CN109745007A - Positioning formula adsorbent equipment, microscope detection device and laser scanning microscope - Google Patents
Positioning formula adsorbent equipment, microscope detection device and laser scanning microscope Download PDFInfo
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- CN109745007A CN109745007A CN201910099549.3A CN201910099549A CN109745007A CN 109745007 A CN109745007 A CN 109745007A CN 201910099549 A CN201910099549 A CN 201910099549A CN 109745007 A CN109745007 A CN 109745007A
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Abstract
The embodiment of the present invention provides a kind of positioning formula adsorbent equipment, microscope detection device and laser scanning microscope.Wherein, above-mentioned positioning formula adsorbent equipment includes outer housing, pedestal, coverslip and telecontrol equipment, and outer housing and pedestal are detachably connected by magnetic field force, and the seal that coverslip is fixed on sucker forms built-in space and outer adsorption space；Telecontrol equipment includes fixed bracket, the first drive block and the second drive block, and the second drive block drives the first drive block and fixed bracket to move along the y axis, and the first drive block driving mini microscope probe moves along the x axis.Positioning formula adsorbent equipment, microscope detection device and laser scanning microscope provided in an embodiment of the present invention pass through the first drive block and the second drive block, realize that mini microscope probe is moved along XY axis direction in the horizontal plane, in the case where not moving adsorbent equipment, to find point of observation, realize the accurate positioning of mini microscope probe, it is easy to operate, easy to use.
Description
Technical field
The present embodiments relate to laser scanning microscope technical field more particularly to a kind of positioning formula adsorbent equipment, show
Micro mirror detection device and laser scanning microscope.
Background technique
With the continuous development of medicine and biology, people are to cellular morphology, institutional framework or stomach in animal life
The research of middle fiber condition achieves marked improvement, particularly by the pulsed laser radiation excitation of near infrared region and by suitable
The detection of high sensitivity receiver, fluorescence signal and second harmonic signal are obtained, to obtain the biological cell form of living body
The relevant technologies achieve significant achievement.
And it is based on fluorescence signal, second harmonic signal and CARS (Coherent anti-Stokes Raman
Scattering, related anti-Stokes Raman scattering) signal, to obtain the correlation detection equipment of biological cell form, upper
It states and is play an important role in the application of technology.The existing imaging device for being used for human body cell or tissue detection, it is mainly three-dimensional
Non-linear laser flying-spot microscope, wherein the form of current above-mentioned laser scanning microscope is predominantly moved by mechanical arm
The detection device installation of the i.e. laser scanning microscope of the laser scanning microscope of microscope detection device on the robotic arm, passes through
The adjustment of mechanical arm carrys out movement detector, is then aligned with the different institutional framework of detection human body.
But the detection device based on mechanical arm in above-mentioned three dimensional non-linear laser scanning microscope, due to its volume compared with
Greatly, it pops one's head in the corresponding biggish skin area of human body, it, can not be into for the specific position of skin detection so that in concrete operations
Row is accurately positioned, and needs repeatedly to adjust mechanical arm, and multiple movement detector, locating effect is poor, is easy to produce offset deviation, from
And influence image quality.
Summary of the invention
For the technical problems in the prior art, the embodiment of the present invention provides a kind of positioning formula adsorbent equipment, micro-
Mirror detection device and laser scanning microscope.
In a first aspect, the embodiment of the present invention provides a kind of positioning formula adsorbent equipment popped one's head in for mini microscope to be arranged,
Include:
Outer housing, pedestal, coverslip and the telecontrol equipment for driving mini microscope probe to move in the horizontal plane, it is described
Sucker is provided on pedestal, the sucker is embedded in the sucker hole that the outer housing bottom opens up, the outer housing and the bottom
Seat is detachably connected by magnetic field force, in which:
The coverslip is fixed on the seal of the sucker, formed the adsorption device built-in space and outer absorption
Space；
The telecontrol equipment is set in the built-in space, and the telecontrol equipment includes fixed bracket, the first drive block
With the second drive block, first drive block and second drive block are fixedly connected with the fixed bracket respectively, and described
Two drive blocks are fixed on the side wall of the outer housing, in which:
Second drive block is used to drive first drive block and the fixed bracket mobile in Y direction together,
First drive block is mobile in X-direction for driving mini microscope probe, wherein mini microscope probe is positive
It is directed at the coverslip.
