housing bottom is provided with the third opening, is provided with in the third opening to be used for driving the zoom motor that objective reciprocated, and the motor of zooming can be dismantled through the fixed plate and fix on probe housing, wherein:

be provided with first light path in the first passageway that forms between first opening and the third opening, be provided with the second light path in the second passageway that forms between second opening and the third opening, wherein:

the first light path sequentially comprises a collimating lens, a micro-electromechanical scanning galvanometer, a first lens, a second lens, a dichroic mirror and an objective lens, wherein the collimating lens, the micro-electromechanical scanning galvanometer, the first lens, the second lens, the dichroic mirror and the objective lens are positioned between the first port and the third port;

the second optical path comprises an objective lens and a dichroic mirror which are arranged between the third port and the second port in sequence, wherein the second optical path is used for conducting optical signals collected by the objective lens to the second port from the third port.

Specifically, fig. 7 is a schematic structural diagram of a micro microscope probe according to an embodiment of the present invention, as shown in fig. 7, three openings are disposed on a probe housing 151 of the micro microscope probe, two channels are formed between the three openings, two optical paths are disposed in the two channels, which are respectively a first optical path and a second optical path, a section of overlapped portion is disposed between the two channels formed between the three openings, and a dichroic mirror 1544 and an objective 1545 shared by the two optical paths are disposed in the overlapped portion of the channels, wherein the first optical path is an emission optical path, and sequentially includes a collimating lens 157 located between the first opening 154 and the third opening 156, a micro electro mechanical scanning galvanometer 1541, a first lens 1542, a second lens 1543, a dichroic mirror 1544, and an objective 1545, that is, a laser signal enters from the first opening 154, is conducted through the first optical path, and exits from the third opening 156, parallel light incident to the first through opening is emitted in a parallel light mode after passing through the two lenses; the second optical path is a collecting optical path and is used for outputting the collected optical signals to a collecting device connected with the probe of the micro microscope from the second port 155; the objective lens shared by the first optical path and the second optical path is installed in a high-precision zoom motor 153, and can move up and down under the driving of the zoom motor, wherein the zoom motor 153 is detachably fixed on the probe shell 151 through a fixing plate 152; and the laser signal that the first light path transmits is under the reflection of dichroic mirror 1544, can be incident into objective 1545 perpendicularly forward, the dichroic mirror reflects the laser signal of the first light path, transmit the optical signal that the second light path gathers, and the pitch angle of this dichroic mirror 1544 is adjustable, objective 1545 is the infinity objective, wherein the step precision of the above-mentioned zooming motor is less than 1 micron.

Wherein, fig. 8 is the first light path structure sketch map in the miniature microscope probe that the embodiment of the utility model provides, as shown in fig. 8, the utility model provides an in the miniature microscope probe second focus 73 of first lens 71 and the coincidence of the first focus of second lens 72, micro-electromechanical scanning galvanometer lie in first focus 75 of first lens 71, the back pupil face 74 of objective lie in the second focus of second lens 72 to form the conjugation between the light beam of incidenting to micro-electromechanical scanning galvanometer and the light beam of output to objective back pupil face 74. Namely, the embodiment of the utility model provides an among the first light path structure in the miniature microscope probe, a micro-electromechanical scanning galvanometer for scanning incident laser signal installs in first focus department 75 of first lens 71, the coincidence of the first focus of second focus 73 of first lens 71 and second lens 72, through adjusting the zoom motor, make the back pupil face 74 of objective be located the second focus department of second lens 72, thereby realize, form the conjugation between the light beam of incidenting to micro-electromechanical scanning galvanometer and the light beam of output to on the back pupil face 74 of objective, thereby make D₂/D₁＝F₂/F₁Wherein D is₂Is the spot diameter, D, of the beam incident on the objective lens back pupil surface 74₁Is the spot diameter, F, of the light beam incident on the first focal plane of the first lens 71₂Is the focal length, F, of the second lens 72₁Is the focal length of the first lens 71; tan theta₂/tanθ₁＝F₁/F₂，θ₁Is the scan angle, θ, of the incident beam at the first focal plane of the first lens 71₂For the scanning angle of the incident beam at the objective lens back pupil surface 74, the incident laser beam is expanded by adjusting the first lens 71 and the second lens 72, so that the beam entering the objective lens fills the whole objective lens back pupil surface 74, to realize high-resolution imaging.

