CN210055991U

CN210055991U - Positioning type adsorption microscope detection device and laser scanning microscope

Info

Publication number: CN210055991U
Application number: CN201920180649.4U
Authority: CN
Inventors: 吴润龙; 王爱民; 王俊杰; 江文茂; 吴丹磊; 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: 2020-02-14
Anticipated expiration: 2029-01-31

Abstract

The embodiment of the utility model provides a locate mode adsorption microscope detecting device and laser scanning microscope. The positioning type adsorption microscope detection device comprises an adsorption shell and a micro microscope probe, wherein a movement device for driving the micro microscope probe to horizontally move along an X-Y axis is arranged on the inner side of the adsorption shell, a first light path for receiving and transmitting an output pulse laser signal to an autofluorescence substance in a cell of a living body, a second light path for collecting and transmitting a fluorescence signal and a second harmonic signal generated by exciting the autofluorescence substance and a zoom motor are arranged in the micro microscope probe. The embodiment of the utility model provides a locate mode adsorbs microscope detection device and laser scanning microscope and through miniaturized absorption casing, running gear and be provided with first light path, second light path and the miniature microscope probe of zooming motor, realizes surveying the accurate location of the three-dimensional different levels of the cell structure in the organism skin tissue, simple structure, convenient to use.

Description

Positioning type adsorption 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 adsorbs microscope detecting 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 adsorbs microscope detecting device and laser scanning microscope.

In a first aspect, an embodiment of the present invention provides a positioning type adsorption microscope detection device, include:

adsorb the casing and set up in adsorb miniature microscope probe in the casing, it is provided with the telecontrol equipment to adsorb the casing inboard, miniature microscope probe set up in telecontrol equipment is last, the telecontrol equipment is used for driving miniature microscope probe is along X-Y axle horizontal migration, wherein:

the telecontrol equipment with miniature microscope probe all set up in the adsorption housing, be provided with first light path, second light path and be used for the drive in the miniature microscope probe first light path with the shared infinity objective of second light path carries out the zoom motor that reciprocates, wherein:

the first optical path is used for receiving the pulse laser signal and transmitting and outputting the pulse laser signal to a spontaneous fluorescent substance in a cell of a living body;

the second optical path is used for collecting and transmitting a fluorescence signal and a second harmonic signal generated by the excitation of the autofluorescent substance.

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

fluorescence collection device, air exhaust device, scanning acquisition controller, femto second pulse laser instrument, fiber coupling module and the embodiment of the utility model provides a locate mode adsorbs microscope detecting device that the first aspect provided, fluorescence collection device with fiber coupling module all with locate mode adsorbs microscope detecting device optical fiber communication connection, fluorescence collection device with locate mode adsorbs microscope detecting device all with scanning acquisition controller electricity is connected, air exhaust device with locate mode adsorbs 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 collimating lens of the micro microscope probe in the positioning type adsorption microscope detection device;

the positioning type adsorption microscope detection device is used for receiving the pulse laser signal, outputting the pulse laser signal to an autofluorescent substance in a cell of a living body, 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 fluorescence collection device;

the 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 a scanning galvanometer in the probe of the miniature microscope to scan the pulse laser signals and synchronously acquiring the electric signals;

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

The embodiment of the utility model provides a locate mode adsorbs microscope detection device and laser scanning microscope includes adsorbing the casing and sets up the miniature microscope probe in adsorbing the casing, adsorbs adsorption type microscope detection device on the organism skin tissue of awaiting measuring through miniaturized adsorption casing to make miniature microscope probe survey the cell structure in the skin tissue, and through running gear X-Y axle horizontal migration adjustment, carries out the horizontal location of cell structure individual layer and surveys, and the zoom motor drive the common infinity objective of first light path with the second light path reciprocates, realizes the detection to the different degree of depth of the cell structure in the organism skin tissue, and then realizes the accurate location of the three-dimensional different levels of the cell structure in the organism skin tissue and surveys, and miniature microscope probe passes through built-in two light paths simultaneously, and the pulse laser signal output by the optical path excites the autofluorescence in the cell and obtains the fluorescence signal and the second harmonic signal generated by the autofluorescence, so that the three-dimensional laser scanning microscope provided with the adsorption type microscope detection device can carry out three-dimensional imaging on the cell structure through the obtained fluorescence signal and the second harmonic signal, and the three-dimensional laser scanning microscope has a simple structure and is convenient to use.

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 microscope detection device provided by an embodiment of the present invention;

fig. 2 is a schematic view of a cross-sectional structure of a combined positioning type absorption microscope detection device according to an embodiment of the present invention;

fig. 3 is a schematic view of a cross-sectional structure of a positioning type absorption microscope detection device according to another embodiment of the present invention;

fig. 4 is a schematic view of a cross-sectional structure of a positioning type absorption microscope detection device according to another embodiment of the present invention;

fig. 5 is a schematic sectional view of a positioning type absorption microscope detection device according to another embodiment of the present invention;

fig. 6 is a schematic structural view of an adsorption type three-dimensional nonlinear laser scanning microscope provided in an embodiment of the present invention;

fig. 7 is a schematic structural view of a fluorescence collecting device according to an embodiment of the present invention;

fig. 8 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. 9 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. 10 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. 11 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. 12 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. 13 is a schematic view of a box sealing structure of a box combination structure of an absorption 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 adsorbs microscope detecting device, fig. 1 does the embodiment of the utility model provides a locate mode adsorbs microscope detecting device structure schematic diagram, as shown in fig. 1, this locate mode adsorbs microscope detecting device includes:

adsorb casing 11 and set up miniature microscope probe 12 in adsorbing casing 11, adsorb casing 11 inboard and be provided with telecontrol equipment 13, miniature microscope probe 12 sets up on telecontrol equipment 13, telecontrol equipment 13 is used for driving miniature microscope probe 12 along X-Y axle horizontal migration, wherein:

the moving device 13 and the micro microscope probe 12 are both arranged in the adsorption shell 11, a first light path, a second light path and a zoom motor for driving the common infinite objective lens of the first light path and the second light path to move up and down are arranged in the micro microscope probe 12, wherein:

the first optical path is used for receiving the pulse laser signal and transmitting and outputting the pulse laser signal to the autofluorescence substance in the living body cell;

the second optical path is used for collecting and transmitting a fluorescent signal and a second harmonic signal generated by the excitation of the autofluorescent substance.

Specifically, the embodiment of the present invention provides a positioning type absorption microscope detecting device, which comprises an absorption casing 11 and a micro microscope probe 12 inside the absorption casing 11, wherein the absorption casing 11 is miniaturized to be absorbed on the surface of the skin of the living body, the micro microscope probe 12 inside the absorption casing 11 is used to detect the cell structure inside the skin of the living body, wherein the micro microscope probe 12 performs X-Y axis horizontal movement through an operation device disposed inside the absorption casing 11 to achieve horizontal positioning detection of the cell structure monolayer inside the skin of the living body, the zoom motor drives an infinite objective lens shared by a first light path and a second light path to move up and down, thereby achieving detection of different depths of the cell structure inside the skin of the living body, and further achieving precise positioning detection of three-dimensional different levels of the cell structure inside the skin of the living body, wherein, the micro microscope probe 12 is provided with a first light path for receiving the pulse laser signal and transmitting the output pulse laser signal to the autofluorescence substance in the living body cell and a second light path for collecting and transmitting the fluorescence signal and the second harmonic signal generated by the autofluorescence substance being excited, and the arrangement of the first light path and the second light path makes the adsorption type microscope detection device can complete the excitation of the autofluorescence substance in the living body skin cell and acquire the fluorescence signal and the second harmonic signal generated by the excitation and used for imaging the cell structure.

The embodiment of the utility model provides a locate mode adsorbs microscope detection device includes adsorbing the casing and sets up the miniature microscope probe in adsorbing the casing, adsorb adsorption type microscope detection device on the organism skin tissue of awaiting measuring through miniaturized adsorption casing, so that the miniature microscope probe surveys the cell structure in the skin tissue, and adjust through running device X-Y axle horizontal migration, carry out the horizontal location of cell structure individual layer and survey, zoom motor drive first light path and the common infinity objective of second light path reciprocate, realize the detection to the different degree of depth of cell structure in the organism skin tissue, and then realize the accurate location detection to the three-dimensional different levels of cell structure in the organism skin tissue, miniature microscope probe passes through built-in two light paths simultaneously, and the pulse laser signal through the light path output, the cell structure three-dimensional imaging device has the advantages that the autofluorescence substance in the cell is excited, and the fluorescence signal and the second harmonic signal generated by the autofluorescence substance are obtained, so that the three-dimensional laser scanning microscope provided with the adsorption type microscope detection device can perform three-dimensional imaging on the cell structure through the obtained fluorescence signal and the second harmonic signal, and the cell structure three-dimensional imaging device is simple in structure and convenient to use.

On the basis of each embodiment, the embodiment of the utility model provides an absorption casing among the locate mode absorption microscope detecting device includes shell body, base and cover glass, as shown in fig. 1, is provided with sucking disc 1121 on the base 112, and in sucking disc hole 1113 that the sucking disc 1121 embedding shell body 111 bottom was seted up, shell body 111 and base 112 passed through magnetic field force and can dismantle the connection, wherein:

the cover glass 113 is fixed to the sealing opening of the suction cup 1121 to form an internal space and an external suction space of the suction device, the moving device 13 and the micro microscope probe 12 are both disposed in the internal space, and the infinity objective is aligned with the cover glass 113 in the forward direction. That is, the outer shell 111 and the base 112 of the adsorption shell in the positioning type adsorption microscope detection device provided by the embodiment of the present invention are made of materials capable of being magnetically attracted to each other, or magnetic materials capable of being magnetically attracted to each other are embedded in the shell 111, so that the shell and the base can be detachably connected to each other through magnetic field force, and the base 112 is provided with the suction cup 1121, the bottom of the outer shell 111 is provided with the suction cup hole 1113 for embedding the suction cup 1121, the suction cup 1121 is provided with a sealing opening communicated with the space in the suction cup 1121, when the cover glass 113 covers the sealing opening, the adsorption microscope detection device forms an internal space and an external adsorption space capable of enabling the adsorption device to adsorb on the skin; and telecontrol equipment 13 and miniature microscope probe 12 all set up in built-in space, and miniature microscope probe 12 fixes the back, and infinity objective is positive to cover glass 113 to see through cover glass 113 and export internal signal and receive external signal, thereby realize that the microscope zooms, three-dimensional formation of image, and can dismantle the connection through magnetic field force between shell body 111 and the base 112, make things convenient for the staff just can change cover glass 113 through dismantling base 112, easy operation, convenient to use.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides a first light path among the locate mode adsorption microscope detecting device is including the first light, second light and the third light that meet in proper order, wherein:

the pulse laser signal transmits the collimating lens, the analyzer, the polarization beam splitter, the first quarter-wave plate and the scanning galvanometer in the first light path to form first light;

after the first light is formed, the pulse laser signal is reflected by the scanning galvanometer and then transmits the first quarter-wave plate to the polarization spectroscope again to form second light;

after the second light is formed, the pulse laser signal is reflected by the polarizing beam splitter and then transmits the first scanning mirror, the second scanning mirror and the dichroic mirror to the infinite objective lens to form third light;

the second optical path comprises an infinite objective lens and a dichroic mirror which are used for collecting and transmitting a fluorescent signal and a second harmonic signal in sequence. That is, fig. 2 is a schematic view of a cross-sectional structure of the positioning type absorption microscope detection device after combination according to an embodiment of the present invention, wherein a light path in a micro microscope probe is involved, as shown in fig. 2, when a pulse laser signal is processed by a first light path in the positioning type absorption microscope detection device provided by an embodiment of the present invention, the pulse laser signal can form three light beams due to the light path turning back when passing through an optical device of the first light path, the three light beams are respectively a first light beam, a second light beam and a third light beam, a collimating lens 121, an analyzer 122, a polarization beam splitter 123, a first quarter wave plate 124 to a scanning beam splitter 125 are arranged in the light path forming the first light beam, a scanning beam splitter 125, a first quarter wave plate 124 and a polarization beam splitter 123 are arranged in the light path forming the second light beam, a polarization beam splitter 123, a first transmission scanning mirror 126 are arranged in the light path forming the third light beam, A second scanning mirror 127, a dichroic mirror 128, and an infinity objective 129;

after the pulse laser signal exits from the optical fiber, the pulse laser signal enters the analyzer 122 after being collimated by the collimating lens 121, so that collimated light is changed into linearly polarized light, the polarization direction of the linearly polarized light is consistent with the transmission polarization direction of the polarization beam splitter 123, therefore, the linearly polarized light enters the polarization beam splitter 123, directly transmits through the polarization beam splitter 123, enters the first quarter-wave plate 124 positioned on the left side of the polarization beam splitter 123, the fast axis direction of the first quarter-wave plate forms an angle of +/-45 degrees with the polarization direction of the linearly polarized light, and after passing through the first quarter-wave plate 124, the linearly polarized light is changed into circularly polarized light and is incident on the scanning polarizer 125 to form the first light; after the first light is formed, the circularly polarized light reflected by the scanning galvanometer 125 enters the first quarter-wave plate 124 again to be changed into linearly polarized light, wherein the polarization direction of the linearly polarized light is perpendicular to the transmission polarization direction of the polarizing beam splitter 123, so that the linearly polarized light is reflected on the splitting surface of the polarizing beam splitter 123 and exits from the lower part of the polarizing beam splitter 123 to form a second light; after the second light is formed, the emergent collimated light enters the scanning mirror to be converged and enters the infinite objective 129, and the focal point of the infinite objective is located at the relay image surface to form a third light; scanning the mirror 125 in the XY axis sweeps the pulsed laser signal focus over the relay image plane. The pulse laser signal is transmitted through a parallel plate as a dichroic mirror 128, and the dichroic mirror 128 distinguishes the pulse laser signal and a fluorescent signal and a second harmonic signal generated by excitation of an autofluorescent substance according to the wavelength, transmits the pulse laser signal therethrough, and reflects the fluorescent signal and the second harmonic signal. The relay image plane generated by the pulse laser signal converged by the scanning mirror coincides with the rear image plane of the infinity objective 129. The relay image produced by the pulsed laser signal scan is thus scaled onto the sample according to the magnification of the infinity objective 129. The focus of the pulse laser signal on the sample can generate a fluorescence signal and a second harmonic signal, the generated fluorescence signal and the second harmonic signal are collected by the infinite objective 129 and reflected by the dichroic mirror 128 to enter the collection optical fiber bundle for collection, wherein the zoom motor 15 drives the infinite objective 129 to move up and down; wherein the scanning galvanometer may be based on mechanical, electromechanical or microelectromechanical principles, as is the case with the various embodiments below.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides a first light path among the locate mode adsorption microscope detecting device is including meeting first light, second light, third light, fourth light and fifth light in proper order, wherein:

the pulse laser signal is incident to the polarization spectroscope through a collimating lens and an analyzer in a first light path to form first light;

after the first light is formed, the pulse laser signal is reflected by the polarization beam splitter and then transmits the second quarter wave plate to the plane reflector to form second light;

after the second light is formed, the pulse laser signal is reflected by the plane mirror and then transmits the second quarter wave plate, the polarization beam splitter and the third quarter wave plate to the scanning galvanometer again to form third light;

after the third light is formed, the pulse laser signal is reflected by the scanning galvanometer and then transmits the third quarter-wave plate to the polarization spectroscope again to form a fourth light;

after the fourth light is formed, the pulse laser signal is reflected by the polarizing beam splitter and then transmits the first scanning mirror, the second scanning mirror and the dichroic mirror to the infinity objective lens to form a fifth light;