Second aspect, the embodiment of the present invention provide a kind of absorption type microscope detection device, comprising:
What mini microscope probe and first aspect of the embodiment of the present invention provided is used to be arranged mini microscope probe
Positioning formula adsorbent equipment, the mini microscope probe are fixed on first drive block, in which:
The mini microscope probe includes probing shell and fixed plate, and it is logical that the probing shell top is provided with first
Mouth and the second port, the probing shell bottom end is provided with third connectivity mouth, is provided in the third connectivity mouth for driving object lens
The zoom motor moved up and down, the zoom motor are detachably secured on the probing shell by the fixed plate, in which:
It is provided with the first optical path in the first passage formed between first port and the third connectivity mouth, described second
The second optical path is provided in the second channel formed between port and the third connectivity mouth, in which:
First optical path successively includes collimation lens, the microcomputer between first port and the third connectivity mouth
Electric scanning galvanometer, the first lens, the second lens, dichroscope and the object lens, wherein first optical path is for conducting institute
The received laser signal of the first port is stated from first port to the third connectivity mouth；
Second optical path successively includes the object lens and the institute between the third connectivity mouth and second port
Dichroscope is stated, wherein second optical path is for conducting the collected optical signal of the object lens from the third connectivity mouth to described
Second port.
The third aspect, the embodiment of the present invention provide a kind of absorption type three dimensional non-linear laser scanning microscope, comprising:
Phosphor collection device, air extractor, scanning collection controller, femtosecond pulse laser, fiber coupling module and
The absorption type microscope detection device that second aspect of the embodiment of the present invention provides, the phosphor collection device and the fiber coupling
Module is connect with the absorption type microscope detection device fiber optic communication, and the phosphor collection device and the absorption type are micro-
Mirror detection device is electrically connected with the scanning collection controller, the air extractor and the absorption type microscope detection device
Electrical connection, in which:
The femtosecond pulse laser, for exporting pulsed laser signal to the fiber coupling module；
The fiber coupling module, for coupling the pulsed laser signal of the femtosecond pulse laser output, and
Transmit described the first of the probe of mini microscope described in the pulsed laser signal to the absorption type microscope detection device
Port；
The absorption type microscope detection device exports the pulse laser after receiving the pulsed laser signal
The signal autofluorescence substance intracellular to life entity, and obtain the fluorescence signal generated after the autofluorescence substance excitation
And second harmonic signal, and the fluorescence signal and the second harmonic signal are exported to the phosphor collection device；
The phosphor collection device converts institute after receiving the fluorescence signal and the second harmonic signal respectively
It states fluorescence signal and the second harmonic signal is corresponding electric signal；
The scanning collection controller sweeps the pulsed laser signal for controlling the mini microscope probe
Retouch and synchronous acquisition described in electric signal；
The air extractor is evacuated for the outer adsorption space to the absorption type microscope detection device,
With the negative pressure formed in the outer adsorption space.
It is provided in an embodiment of the present invention to be used to be arranged positioning formula adsorbent equipment, the absorption type microscope that mini microscope is popped one's head in
Detection device and absorption type three dimensional non-linear laser scanning microscope are removable using the magnetic effect between outer housing and pedestal
Unload and link together, sucker and coverslip formation enable to adsorbent equipment be adsorbed on the outer adsorption space on life entity skin and
For placing the built-in space of mini microscope probe, wherein it is in sealing contact to realize that coverslip is fixed on the seal of sucker,
Above-mentioned outer adsorption space is formed by the inner space of sucker；Telecontrol equipment is placed in built-in space, and is integrally attached to shell
On the side wall of body, by the first drive block and the second drive block, realize mini microscope probe in the horizontal plane along XY axis direction
Mobile, single unit system micromation after device is adsorbed on human skin, in the case where not moving adsorbent equipment, is seen to find
It examines a little, realizes the accurate positioning of mini microscope probe, obtain the imaging of different zones, so that preferably observation is wanted in observation
Imaging region, it is easy to operate, easy to use, while multiple sites can successively be imaged, finally by image by the mobile position XY
Splicing obtains bigger visual field.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is this hair
Bright some embodiments for those of ordinary skill in the art without creative efforts, can be with root
Other attached drawings are obtained according to these attached drawings.