The embodiment of the utility model provides an absorption formula microscope detection device adopts the miniature microscope probe that installs objective additional in the locate mode absorption device and can adjust from top to bottom, move in the miniature microscope probe horizontal plane of movement device drive and carry out different imaging region accurate positioning, reciprocate through zooming motor drive miniature microscope probe, carry out the accurate detection of the different degree of depth to the life body cell form that awaits measuring, under the inter combination, thereby realize surveying the three-dimensional accurate positioning of the different regional different degree of depth of life body tissue, acquire the high resolution imaging of the cell structure of the different regional different degree of depth, and the method is simple and convenient to operate.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides an absorption formula microscope detection device still includes:

the liquid lens is positioned between the collimating lens and the micro-electromechanical scanning galvanometer to form a new first light path, and the new first light path sequentially comprises the collimating lens, the liquid lens, the micro-electromechanical scanning galvanometer, the first lens, the second lens, the dichroic mirror and the objective lens, wherein the collimating lens is positioned between the first port and the third port. That is to say, the embodiment of the present invention provides an multiplicable liquid lens 158 of first light path in the miniature microscope probe in the absorption type microscope detection device, fig. 9 is the embodiment of the present invention provides a schematic diagram of the section structure of the positioning type absorption device after combination, as shown in fig. 9, through increasing the liquid lens 158 on the miniature microscope probe 15 structure shown in fig. 9, the function of zooming is realized. The liquid lens 158 is located between the collimating lens 157 and the micro-electromechanical scanning galvanometer 1541, and in the primary optical path structure shown in fig. 9, the laser signal is parallel light after passing through the collimating lens 157. After the liquid lens 158 is added, a voltage or a current is applied to the liquid lens 158 to generate a corresponding curvature on the surface of the liquid lens 158, thereby generating different optical powers for the parallel light. The specific light path is as follows: laser signals are emitted from the optical fibers, parallelly enter the liquid lens 158 after passing through the collimating lens, corresponding focal power is generated from the liquid lens 158 according to loaded voltage or current signals, and emitted convergent or divergent light is converged on a sample after passing through the micro-electro-mechanical scanning galvanometer 1541, the first lens, the second lens, the dichroic mirror and the objective lens. The focal power change introduced by the liquid lens 158 can make the focal point of the laser signal emitted from the objective lens port move back and forth in the depth direction, and the liquid lens 158 has a very fast response speed, and the scanning frequency is in the order of KHz, so that fast depth direction scanning imaging can be realized. The liquid lens 158 is equivalent to a parallel flat glass when no voltage or current signal is applied, and has no optical power for the laser signal and does not cause any shift of the focus behind the objective lens. In specific use, the liquid lens 158 is complementary to a zoom motor, the position of an objective lens is adjusted through the zoom motor, after the objective lens is roughly adjusted to a corresponding depth position, the system is switched to a liquid lens 158 zoom scanning mode, and a sample is rapidly imaged in three dimensions, wherein when the adsorption type microscope detection device is not provided with the zoom motor, the zoom adjustment can be performed only through the liquid lens.

On the basis of above-mentioned each embodiment, among the absorption formula microscope detection device that the embodiment of the utility model provides an objective outside of miniature microscope probe is provided with the conduction fiber bundle all around, the conduction fiber bundle be used for the light signal before the objective extremely the dichroic mirror. That is to say in the absorption formula microscope detection device that the embodiment provided still be provided with conduction fiber bundle around the objective outside of miniature microscope probe, figure 10 is the utility model provides an absorption formula microscope detection device section structure sketch map, as shown in figure 10, the objective 1545 outside of miniature microscope probe still is provided with a large amount of conduction fiber bundle 102 all around, and the light signal before this conduction fiber bundle 102 conduction objective 1545 includes fluorescence signal and second harmonic signal to dichroic mirror to realize more light signal collection.