the second optical path comprises an infinite objective lens and a dichroic mirror which are used for collecting and transmitting a fluorescent signal and a second harmonic signal in sequence. That is, fig. 3 is a schematic view of a cross-sectional structure of the positioning type absorption microscope detection device after combination according to another embodiment of the present invention, wherein the cross-sectional structure relates to a light path in a micro microscope probe, as shown in fig. 3, when a pulse laser signal is processed by a first light path in the positioning type absorption microscope detection device provided by the embodiment of the present invention, the pulse laser signal can form five light beams when passing through an optical device of the first light path due to the turn-back of the light path, the light path includes a first light beam, a second light beam, a third light beam, a fourth light beam and a fifth light beam which are sequentially connected, a collimating lens 140, an analyzer 141 and a polarization beam splitter 142 are arranged in the light path forming the first light beam, the polarization beam splitter 142, a second quarter wave plate 143 and a plane reflector 144 are arranged in the light path forming the second light beam, and a plane reflector 144 is arranged in the light path forming the, The second quarter-wave plate 143, the polarization beam splitter 142, the third quarter-wave plate 145 and the scanning galvanometer 146 are arranged in the light path forming the fourth light ray, the scanning galvanometer 146, the third quarter-wave plate 145 and the polarization beam splitter 142 are arranged in the light path forming the fifth light ray, and the polarization beam splitter 142, the first scanning mirror 147, the first scanning mirror 148, the dichroic mirror 149 and the infinity objective 160 are arranged in the light path forming the fifth light ray;

after the pulse laser signal exits from the optical fiber, the pulse laser signal enters the analyzer 141 after being collimated by the collimating lens 140, so that the collimated light is changed into linearly polarized light, the polarization direction of the linearly polarized light is perpendicular to the transmission polarization direction of the polarization beam splitter 142, and the linearly polarized light enters the polarization beam splitter 142, is reflected and exits from the right side of the polarization beam splitter 142, so that a first light ray is formed; after the first light is formed, the emitted linearly polarized light enters the second quarter-wave plate 143, wherein the fast axis direction of the second quarter-wave plate forms an angle of ± 45 degrees with the linearly polarized light polarization direction, so that the linearly polarized light is changed into circularly polarized light after passing through the second quarter-wave plate and is incident to the plane mirror 144 to form a second light; after the second light is formed, the circularly polarized light is reflected by the plane mirror 144, returns along the original optical path, and is changed into linearly polarized light by the second quarter-wave plate 143 again, and the polarization direction of the linearly polarized light is consistent with the transmission polarization direction of the polarization beam splitter 142, so that the linearly polarized light enters the polarization beam splitter 142 for the second time and then directly transmits through the polarization beam splitter 142, enters the third quarter-wave plate 145 positioned on the left side of the polarization beam splitter 142, wherein the fast axis direction of the third quarter-wave plate forms an angle of ± 45 degrees with the polarization direction of the linearly polarized light, and the linearly polarized light is changed into circularly polarized light after passing through the third quarter-wave plate and then enters the scanning polarizer 146; after the third light is formed, the circularly polarized light reflected by the scanning galvanometer 146 enters the third quarter-wave plate 145 again to be changed into linearly polarized light, wherein the polarization direction of the linearly polarized light is perpendicular to the transmission polarization direction of the polarization beam splitter 142, so that the polarized light is reflected on the splitting surface of the polarization beam splitter 142 and exits from the lower part of the polarization beam splitter 142 to form a fourth light; after the fourth light is formed, the emergent collimated light enters the scanning mirror to be converged and enters the infinite objective lens 160, and the focal point of the infinite objective lens is located at the relay image surface to form a fifth light; scanning the mirror 146 in the XY axis sweeps the pulsed laser signal focal point across the relay image plane. The pulse laser signal is transmitted through a parallel plate as a dichroic mirror 149, and the dichroic mirror 149 distinguishes the pulse laser signal and a fluorescent signal and a second harmonic signal generated by excitation of an autofluorescent substance according to the wavelength, transmits the pulse laser signal therethrough, and reflects the fluorescent signal and the second harmonic signal. The relay image plane generated by the pulse laser signal converged by the scanning mirror coincides with the rear image plane of the infinity objective 160. The relay image produced by the pulsed laser signal scan is thus scaled onto the sample according to the magnification of the infinity objective 160. The focus of the pulsed laser signal on the sample generates a fluorescence signal and a second harmonic signal, which are collected by the infinity objective 160 and reflected by the dichroic mirror 149 into the collection fiber bundle.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides an absorption formula microscope detection device still includes the adjustable curvature lens of electricity, and fig. 4 is the utility model discloses the section structure sketch map after the microscope detection device combination is adsorbed to locate mode that still another embodiment provided, as shown in fig. 4, the adjustable curvature lens of electricity 120 is located between collimating lens 121 and the analyzer 122, and pulse laser signal transmits collimating lens 121, the adjustable curvature lens of electricity 120, analyzer 122, polarization spectroscope 123, first quarter wave plate 124 to scanning galvanometer 125, forms new first light. That is, the optical elements in the new first light ray sequentially include a collimating lens 121, an electrically adjustable curvature lens 120, an analyzer 122, a polarization beam splitter 123, a first quarter-wave plate 124 and a scanning galvanometer 125. The electrically adjustable curvature lens 120 is disposed such that a corresponding curvature of the surface of the electrically adjustable curvature lens 120 can be generated by applying a voltage or a current to the electrically adjustable curvature lens 120, and thus parallel light emitted from the straight lens 121 can be aligned to generate different focal powers. The specific light path is as follows: laser signals are emitted from the optical fibers, parallelly enter the electric adjustable curvature lens 120 after passing through the collimating lens 121, corresponding focal power is generated from the electric adjustable curvature lens 120 according to loaded voltage or current signals, and emergent convergent or divergent light passes through an optical element in a first light path to form new first light, second light and third light which are transmitted to the infinity objective 129 and then converged on a sample. The focal power change introduced by the electrically adjustable curvature lens 120 can make the focal point of the laser signal emitted from the infinite objective 129 port move up and down in the depth direction, and the electrically adjustable curvature lens 120 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 electrically adjustable curvature lens 120 is equivalent to parallel plate glass when no voltage or current signal is applied, and has no focal power to the laser signal and does not cause any shift of the focus behind the infinity objective 129, thereby realizing three-dimensional stereoscopic imaging. When the system is used specifically, the electrically adjustable curvature lens 120 is complementary to the zoom motor 13, the position of the infinite objective 129 is adjusted through the zoom motor 13, after the system is roughly adjusted to the corresponding depth position, the system is switched to the zoom scanning mode of the electrically adjustable curvature lens 120, and a sample is rapidly three-dimensionally imaged, wherein when the adsorption type microscope detection device is not provided with the zoom motor 13, the zoom adjustment can be performed only through the electrically adjustable curvature lens 120.