Fig. 1 is provided in an embodiment of the present invention for the positioning formula adsorbent equipment structural representation of mini microscope probe to be arranged
Figure；
Fig. 2 is the positioning formula adsorbent equipment structural schematic diagram after combination provided in an embodiment of the present invention；
Fig. 3 is that positioning formula adsorbent equipment provided in an embodiment of the present invention passes through cross laser progress long distance positioning signal
Figure；
Fig. 4 is the sucker structure schematic diagram being arranged on pedestal in positioning formula adsorbent equipment provided in an embodiment of the present invention；
Fig. 5 is the positioning formula adsorbent equipment the schematic diagram of the section structure one after combination provided in an embodiment of the present invention；
Fig. 6 is the positioning formula adsorbent equipment the schematic diagram of the section structure two after combination provided in an embodiment of the present invention；
Fig. 7 is the structural schematic diagram of mini microscope provided in an embodiment of the present invention probe；
Fig. 8 is the first light channel structure schematic diagram in mini microscope provided in an embodiment of the present invention probe；
Fig. 9 is the positioning formula adsorbent equipment the schematic diagram of the section structure three after combination provided in an embodiment of the present invention；
Figure 10 is absorption type three dimensional non-linear laser scanning microscope structural schematic diagram provided in an embodiment of the present invention；
Figure 11 is phosphor collection apparatus structure schematic diagram provided in an embodiment of the present invention；
Figure 12 is that absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention detects human face skin
Skin tissue schematic diagram；
Figure 13 is that absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention detects human chest skin
Skin tissue schematic diagram；
Figure 14 is that the multiple detection devices of absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention are same
When detect human skin tissue schematic diagram；
Figure 15 is absorption type three dimensional non-linear laser scanning microscope detecting animal skin group provided in an embodiment of the present invention
Knit schematic diagram；
Figure 16 shows for the box composite structure that the embodiment of the present invention provides absorption type three dimensional non-linear laser scanning microscope
It is intended to；
Figure 17 is the box composite structure of absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention
Joint sealing structural schematic diagram.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
Every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
It is existing to be based on fluorescence signal and second harmonic signal, it is main to obtain the correlation detection equipment of biological cell form
If three dimensional non-linear laser scanning microscope, the form of current above-mentioned laser scanning microscope is predominantly moved by mechanical arm
The detection device installation of the i.e. laser scanning microscope of laser scanning microscope of dynamic microscope detection device on the robotic arm, is led to
The adjustment for crossing mechanical arm carrys out movement detector, is then aligned with the different institutional framework of detection human body.But above-mentioned three dimensional non-linear
The detection device based on mechanical arm in laser scanning microscope, since its volume is larger, the corresponding biggish skin of human body of popping one's head in
Area needs multiple adjustment machine so that can not be accurately positioned in concrete operations for the specific position of skin detection
Tool arm, multiple movement detector, locating effect is poor, is easy to produce offset deviation, to influence image quality.