On the basis of the above embodiments, the outer shell in the positioning type absorption device capable of setting the optical fiber bundle provided by the embodiments of the present invention includes a first shell and a second shell, as shown in fig. 1, wherein:

an accommodating space is arranged in the first shell 111, the movement device is arranged in the accommodating space, and the first shell 111 and the second shell 112 are detachably and fixedly connected. That is, the embodiment of the present invention provides an outer casing in a positioning type absorption device capable of setting an optical fiber bundle, which comprises two parts, namely a first casing 111 and a second casing 112, wherein the first casing 111 has a containing space for setting a movement device, that is, the sucker hole in the above embodiment is also arranged at the bottom of the first casing 111, the sucker is embedded into the sucker hole of the first casing 111, and then the cover glass and the second casing 112 are combined to form an outer absorption space and a built-in space in the above embodiment, wherein the first casing 111 and the second casing 112 can be detached by screws to be fixedly connected, so as to facilitate the assembly, disassembly and replacement of components of the whole device.

The embodiment of the utility model provides a still provide an absorption formula three-dimensional nonlinear laser scanning microscope, this laser scanning microscope includes:

multichannel fluorescence collection device, air exhaust device, scanning acquisition controller, femto second pulse laser ware, the absorption formula microscope detection device that the above-mentioned embodiment provided, multichannel fluorescence collection device and optical fiber coupling module all with absorption formula microscope detection device optical fiber communication connection, multichannel fluorescence collection device and absorption formula microscope detection device all are connected with scanning acquisition controller electricity, air exhaust device is connected with absorption formula microscope detection device electricity, wherein:

the optical fiber coupling module is used for coupling the pulse laser signal output by the femtosecond pulse laser and transmitting the pulse laser signal to a first port of a micro microscope probe in the adsorption type microscope detection device;

the absorption type microscope detection device is used for receiving the pulse laser signal, outputting the pulse laser signal to the autofluorescence substance in the cells of the living body, acquiring a fluorescence signal and a second harmonic signal generated after the autofluorescence substance is excited through an objective lens, and outputting the fluorescence signal and the second harmonic signal to the multi-channel fluorescence collection device;

the scanning acquisition controller is used for controlling the micro microscope probe to scan the pulse laser signal and synchronously acquiring an electric signal;

and the air extractor is used for extracting air from the outer adsorption space of the adsorption type microscope detection device so as to form negative pressure in the outer adsorption space.

Specifically, the embodiment of the present invention provides an absorption type three-dimensional nonlinear laser scanning microscope, which comprises a multi-channel fluorescence collecting device, an air extractor, a scanning collecting controller, a femtosecond pulse laser, a fiber coupling module and an absorption type microscope detecting device, thereby forming a three-dimensional nonlinear laser scanning microscope capable of being absorbed on human skin or penetrating into intestines and stomach of human body for detection, wherein the femtosecond pulse laser can emit pulse laser signals for exciting autofluorescence in human skin cells, generating multiphoton fluorescence signals and second harmonic signals, including using 920nm femtosecond pulse laser to excite FAD and collagen in cells, exciting 500-600nm fluorescence signals and 460nm second harmonic signals, and exciting autofluorescence such as FAD or NADH in cells through 780nm femtosecond pulse laser, to generate corresponding fluorescent and second harmonic signals;

the multi-path fluorescence collecting device integrates two paths of signal collecting light paths, namely a fluorescence signal collecting light path and a second harmonic signal collecting light path, so as to realize the respective collection of fluorescence signals and second harmonic signals; the scanning acquisition controller controls a scanning galvanometer in the probe of the micro microscope to scan the pulse laser signal and excite the self-luminous fluorescent substance to generate a fluorescent signal and a second harmonic signal, and acquires a first electric signal and a second electric signal which are obtained by converting the fluorescent signal and the second harmonic signal by the multi-channel fluorescent collection device; the air extracting device mainly comprises an air extracting pump connected with an air extracting pipeline, the air extracting pipeline is connected with the air extracting opening in the embodiment, an air extracting valve is arranged in the air extracting pipeline, the air extracting valve is electrically connected with the air extracting device, the air extracting device controls the air extracting flow of the air extracting pipeline by adjusting the opening and closing of the air extracting valve, thereby realizing the air extracting control of the external adsorption space, further adjust the negative pressure in the external adsorption space to ensure that the adsorption device is adsorbed on the tissues of the skin, the intestines and stomach and the like of the living body under the action of the atmospheric pressure, and the absorption type three-dimensional nonlinear laser scanning microscope can comprise a two-photon scanning microscope, a multi-photon scanning microscope and the like according to the classification, wherein, when the femtosecond pulse laser can be replaced by a common continuous laser, an aperture diaphragm is added, and the adsorption type three-dimensional nonlinear laser scanning microscope can be adjusted to be a confocal microscope. The resolution of the adsorption type three-dimensional nonlinear laser scanning microscope can be set to 800nm, the imaging field can be 200 micrometers by 200 micrometers, and the imaging speed can be 26 frames (256 pixels by 256) or frames (512 pixels by 512 pixels).

The embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope adopts multichannel fluorescence collection device, air exhaust device, scanning acquisition controller, femto second pulse laser, fiber coupling module and absorption formula microscope detection device, thereby form can adsorb the three-dimensional nonlinear laser scanning microscope that human skin or deepen human intestines and stomach go on surveying, adjust the focus through the distance of adjusting micro microscope probe and cover glass, realize laser scanning microscope's three-dimensional scanning, arouse intracellular fluorescence material spontaneous acquisition multiphoton fluorescence signal and second harmonic signal through femto second pulse laser, realize laser scanning microscope nonlinearity, collect fluorescence signal and second harmonic signal through multichannel fluorescence collection device, and convert corresponding electric signal, and then obtain corresponding fluorescence image etc. that reflects cell tissue structure through this electric signal, wherein, the adoption of absorption formula microscope detection device can avoid the organism activity to produce the vibration influence to miniature microscope probe, thereby avoid the vibration to influence the formation of image quality, and above-mentioned laser scanning microscope can realize multiple imaging mode and include XY formation of image, XZ formation of image and 3D formation of image, wherein XY formation of image is for carrying out the horizontal scanning formation of image on certain floor of certain degree of depth of cell structure, XZ formation of image is from the XZ section formation of image of certain degree of depth down on the top layer, 3D formation of image is from certain degree of depth down on the top layer, all carry out XY formation of image on every degree of depth, the reconsitution becomes 3D image, equipment operation is simple, high durability and convenient use.

On the basis of each embodiment, the embodiment of the utility model provides a multichannel fluorescence collection device includes:

two-channel fluorescence collection module and illumination multiplex module, illumination multiplex module with two-channel fluorescence collection module optical fiber communication connection, wherein:

the illumination multiplexing module is used for providing an illumination light signal of the adsorption type microscope detection device and collecting a two-photon fluorescence signal and a second harmonic signal based on an illumination optical fiber beam;

the double-channel fluorescence collection module is used for collecting two-photon fluorescence signals and second harmonic signals by the probe of the micro microscope and synchronously converging the two-photon fluorescence signals and the second harmonic signals collected by the illumination multiplexing module to be converted into corresponding electric signals. That is, the multi-channel fluorescence collecting device provided by the embodiment of the present invention mainly comprises a dual-channel fluorescence collecting module and an illumination multiplexing module, the illumination multiplexing module is connected with the dual-channel fluorescence collecting module through optical fiber communication, the illumination multiplexing module can collect partial fluorescence signals including two-photon fluorescence signals and second harmonic signals, and simultaneously, the illumination multiplexing module also has a function of providing illumination light signals for the far-end endoscope detecting device, the illumination multiplexing module transmits the collected two-photon fluorescence signals and second harmonic signals to the dual-channel fluorescence collecting module through optical fibers, the dual-channel fluorescence collecting module collects the two-photon fluorescence signals and second harmonic signals through an objective lens in the far-end endoscope detecting device and synchronously collects the two-photon fluorescence signals and second harmonic signals collected by the illumination multiplexing module, and converts the two-photon fluorescence signals and the second harmonic signals into corresponding electrical signals, so as to acquire corresponding cell structure images through electric signals.