On the basis of each embodiment, the embodiment of the utility model provides an absorption formula microscope detection device still includes the adjustable curvature lens of electricity, and fig. 5 is the utility model discloses the section structure sketch map after the microscope detection device combination is adsorbed to locate mode that still another embodiment provided, as shown in fig. 5, the adjustable curvature lens of electricity 150 is located between collimating lens 140 and the analyzer 141, and pulse laser signal transmits collimating lens 140, the adjustable curvature lens of electricity 150, the analyzer 141 and incides polarization spectroscope 142, forms new first light. That is, the optical elements in the new first light ray sequentially include a collimating lens 140, an electrically adjustable curvature lens 150, an analyzer 141, and a polarizing beam splitter 142. The electrically adjustable curvature lens 150 is configured such that a voltage or a current is applied to the electrically adjustable curvature lens 150 to cause a corresponding curvature of the surface of the electrically adjustable curvature lens 150, thereby generating different powers for the parallel light emitted from the collimating lens 140. The specific light path is as follows: laser signals are emitted from the optical fibers, parallelly enter the electric adjustable curvature lens 150 after passing through the collimating lens 140, corresponding focal power is generated from the electric adjustable curvature lens 150 according to loaded voltage or current signals, and the emitted convergent or divergent light passes through the optical element in the first light path to form new first light, second light, third light, fourth light and fifth light, and then the new first light, second light, third light, fourth light and fifth light are transmitted to the infinity objective 160 and converged on a sample. The focal power change introduced by the electrically adjustable curvature lens 150 can make the focal point of the laser signal emitted from the infinity objective 160 port move up and down in the depth direction, and the electrically adjustable curvature lens 150 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 electrically adjustable curvature lens 150 is equivalent to parallel plate glass when no voltage or current signal is applied, and has no focal power to the laser signal and does not cause any shift of the focus behind the infinity objective 160, thereby realizing three-dimensional stereoscopic imaging. When the system is used specifically, the electrically adjustable curvature lens 150 is complementary to a zoom motor, the position of the infinite objective lens 160 is adjusted by the zoom motor, after the system is roughly adjusted to the corresponding depth position, the system is switched to the zoom scanning mode of the electrically adjustable curvature lens 150, and a sample is rapidly three-dimensionally imaged, wherein when the adsorption type microscope detection device is not provided with the zoom motor, the zoom adjustment can be performed only by the electrically adjustable curvature lens 150.

On the basis of each embodiment, the embodiment of the utility model provides a kinematic device in locate mode adsorption microscope detection device includes fixed bolster, first drive block and second drive block, as shown in fig. 1, first drive block 132 and second drive block 133 respectively with fixed bolster 131 fixed connection, second drive block 133 is fixed on the lateral wall of adsorbing casing 11, wherein:

the second driving block 133 is used to drive the first driving block 132 to move in the Y-axis direction together with the fixing bracket 131, and the first driving block 132 is used to drive the micro microscope probe to move in the X-axis direction. On the basis of the above embodiments, the moving device in the positioning type absorption microscope detection device provided by the embodiment of the present invention includes a fixing bracket 131, a first driving block 132 and a second driving block 133, the first driving block 132 and the second driving block 133 both include a sliding block and a fixing block, the first fixing block of the first driving block 132 is fixed on the fixing bracket 131, and the first sliding block of the first driving block 132 slides relative to the first fixing block under the driving of the first motor; the second slider of the second driving block 133 is fixed on the fixed bracket 131, the second fixed block of the second driving block 133 is fixed on the side wall of the outer shell 11, and the second slider of the second driving block 133 slides relative to the second fixed block under the driving of the second motor; the micro microscope probe 12 can be fixed on the first slider of the first driving block 132, so that the first driving block 132 can drive the micro microscope probe 12 to move along the X-axis direction, the second driving block 133 can drive the first driving block 132 and the fixing bracket 131 and the micro microscope probe 12 fixed on the first driving block 132 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 12 can be driven to move along the X-axis and the Y-axis directions by the movement device, so as to realize the accurate positioning of the micro microscope probe 12 in the horizontal plane, and meanwhile, the micro microscope probe 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 step 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.

On the basis of each embodiment, the embodiment of the utility model provides a fixed bolster in positioning type adsorption microscope detecting device includes X-axis support and Y-axis support, X-axis support and Y-axis support mutually perpendicular fixed connection, 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 the embodiment of the utility model provides a fixed bolster among location formula adsorption microscope detecting device 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 the X-axis support, and fixed connection can be dismantled with the second slider of second drive block to the realization miniature microscope probe can be followed XY axle direction and removed under telecontrol equipment's drive.