For the ease of carrying out accurate positioning imaging to life body tissue, its cellular prion protein information is obtained, the present invention
Embodiment provides a kind of positioning formula adsorbent equipment popped one's head in for mini microscope to be arranged, and Fig. 1 provides for the embodiment of the present invention
For be arranged mini microscope probe positioning formula adsorbent equipment structural schematic diagram, as shown in Figure 1, the positioning formula adsorbent equipment
Include:
Outer housing 11, pedestal 13, coverslip 12 and the movement for driving mini microscope probe 15 to move in the horizontal plane
Device is provided with sucker 131 on pedestal 13, and sucker 131 is embedded in the sucker hole that 11 bottom of outer housing opens up, outer housing 11 and bottom
Seat 13 is detachably connected by magnetic field force, in which:
Coverslip 12 is fixed on the seal 1311 of sucker 131, and built-in space and the outer absorption for forming adsorption device are empty
Between；
Telecontrol equipment is set in built-in space, and telecontrol equipment includes fixed bracket 142, the first drive block 141 and second
Drive block 143, the first drive block 141 and the second drive block 143 are fixedly connected with fixed bracket 142 respectively, the second drive block 143
It is fixed on the side wall of outer housing 11, in which:
Second drive block 143 is for driving the first drive block 141 and fixed bracket 142 to move along the y axis together, and first
Drive block 141 is used to that mini microscope probe 15 to be driven to move along the x axis, wherein the positive cover alignments of mini microscope probe 15
Slide 12.
Specifically, provided in an embodiment of the present invention for being arranged in the positioning formula adsorbent equipment of mini microscope probe 15
Outer housing 11 and pedestal 13 be can the material of mutually magnetic constitute, or the magnetic bodies of mutual magnetic is built-in with, so that each other
It can be detachably connected by magnetic field force, and be provided with sucker 131 on pedestal 13,11 bottom of outer housing is provided with for embedding
Enter the sucker hole of sucker 131, sucker 131 is equipped with the seal 1311 being connected to space in sucker 131, when coverslip 12 covers
When on seal 1311, built-in space is formed in the positioning formula adsorbent equipment and adsorbent equipment is enabled to be adsorbed on life entity
Outer adsorption space on skin.
Wherein, the telecontrol equipment for driving mini microscope probe 15 to move in the horizontal plane is arranged in built-in space
Interior, which includes fixed bracket 142, the first drive block 141 and the second drive block 143, the first drive block 141 and second
Drive block 143 includes sliding block and fixed block, and the first fixed block of the first drive block 141 is fixed on fixed bracket 142, and first
First sliding block of drive block 141 is under the driving of first motor, sliding opposite with the first fixed block；The of second drive block 143
Two sliding blocks are fixed on fixed bracket 142, and the second fixed block of the second drive block 143 is fixed on the side wall of outer housing 11, the
Second sliding block of two drive blocks 143 is under the driving of the second motor, sliding opposite with the second fixed block；Mini microscope probe 15
It can be fixed on the first sliding block of the first drive block 141, so that the first drive block 141 is able to drive mini microscope probe
15 move along the x axis, and the second drive block 143 is able to drive the first drive block 141 and fixed bracket 142 and is fixed on first
Mini microscope probe 15 on drive block 141 moves along the y axis together, to realize adsorbent equipment in coarse positioning to life
After on body skin histology to be measured, mini microscope probe 15 can be driven dynamic along X-axis and Y direction micro-shifting, come by telecontrol equipment
It realizes in the horizontal plane to the precise positioning of mini microscope probe 15, while being realized by motor driven along X-axis and Y direction
It is mobile, so that realizing the imaging of different zones while the microscope adsorbent equipment not moved, can preferably observe desired
The imaging region of observation.The identical distance of displacement platform stepping can be made, and the image of imaging is carried out by process control simultaneously
Splicing, can be obtained the imaging results in the N times of visual field.
And after fixed bracket 142 and mini microscope probe 15 fix, the positive cover alignment of mini microscope probe 15
Slide 12, to export internal signal through coverslip 12 and receive external signal, to realize microscope zoom, it is three-dimensional at
Picture.And outer housing 11, pedestal 13, coverslip 12 and the telecontrol equipment in above-mentioned Fig. 1 are combined into one generally for being arranged
The positioning formula adsorbent equipment of mini microscope probe 15.Fig. 2 is that the positioning formula after combination provided in an embodiment of the present invention adsorbs dress
Set structural schematic diagram, as shown in Fig. 2, become on the fixed adsorbent equipments after combining of mini microscope probe 15 be provided with it is micro-
The positioning formula adsorbent equipment of type microscope probe 15, wherein mini microscope probe 15 is located in built-in space, is fixed on shell
On the side wall of body 11, and the object lens forward direction of mini microscope probe 15 is directed at coverslip, absorption type microscope detection device it is outer
Shell 11 is detachably connected with pedestal 13 by magnetic field force.