On the basis of each embodiment, fig. 11 is the utility model provides a binary channels fluorescence collection module structure sketch map among multichannel fluorescence collection device, as shown in fig. 11, among the three-dimensional nonlinear laser scanning microscope of absorption formula that the embodiment of the utility model provides an among the binary channels fluorescence collection module include optic fibre general interface 881, first photomultiplier 882, second photomultiplier 883 and be located optic fibre general interface 881 and the first photomultiplier 882 between first collection light path, be located the second between optic fibre general interface 881 and the second photomultiplier 883 and collect the light path, wherein:

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 the fluorescent signal received by the fluorescent collecting device, and the first photomultiplier 882 is used for converting the fluorescent signal into a first electrical signal;

the second collecting light path sequentially comprises 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, wherein the second collecting light path is used for collecting a second harmonic signal received by the fluorescence collecting device, and the second photomultiplier 883 is used for converting the second harmonic signal into a second electrical signal;

the illumination multiplexing module comprises an illumination light path and a first multiplexing collection light path, wherein:

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

the first multiplexing collecting optical path sequentially comprises a lighting optical fiber bundle, a first multiplexing dichroic mirror, a first multiplexing collecting lens and a first transmission optical fiber. That is, in the multi-channel fluorescence collecting device provided by the embodiment of the present invention, the two-channel fluorescence collecting module integrates two-channel signal collecting optical paths, which are respectively a fluorescence signal collecting optical path and a second harmonic signal collecting optical path, to realize the respective collection of fluorescence signals and second harmonic signals, wherein the processed fluorescence signals and second harmonic signals are from the fluorescence signals and second harmonic signals collected by the objective lens in the remote endoscope detecting device, and the fluorescence signals and second harmonic signals converged into the two-channel fluorescence collecting module by the illumination multiplexing module and the imaging multiplexing module, wherein, as shown in fig. 10, the first dichroic mirror 83 in the first collecting optical path is a transmission fluorescence signal, the dichroic mirror reflecting the second harmonic, the second dichroic mirror 86 and the first dichroic mirror 83 are the same dichroic mirror for reflecting the second harmonic, the first optical filter 84 is used for transmitting the fluorescence signals, the second optical filter 87 is used for transmitting corresponding second harmonic signals to filter the remaining interference signals, for example, when 780nm femtosecond fiber laser is used for exciting autofluorescence substances in abdominal cavities or oral cells of human bodies, 390nm second harmonic signals and 450-plus-600 nm two-photon autofluorescence signals can be obtained, wavelengths above 420nm pass through, a dichroic mirror reflected by wavelengths below 420, namely a first dichroic mirror 83, can separate two paths of fluorescence, and 390 +/-20 nm first optical filter 84 and 450-plus-600 nm second optical filter 87 are respectively used for obtaining clean second harmonic signals and fluorescence signals;

the embodiment of the utility model provides an in multichannel fluorescence collection device illumination multiplex module has illumination and fluorescence collection function, and the light path that corresponds is illumination light path and first multiplexing collection light path, wherein, illumination light path includes illumination fiber bundle, first multiplexing dichroic mirror, variable optical filter, illumination lens and light source, and light source launches illumination light signal to illumination lens, assembles to variable optical filter, after filtering, transmits first multiplexing dichroic mirror, transmits to distal end endoscope detection device through illumination fiber bundle, provides the illumination for endoscope detection device, simultaneously, first multiplexing collection light path includes illumination fiber bundle, first multiplexing dichroic mirror, first multiplexing collection lens and first transmission optic fibre in proper order, illumination fiber bundle gathers the two-photon fluorescence signal before distal end endoscope detection device and the second harmonic signal is transmitted to first multiplexing dichroic mirror, the illumination light source is reflected to a first multiplexing collecting lens through a first multiplexing dichroic mirror, converged to a first transmission optical fiber, and transmitted to a dual-channel fluorescence collecting module through the first transmission optical fiber, wherein the illumination light source can switch different optical filters through an electric variable optical filter rotating wheel to obtain illumination light signals with different wavelengths, the basic principle is that two-photon fluorescence imaging is not interfered, for example, when autofluorescence and second harmonic are obtained, the optical filters can be switched to red or infrared to obtain illumination light signals with 370nm, 635nm, infrared 850nm and 940nm, and the illumination light signals enter an illumination optical fiber bundle through lens coupling.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides an adsorption type three-dimensional nonlinear laser scanning microscope still includes the industrial computer, and the industrial computer is connected with scanning acquisition controller electricity, wherein:

the industrial personal computer is used for acquiring a first electric signal and a second electric signal acquired by the scanning acquisition controller, generating a first fluorescence image based on the first electric signal and generating a second fluorescence image based on the second electric signal. Namely the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes the industrial computer of being connected with scanning acquisition controller electricity, and this industrial computer generates first fluorescence image and generates second fluorescence image based on the second signal of telecommunication based on first signal of telecommunication, can be used for showing cell structure and fiber structure information respectively, wherein installs control software on the industrial computer, through control software, sends control instruction to the scanner to control scanning acquisition controller, acquire above-mentioned first signal of telecommunication and second signal of telecommunication.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes the display, and the display is connected with the industrial computer electricity for show first fluorescence image and second fluorescence image. Namely the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes the display that is used for showing first fluorescence image and second fluorescence image, through the display, the staff can directly acquire the relevant information of first fluorescence image and second fluorescence image.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides an among the absorption formula three-dimensional nonlinear laser scanning microscope absorption formula microscope detection device be a plurality ofly. Namely the embodiment of the utility model provides a multichannel fluorescence collection device and fiber coupling module can be simultaneously with a plurality of absorption formula microscope detection device optical fiber communication connection, a plurality of detection device of integration in an absorption formula three-dimensional nonlinear laser scanning microscope system promptly to the realization is surveyed the different tissue positions of life body simultaneously, thereby carries out contrastive analysis.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes adjusting optic fibre for multichannel fluorescence collection device and fiber coupling module respectively with absorption formula microscope detection device between the optical fiber transmission be connected, wherein:

the length of the adjusting optical fiber can be adjusted. Namely the embodiment of the utility model provides a multichannel fluorescence collection device and optical fiber coupling module among absorption formula three-dimensional nonlinear laser scanning microscope carry out optical fiber transmission through adjustable length's regulation optic fibre and absorption formula microscope detection device respectively and are connected, in order to realize according to different experiment scene needs, carry out nimble detection device that removes, avoid limited optical fiber length's restriction, wherein, the length adjustable of adjusting optic fibre, for the optic fibre through changing different length, realize the application of various occasions, can carry out the optic fibre change of different length as required at any time.

To illustrate the application scenario of the adsorption type three-dimensional nonlinear laser scanning microscope provided by the embodiment of the present invention more clearly, a legend is further used for illustration, fig. 12 is a schematic diagram of the adsorption type three-dimensional nonlinear laser scanning microscope provided by the embodiment of the present invention for detecting the skin tissue of the face of a human body, as shown in fig. 12, the adsorption type microscope detecting device 51 is adsorbed on the face of a human body through the air-extracting function of the air-extracting device 52, wherein, 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 and a multi-path fluorescence collecting device, the optical fiber coupling module and the multi-path fluorescence collecting device are all in optical fiber transmission connection with the adsorption type microscope detecting device 51, wherein, the working principle of the adsorption type three-dimensional nonlinear laser scanning microscope is the same, and will not be described in detail herein.

Wherein, fig. 13 is the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope surveys human chest skin tissue schematic diagram, as shown in 13, through air exhaust device 52's air exhaust function, adsorb absorption formula microscope detection device 51 at human chest, wherein, scan acquisition controller and industrial computer have been integrated in first device 53, the industrial computer is connected with display 55 electricity, second device 54 has integrated femto second pulse laser, fiber coupling module and multichannel fluorescence collection device all are connected with absorption formula microscope detection device 51 optical fiber transmission, wherein, this absorption formula three-dimensional nonlinear laser scanning microscope theory of operation is the same with above-mentioned each embodiment, here is no longer repeated.