On the basis of the above embodiments, the sucker in the positioning type suction device for setting the micro microscope probe provided by the embodiment of the utility model further comprises a suction port and an air suction port, wherein the suction port is communicated with the sealing port and is used for being sucked on the life body to be measured through the external suction space;

the air pumping port is communicated with the outer adsorption space and is used for pumping the air in the outer adsorption space. That is, in the positioning type adsorption microscope detection device provided by the embodiment of the present invention, the suction cup disposed on the base includes an adsorption port and an air extraction port in addition to the sealing port, and the sealing port is communicated with the adsorption port to form an internal space of the suction cup, when the cover glass is sealed and fixed on the sealing port, the internal space of the suction cup forms an external adsorption space, and the adsorption port is adsorbed on the life body to be detected through the external adsorption space; the air suction port is communicated with the outer adsorption space, and air in the outer adsorption space can be sucked through the air suction port, so that negative pressure in the outer adsorption space is formed. Under the action of external pressure, the positioning type adsorption microscope detection device is adsorbed on tissues such as skin of a to-be-detected living body.

On the basis of the above embodiments, the embodiment of the present invention provides an outer casing in a positioning type absorption device for setting a micro microscope probe, including a first casing and a second casing, as shown in fig. 1, wherein:

an accommodating space is arranged in the first housing 1111, the movement device is arranged in the accommodating space, and the first housing 1111 and the second housing 1112 are detachably and fixedly connected. That is, the embodiment of the present invention provides an outer casing in a positioning type absorption device for setting a probe of a micro microscope, which comprises two parts, namely a first casing 1111 and a second casing 1112, wherein the first casing 1111 has a containing space for setting a motion device, that is, the sucker hole in the above embodiment is also arranged at the bottom of the first casing 1111, the sucker is embedded into the sucker hole of the first casing 1111, and then the cover glass and the second casing 1112 are combined to form an external absorption space and an internal space in the above embodiment, wherein the first casing 1111 and the second casing 1112 can be detachably fixed by screws, so as to facilitate the assembly, disassembly and replacement of parts of the whole device.

The embodiment of the utility model provides a still provide an absorption formula three-dimensional nonlinear laser scanning microscope, fig. 6 is the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope structure schematic diagram, as shown in fig. 6, this laser scanning microscope includes:

fluorescence collection device 56, air exhaust device 52, scanning acquisition controller 531, femtosecond pulse laser 541, optical fiber coupling module 542 and the locate mode absorption microscope detecting device 51 that the above-mentioned embodiment provided, fluorescence collection device 56 and optical fiber coupling module 542 all with locate mode absorption microscope detecting device 51 optical fiber communication connection, fluorescence collection device 56 and locate mode absorption microscope detecting device 51 all are connected with scanning acquisition controller 531 electricity, air exhaust device 52 is connected with locate mode absorption microscope detecting device 51 electricity, wherein:

the femtosecond pulse laser 541 is used for outputting a pulse laser signal to the optical fiber coupling module 542;

the optical fiber coupling module 542 is used for coupling the pulse laser signal output by the femtosecond pulse laser 541 and transmitting the pulse laser signal to a micro microscope probe in the positioning type adsorption microscope detection device;

the positioning type adsorption microscope detection device is used for receiving the pulse laser signal, outputting the pulse laser signal to the autofluorescence substance in the living body cell, acquiring a fluorescence signal and a second harmonic signal generated after the autofluorescence substance is excited through the objective lens, and outputting the fluorescence signal and the second harmonic signal to the fluorescence collection device 56;

a fluorescence collecting device 56 for receiving the fluorescence signal and the second harmonic signal and converting the fluorescence signal and the second harmonic signal into corresponding electrical signals;

the scanning acquisition controller 531 is used for controlling a scanning galvanometer in the probe of the miniature microscope to scan the pulse laser signals and synchronously acquire electric signals;

and the air exhausting device 52 is used for exhausting the outer adsorption space of the positioning type adsorption microscope detection device so as to form negative pressure in the outer adsorption space.

Specifically, the embodiment of the present invention provides an adsorption type three-dimensional nonlinear laser scanning microscope, which includes a fluorescence collecting device 56, an air extractor 52, a scan collecting controller 531, a femtosecond pulse laser 541, an optical fiber coupling module 542 and a positioning type adsorption microscope detecting device 51, so as to form a three-dimensional nonlinear laser scanning microscope capable of being adsorbed on human skin or detecting in deep into human intestines and stomach, wherein the femtosecond pulse laser 541 can emit pulse laser signals for exciting autofluorescence in human skin cells, generating multiphoton fluorescence signals and second harmonic signals, including using the 920nm femtosecond pulse laser 541 to excite FAD and collagen in cells, exciting the 500-, to generate corresponding fluorescent and second harmonic signals;

the fluorescence collecting device 56 integrates two signal collecting optical paths, namely a fluorescence signal collecting optical path and a second harmonic signal collecting optical path, to respectively collect the fluorescence signal and the second harmonic signal; the scanning acquisition controller 531 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 fluorescent collection device 56; the air extracting device 52 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 above embodiment, an air extracting valve is arranged in the air extracting pipeline, the air extracting valve is electrically connected with the air extracting device 52, the air extracting device 52 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 13 frames (512 pixels by 512 pixels).

The embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope adopts fluorescence collection device, air exhaust device, scanning acquisition controller, femto second pulse laser, optical fiber coupling module and locate mode absorption microscope detection device, thereby form can adsorb the three-dimensional nonlinear laser scanning microscope that can adsorb and go deep into human intestines and stomach and survey, 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 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 locate mode adsorption microscope detecting 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 that multiple imaging mode includes 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 3D image of reconsitution, the equipment operation is simple, high durability and convenient use.

On the basis of the foregoing embodiments, fig. 7 is a schematic structural diagram of the fluorescence collecting device provided by the embodiment of the present invention, as shown in fig. 7, the embodiment of the present invention provides a fluorescence collecting device including an optical fiber universal interface 881, a first photomultiplier 882, a second photomultiplier 883, a first collecting optical path between the optical fiber universal interface 881 and the first photomultiplier 882, and a second collecting optical path between the optical fiber universal interface 881 and the second photomultiplier 883, wherein:

the first collecting light path sequentially comprises a coupling collecting 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 includes a coupling collecting 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 configured to collect a second harmonic signal received by the fluorescence collecting device, and the second photomultiplier 883 is configured to convert the second harmonic signal into a second electrical signal. That is, the fluorescence collecting device provided by the embodiment of the present invention has a two-way signal collecting function, and integrates two light paths, wherein the first dichroic mirror 83 in the first collecting light path is a transmission fluorescence signal, the dichroic mirror for reflecting the second harmonic, the second dichroic mirror 86 and the first dichroic mirror 83 are the same dichroic mirror, and are used for reflecting the second harmonic, the first optical filter 84 is used for transmitting the fluorescence signal, and filtering the rest interference signals, and the second optical filter 87 is used for transmitting the corresponding second harmonic signal and filtering the rest interference signals, for example, when using 880nm femtosecond fiber laser to excite the autofluorescence substance in the human body surface skin cell, 390nm second harmonic signal and 450 and 600nm two-photon autofluorescence signal can be obtained, the wavelength passes through above 420nm, the dichroic mirror reflected by the wavelength below 420, that is, the first dichroic mirror 83 can separate the two-way fluorescence, clean second harmonic signals and fluorescence signals can be obtained by using the first filter 84 of 390 + -20 nm and the second filter 87 of 450-600nm, respectively.

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 industrial computer, as shown in FIG. 6, industrial computer 532 is connected with scanning acquisition controller 531 electricity, wherein:

the industrial personal computer 532 is configured to obtain the first electrical signal and the second electrical signal acquired by the scanning acquisition controller 531, generate a first fluorescent image based on the first electrical signal, and generate a second fluorescent image based on the second electrical signal. That is to say the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes the industrial computer 532 with scan acquisition controller 531 electricity is connected, this industrial computer 532 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 install control software on the industrial computer, through control software, send control command to the scanner to control scan acquisition controller, acquire above-mentioned first signal of telecommunication and second signal of telecommunication.

On the basis of each embodiment, the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes the display, as shown in fig. 6, display 55 is connected with industrial computer 532 electricity for show first fluorescence image and second fluorescence image. That is, the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope still includes the display 55 that is used for showing first fluorescence image and second fluorescence image, through display 55, the staff can directly acquire first fluorescence image and second fluorescence image's relevant information.

On the basis of above-mentioned each embodiment, the embodiment of the utility model provides a locate mode among the absorption formula three-dimensional nonlinear laser scanning microscope detection device is a plurality ofly. Namely the embodiment of the utility model provides a fluorescence collection device and fiber coupling module can adsorb microscope detection device fiber communication connection with a plurality of locate mode simultaneously, 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 fluorescence collection device and fiber coupling module respectively with the locate mode adsorb the optic fibre transmission connection between the microscope detection device, wherein:

the length of the adjusting optical fiber can be adjusted. Namely the embodiment of the utility model provides a fluorescence collection device and optical fiber coupling module among absorption formula three-dimensional nonlinear laser scanning microscope adsorb microscope and carry out optical fiber transmission through adjustable length's regulation optic fibre and locate mode respectively and be 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 to explain, fig. 8 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. 8, the positioning type adsorption microscope detection 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 fluorescence collection device, the optical fiber coupling module and the fluorescence collection device are both in optical fiber transmission connection with the positioning type adsorption microscope detection device 51, wherein, the working principle of the adsorption type three-dimensional nonlinear laser scanning microscope is the same as that of the, and will not be described in detail herein.