Positioning formula adsorbent equipment provided in an embodiment of the present invention for mini microscope probe to be arranged using outer housing and
Magnetic effect between pedestal is detachably connected, and sucker and coverslip formation enable to adsorbent equipment to be adsorbed on life
Outer adsorption space on body skin and the built-in space for placing mini microscope probe, wherein coverslip is fixed on sucker
Seal it is in sealing contact to realize, above-mentioned outer adsorption space is formed by the inner space of sucker；Telecontrol equipment is placed in built-in
It in space, and is integrally attached on the side wall of outer housing, by the first drive block and the second drive block, realizes that mini microscope is visited
Head is moved along XY axis direction in the horizontal plane, and single unit system micromation after device is adsorbed on human skin, is not moving absorption
While device, to find point of observation, realizes the accurate positioning of mini microscope probe, obtain the imaging of different zones, thus
Preferably the imaging region of observation is wanted in observation, easy to operate, easy to use.
It is provided in an embodiment of the present invention for the positioning of mini microscope probe to be arranged on the basis of the various embodiments described above
Formula adsorbent equipment, as shown in Figure 1, further includes:
One one word laser 16 and the 2nd 1 word laser 17, the one one word laser 16 and the 2nd 1 word laser 17
It is all set in 11 bottom of outer housing, by emitting a word laser in the transmission outside adsorption space of coverslip, in which:
One one word laser 16, for emitting the word laser being overlapped with X-axis, the 2nd 1 word laser 17, for sending out
Penetrate the word laser being overlapped with Y-axis.It is provided in an embodiment of the present invention for mini microscope to be arranged in order to facilitate coordinate detection
11 bottom of outer housing of the positioning formula adsorbent equipment of probe is also provided with two word lasers, is located at the X-axis and Y of sucker
Axis direction, by emitting a word laser, a word of the one one word laser 16 transmitting in the transmission outside adsorption space of coverslip
Laser is overlapped with X-axis, and a word laser of the 2nd 1 word laser 17 transmitting is overlapped with Y-axis, so that obtained right-angled intersection
The objective lens optical axis center that the cross anchor point of laser and the origin of XY axis and mini microscope are popped one's head in is overlapped, and cross hairs also with
XY overlapping of axles.Fig. 3 is that positioning formula adsorbent equipment provided in an embodiment of the present invention passes through cross laser progress long distance positioning signal
Figure, as shown in figure 3, when skin histology of the positioning formula adsorbent equipment 1 apart from life entity to be measured with mini microscope probe compared with
When remote, long distance positioning can be carried out by transmiting into the cross laser in outer adsorption space, then pass through telecontrol equipment again, into
The accurate positioning of row XY axis direction, thus more convenient, the accurate image-forming information for obtaining different zones.
It is provided in an embodiment of the present invention for the positioning of mini microscope probe to be arranged on the basis of the various embodiments described above
Formula adsorbent equipment, as shown in Figure 1, further includes:
Micro-camera 18 and micro lighting light source 19, micro-camera 18 and micro lighting light source 19 are all set in outer housing 11
Bottom, in which:
The micro-camera is miniature CCD or miniature CMOS；
The micro lighting light source is miniature LED or illumination fiber optic bundle；
The camera lens of micro-camera 18 is towards outer adsorption space, and the angle lens of micro-camera 18 are adjustable；
Micro lighting light source 19, for emitting optical signal in outside adsorption space.It is accurately positioned in order to further facilitate,
Outer housing 11 bottom provided in an embodiment of the present invention for the positioning formula adsorbent equipment of mini microscope probe 15 to be arranged also is set
It has set micro-camera 18 and micro lighting light source 19, has used undistorted wide-angle lens before micro-camera 18, camera lens is towards outer absorption
Space can get the biggish visual field through coverslip, when adsorbent equipment adsorbs on the skin, can be seen by micro-camera 18
The object lens of mini microscope probe 15 and the relative position of skin are examined, then micro- along XY axis direction mobile micro by telecontrol equipment
Mirror probe 15 is to the position of required observation.And the angle lens of micro-camera 18 can carry out up and down adjustment as needed；Miniature photograph
Mingguang City source 19 can emit optical signal in outside adsorption space, be used for when adsorbent equipment adsorbs on the skin, to micro-camera 18
Illumination is provided, wherein the wavelength for providing illuminating micro lighting light source is separated with the wavelength of fluorescence imaging, such as
The second harmonic signal wavelength of imaging is 390nm and the two-photon auto flourescence signals for imaging are 450-600nm, then LED
Lamp wavelength is chosen as 370nm, 635nm or infrared 850nm, 940nm.