Wherein, fig. 14 is a schematic view of a plurality of detection devices of an absorption type three-dimensional nonlinear laser scanning microscope provided by an embodiment of the present invention for simultaneously detecting skin tissue of a human body, as shown in fig. 14, a plurality of absorption type microscope detection devices 51 are respectively and simultaneously absorbed on the face, the chest and the legs of the human body by the air-extracting function of an air-extracting device 52, wherein a scanning acquisition controller and an industrial control computer are integrated in a first device 53, the industrial control computer is electrically connected with a display 55, a femtosecond pulse laser, an optical fiber coupling module and a multi-path fluorescence collection device are integrated in a second device 54, the optical fiber coupling module and the multi-path fluorescence collection device are both in optical fiber transmission connection with the absorption type microscope detection devices 51, thereby, the purpose of simultaneously detecting skin tissue structures of different parts of the human body under the effect of the plurality of absorption type microscope detection devices, and because the absorption type microscope detection device is connected with the second device by adopting optical fiber transmission, the length of the optical fiber can be adjusted, so that the human body to be detected can move freely. The working principle of the absorption type three-dimensional nonlinear laser scanning microscope is the same as that of the above embodiments, and the details are not repeated here. Fig. 15 is the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope surveys animal skin tissue sketch map, as shown in fig. 15, equally can be through air exhaust device 52's air exhaust function, adsorb absorption formula microscope detection device 51 on the skin tissue of life entity, wherein, scan acquisition controller and industrial computer have been integrated in first device 53, the industrial computer is connected with display 55 electricity, second device 54 has integrateed femto second pulse laser, fiber coupling module and multichannel fluorescence collection device all with absorption formula microscope detection device 51 optical fiber transmission connection, its theory of operation is the same with above-mentioned each embodiment.

To the three-dimensional nonlinear laser scanning microscope of absorption formula that each above-mentioned embodiment provided, the utility model discloses the embodiment still provides another embodiment, fig. 16 is the utility model discloses the embodiment provides box integrated configuration schematic diagram of three-dimensional nonlinear laser scanning microscope of absorption formula, as shown in fig. 16, this three-dimensional nonlinear laser scanning microscope's of absorption formula scanning acquisition controller 531, industrial computer 532, air exhaust device 52, illumination multiplex module 57, binary channels fluorescence collection module 56 and femto second pulse laser and fiber coupling integrated module 540 together, integrate together in portable suitcase, the incasement has the industrial computer of taking the display screen, and integrated display 55 on the case lid of suitcase; the absorption type microscope detection device 51 is absorbed on the skin tissue of a human body to be detected, is in optical fiber coupling with the multi-path fluorescence collection device 56 in the box body, is in optical fiber communication connection with the air pump through an air suction pipeline, and is electrically connected with the scanning acquisition controller 531 and the industrial personal computer 532 through a power plug. Wherein figure 17 is the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope's box integrated configuration's joint sealing structure sketch map, as shown in figure 17, integrated display 55 is in the same place with the box integration of installing each module on the case lid, makes things convenient for whole equipment to remove to and change workplace, and this display 55 when using, can place outward on the box, in order to make things convenient for the staff to acquire the information on the display. After the adsorption type three-dimensional nonlinear laser scanning microscope is used, a worker can carry the equipment box by hand, so that the workplace can be conveniently replaced, and particularly, the equipment can be more conveniently used in hospitals, laboratories or outdoor places.

It should be further noted that, in the adsorption type three-dimensional nonlinear laser scanning microscope provided in the above embodiments, after changing the wavelength of the femtosecond pulse laser and adjusting the filtering range of each filter, the CARS signal can be collected on the tissue with partial fluorescence and non-SHG (second harmonic Generation) signal activity, so as to adjust the adsorption type micro CARS microscope, and specific adjustment parameters can be set according to specific requirements.

Although the present invention has been described with reference to the accompanying drawings, it is not intended to limit the scope of the invention, and it should be understood by those skilled in the art that the above is not required to be limited to the preferred embodiments of the present invention, but rather, the present invention is not limited to the above embodiments. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

The above-described embodiments of the apparatus are merely illustrative, and the units described as separate parts may or may not be physically separate, and the parts displayed as units may or may not be physical units, may be located in one place, or may be distributed on a plurality of network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the solution of the present embodiment. One of ordinary skill in the art can understand and implement it without inventive effort.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention in its corresponding aspects.