Wherein, fig. 9 is the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope surveys human chest skin tissue schematic diagram, as shown in 9, through air exhaust device 52's air exhaust function, adsorb microscope detection device 51 at human chest with the locate mode, 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, optic fibre coupling module and fluorescence collection device all adsorb microscope detection device 51 optical fiber transmission with the locate mode and be connected, 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. 10 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. 10, a plurality of positioning type absorption 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 fluorescence collection device are integrated in a second device 54, the optical fiber coupling module and the fluorescence collection device are both in optical fiber transmission connection with the positioning type absorption microscope detection devices 51, thereby, the skin tissue structures of different parts of the human body can be simultaneously detected under the effect of the plurality of positioning type absorption microscope detection devices, the operation is, and because the positioning type adsorption microscope detection device is connected with the second device through 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. 11 is the embodiment of the utility model provides an absorption formula three-dimensional nonlinear laser scanning microscope surveys animal skin tissue sketch map, as shown in fig. 11, equally can be through air exhaust device 52's air exhaust function, adsorb microscope detecting device 51 on the skin tissue of life entity with the locate mode, 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 fluorescence collection device all with locate mode absorption microscope detecting 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. 12 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. 12, this three-dimensional nonlinear laser scanning microscope's of absorption formula scanning microscope scanning acquisition controller 531, industrial computer 532, air exhaust device 52, fluorescence collection device 56 and femto second pulse laser and fiber coupling integrated module 540 together, integrated 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 positioning type adsorption microscope detection device 51 is adsorbed on the skin tissue of a human body to be detected, is in optical fiber communication connection with the optical fiber coupling and fluorescence collection device 56 in the box body, is connected with the air pump through an air extraction pipeline, and is electrically connected with the power plug, the scanning acquisition controller 531 and the industrial personal computer 532. Wherein fig. 13 is the utility model provides an adsorption type three-dimensional nonlinear laser scanning microscope's box integrated configuration's joint sealing structure sketch map, as shown in fig. 13, display 55 is integrated together 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 microscope detection device is characterized by comprising:

2. The positioning type adsorption microscope detection device according to claim 1, wherein the adsorption casing comprises an outer casing, a base and a cover glass, the base is provided with a suction cup, the suction cup is embedded in a suction cup hole formed in the bottom of the outer casing, the outer casing is detachably connected with the base through magnetic field force, and the positioning type adsorption microscope detection device comprises:

the cover glass is fixed on the sealing opening of the sucker to form a built-in space and an external adsorption space of the positioning type adsorption microscope detection device, the motion device and the miniature microscope probe are arranged in the built-in space, and the infinite objective is aligned to the cover glass in the forward direction.

3. The apparatus according to claim 1, wherein the first light path comprises a first light beam, a second light beam and a third light beam, which are connected in sequence, wherein:

after the second light is formed, the pulse laser signal is reflected by the polarizing beam splitter and then transmits through the first scanning mirror, the second scanning mirror and the dichroic mirror to the infinity objective lens to form a third light;

the second optical path sequentially comprises the infinity objective and the dichroic mirror which collect and transmit the fluorescence signal and the second harmonic signal.

4. The apparatus according to claim 1, wherein the first light path comprises a first light beam, a second light beam, a third light beam, a fourth light beam and a fifth light beam, which are connected in sequence, wherein:

the pulse laser signal is incident to a polarization spectroscope through a collimating lens and an analyzer in the first light path to form first light;

after the first light is formed, the pulse laser signal is reflected by the polarization beam splitter and then transmits a second quarter wave plate to the plane reflector to form a second light;

after the second light is formed, the pulse laser signal is reflected by the plane mirror and then transmits the second quarter wave plate, the polarization beam splitter and the third quarter wave plate to the scanning galvanometer again to form a third light;

after the third light is formed, the pulse laser signal is reflected by the scanning galvanometer and then transmits the third quarter wave plate to the polarization spectroscope again to form fourth light;

after the fourth light is formed, the pulse laser signal is reflected by the polarizing beam splitter and then transmits through the first scanning mirror, the second scanning mirror and the dichroic mirror to the infinity objective lens to form a fifth light;

the second optical path sequentially comprises the infinity objective and the dichroic mirror which collect and conduct the fluorescence signal and the second harmonic signal.

5. The apparatus according to claim 3, further comprising an electrically tunable curvature lens disposed between the collimating lens and the analyzer, wherein the pulsed laser signal transmits through the collimating lens, the electrically tunable curvature lens, the analyzer, the polarizing beam splitter, the first quarter wave plate, and the scanning galvanometer to form a new first light beam.

6. The apparatus according to claim 4, further comprising an electrically adjustable curvature lens disposed between the collimating lens and the analyzer, wherein the pulsed laser signal is transmitted through the collimating lens, the electrically adjustable curvature lens, and the analyzer to enter the polarizing beam splitter to form a new first light beam.

7. The apparatus according to claim 1, wherein the moving device comprises a fixed frame, a first driving block and a second driving block, the first driving block and the second driving block are respectively fixed to the fixed frame, and the second driving block is fixed to a sidewall of the adsorption housing, wherein:

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.

8. The apparatus according to claim 7, wherein the fixed frame comprises an X-axis frame and a Y-axis frame, the X-axis frame and the Y-axis frame are vertically and fixedly connected to each other, 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.

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

the apparatus comprises a fluorescence collection device, an air extraction device, a scanning collection controller, a femtosecond pulse laser, an optical fiber coupling module and the positioning type adsorption microscope detection device as claimed in any one of claims 1 to 8, wherein the fluorescence collection device and the optical fiber coupling module are both connected with the positioning type adsorption microscope detection device in an optical fiber communication manner, the fluorescence collection device and the positioning type adsorption microscope detection device are both electrically connected with the scanning collection controller, the air extraction device is electrically connected with the positioning type adsorption microscope detection device, and the apparatus comprises:

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 micro microscope probe in the positioning type adsorption microscope detection device;

10. The absorption three-dimensional nonlinear laser scanning microscope according to claim 9, wherein the fluorescence collecting device comprises a fiber-optic universal interface, a first photomultiplier tube, a second photomultiplier tube, and a first collecting optical path between the fiber-optic universal interface and the first photomultiplier tube and a second collecting 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 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, the second filter and the second collecting lens, wherein the second collecting light path is used for collecting the second harmonic signals received by the fluorescence collecting device, and the second photomultiplier is used for converting the second harmonic signals into second electric signals.