It is provided in an embodiment of the present invention for the positioning of mini microscope probe to be arranged on the basis of the various embodiments described above
Sucker in formula adsorbent equipment further includes adsorption orifice and bleeding point, and Fig. 4 is positioning formula adsorbent equipment provided in an embodiment of the present invention
The sucker structure schematic diagram being arranged on middle pedestal, as shown in figure 4, adsorption orifice 1312 is connected with seal 1311, to pass through
Outer adsorption space is adsorbed on life entity to be measured；
Bleeding point 1313 is connected with outer adsorption space, to extract the gas in outer adsorption space.I.e. the present invention is implemented
The sucker 131 being set on pedestal 13 in the absorption type microscope detection device that example provides further includes absorption in addition to seal 1311
Mouth 1312 and bleeding point 1313, and seal 1311 is connected with adsorption orifice 1312, forms the inner space of sucker 131, works as lid
When slide sealing is fixed on seal 1311, the inner space of sucker 131 just forms outer adsorption space, and adsorption orifice 1312 is logical
The outer adsorption space is crossed to be adsorbed on life entity to be measured；Above-mentioned bleeding point 1313 is connected to outer adsorption space, passes through the bleeding point
1313, the gas in outer adsorption space can be extracted, to form the negative pressure in outer adsorption space.In the effect of external pressure
Under, absorption type microscope detection device is adsorbed on the tissues such as the skin of life entity to be measured.
Wherein, Fig. 5 is the positioning formula adsorbent equipment the schematic diagram of the section structure one after combination provided in an embodiment of the present invention, such as
Shown in Fig. 5, sucker is embedded in the sucker hole of outer housing 11, and coverslip 12 is fixed on sucker, and mini microscope probe 15 passes through
Telecontrol equipment 14 is fixed on the side wall of outer housing 11, and the object lens forward direction of mini microscope probe 15 is directed at coverslip 12.And
Mini microscope probe 15 is located in built-in space 1314, and absorption type microscope detection device passes through shape in outer adsorption space 1315
At negative pressure, be adsorbed on the tissues such as the skin of life entity to be measured, the bleeding point 1313 of sucker on outer housing for placing
The through-hole 1111 of exhaust pipe is connected, and the one one word laser 16 and micro-camera 18 are fixed on the bottom of outer housing 11.
Wherein, Fig. 6 is the positioning formula adsorbent equipment the schematic diagram of the section structure two after combination provided in an embodiment of the present invention, such as
Shown in Fig. 6, mini microscope probe 15 is located in built-in space, and the object lens forward direction cover alignment glass of mini microscope probe 15
Piece, the 2nd 1 word laser 17 and micro lighting light source 19 are fixed in the bottom of outer housing 11.