Claims

1. A positioning type adsorption device capable of arranging an optical fiber bundle is characterized by comprising:

the cover glass is fixed at the sealing port of the sucker to form an internal space and an external adsorption space of the positioning type adsorption device;

the second driving block is used for driving the first driving block and the fixed support to move together along the Y-axis direction, and the first driving block is used for driving the micro microscope probe to move along the X-axis direction, wherein the micro microscope probe is aligned to the cover glass in the forward direction.

2. The apparatus of claim 1, further comprising:

first word laser instrument and first word laser instrument of second, first word laser instrument with first word laser instrument of second all set up in the shell body bottom, through the transmission the cover glass to transmit a word laser in the outer absorption space, wherein:

the first in-line laser is used for emitting in-line laser coincident with the X axis; and the second linear laser is used for emitting linear laser superposed with the Y axis.

3. The apparatus according to claim 1 or 2, further comprising:

miniature camera and illumination fiber bundle, miniature camera with the illumination fiber bundle all set up in the shell body bottom, wherein:

the miniature camera is a miniature CCD or a miniature CMOS;

the lens of the miniature camera faces the outer adsorption space, and the angle of the lens of the miniature camera can be adjusted;

the illumination optical fiber bundle is arranged in the optical fiber bundle hole and used for transmitting an optical signal for providing illumination into the outer adsorption space.

4. The positioning type adsorption device capable of arranging the optical fiber bundle according to claim 1 or 2, wherein the suction cup further comprises an adsorption port and an air suction port, the adsorption port is communicated with the sealing port and is used for adsorbing a living body to be tested through the outer adsorption space;

the air pumping port is communicated with the outer adsorption space and used for pumping the air in the outer adsorption space.

5. An absorption microscope probe, comprising:

the micro microscope probe and the positioning type adsorption device capable of arranging the optical fiber bundle of any one of claims 1 to 4, wherein the micro microscope probe is fixed on the first driving block, and the micro microscope probe is fixed on the first driving block, wherein:

6. The absorption microscope probe of claim 5, wherein the micro microscope probe further comprises:

the liquid lens is positioned between the collimating lens and the micro-electromechanical scanning galvanometer to form a new first light path, and the new first light path sequentially comprises the collimating lens, the liquid lens, the micro-electromechanical scanning galvanometer, the first lens, the second lens, the dichroic mirror and the objective lens, wherein the collimating lens is positioned between the first port and the third port.

7. The absorption microscope detecting device according to claim 5, wherein a conducting fiber bundle is disposed around the outside of the objective lens of the micro microscope probe, and the conducting fiber bundle is used for conducting the optical signal before the objective lens to the dichroic mirror.

8. An absorption type three-dimensional nonlinear laser scanning microscope, comprising:

the multi-channel fluorescence collection device, the air extractor, the scanning collection controller, the femtosecond pulse laser, the fiber coupling module and the absorption microscope detection device according to any one of claims 5 to 7, wherein the multi-channel fluorescence collection device and the fiber coupling module are both connected with the absorption microscope detection device in an optical fiber communication manner, the multi-channel fluorescence collection device and the absorption microscope detection device are both electrically connected with the scanning collection controller, and the air extractor is electrically connected with the absorption microscope detection device, wherein:

the adsorption microscope 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 fluorescence signal and a second harmonic signal generated after the autofluorescent substance is excited through the objective lens, and outputting the fluorescence signal and the second harmonic signal to the multi-channel fluorescence collection device;

9. The absorption three-dimensional nonlinear laser scanning microscope according to claim 8, wherein the multi-channel fluorescence collection device comprises

the double-channel fluorescence collection module is used for collecting two-photon fluorescence signals and second harmonic signals by the probe of the micro microscope and synchronously converging the two-photon fluorescence signals and the second harmonic signals collected by the illumination multiplexing module to be converted into corresponding electric signals.

10. The absorption three-dimensional nonlinear laser scanning microscope according to claim 9, wherein the dual-channel fluorescence collection module comprises 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 collecting 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 fluorescent signals received by the multi-channel fluorescent collecting device, and the first photomultiplier is used for converting the fluorescent signals into first electric signals;

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

the first multiplexing collecting optical path sequentially comprises a lighting optical fiber bundle, a first multiplexing dichroic mirror, a first multiplexing collecting lens and a first transmission optical fiber.