It is provided in an embodiment of the present invention for the positioning of mini microscope probe to be arranged on the basis of the various embodiments described above
Fixation bracket in formula adsorbent equipment includes X-axis bracket and Y-axis bracket, and X-axis bracket is mutually perpendicular to be fixedly connected with Y-axis bracket,
Wherein:
X-axis bracket with the first drive block is detachable is fixedly connected, Y-axis bracket and the second drive block it is detachable it is fixed even
It connects.Fixation bracket in positioning formula adsorbent equipment i.e. provided in an embodiment of the present invention for mini microscope probe to be arranged is L
Type comprising X-axis bracket and Y-axis bracket, X-axis bracket are mutually perpendicular to be fixedly connected with Y-axis bracket, X-axis bracket and the first driving
First fixed frame of block is detachably fixed connection, and Y-axis bracket is detachably fixed with the second sliding block of the second drive block and connect, thus
Realize that mini microscope probe under the driving of telecontrol equipment, can be moved along XY axis direction.
The embodiment of the present invention also provides a kind of absorption type microscope detection device, the absorption type microscope detection device packet
It includes:
The positioning formula for mini microscope probe to be arranged that mini microscope probe and the various embodiments described above provide is inhaled
Adsorption device, mini microscope probe are fixed on the first drive block, in which:
Mini microscope probe includes probing shell and fixed plate, and probing shell top is provided with the first port and second
Port, probing shell bottom end are provided with third connectivity mouth, and the zoom electricity for driving object lens to move up and down is provided in third connectivity mouth
Machine, zoom motor are detachably secured on probing shell by fixed plate, in which:
The first optical path is provided in the first passage formed between first port and third connectivity mouth, the second port and third are logical
The second optical path is provided in the second channel formed between mouthful, in which:
First optical path successively include collimation lens between the first port and third connectivity mouth, micro electromechanical scanning galvanometer,
First lens, the second lens, dichroscope and object lens, wherein the first optical path is for conducting the received laser signal of the first port
From the first port to third connectivity mouth；
Second optical path successively includes the object lens and dichroscope between third connectivity mouth and the second port, wherein the second light
Road is for conducting the collected optical signal of object lens from third connectivity mouth to the second port.
Specifically, Fig. 7 is the structural schematic diagram of mini microscope provided in an embodiment of the present invention probe, as shown in fig. 7, on
Port there are three opening up on the probing shell 151 of mini microscope probe is stated, two passes are formd between three ports, two
Two optical paths, respectively the first optical path and the second optical path, this two passes formed between three ports are provided in channel
Between have the part of one section of coincidence, above-mentioned two optical paths shared dichroscope 1544 and object are set in the passage portion of coincidence
Mirror 1545, wherein the first optical path is transmitting optical path, successively includes the collimation between the first port 154 and third connectivity mouth 156
Lens 157, micro electromechanical scanning galvanometer 1541, the first lens 1542, the second lens 1543, dichroscope 1544 and object lens
1545, i.e. laser signal enters from the first port 154, is conducted by the first optical path, projects from third connectivity mouth 156, and be incident on
The directional light of first port is projected in a manner of directional light after two lens；Second optical path is to collect optical path, and being used for will
Collected optical signal, from the output of the second port 155 to the acquisition device with mini microscope probe connection；First optical path and
The shared object lens of two optical paths are mounted in high-precision zoom motor 153, under the drive of zoom motor, can be moved up and down, wherein
Zoom motor 153 is detachably secured on probing shell 151 by fixed plate 152；And first the laser signal of optic path exist
, can be in vertical forward entrance to object lens 1545 under the reflection of dichroscope 1544, the laser of the dichroscope the first optical path of reflection
Signal, the optical signal of transmission the second optical path acquisition, and the pitch angle of the dichroscope 1544 is adjustable, object lens 1545 are infinite
Remote object lens, wherein the stepping accuracy of above-mentioned zoom motor is less than 1 micron.
Wherein, Fig. 8 is the first light channel structure schematic diagram in mini microscope provided in an embodiment of the present invention probe, such as Fig. 8
It is shown, the of the second focus 73 of the first lens 71 and the second lens 72 in mini microscope provided in an embodiment of the present invention probe
One focus is overlapped, and micro electromechanical scanning galvanometer is located in the first focus 75 of the first lens 71, and the rear pupil face 74 of object lens is located at second
In second focus of lens 72, in the light beam and output being incident on micro electromechanical scanning galvanometer to the light after object lens on pupil face 74
Conjugation is formed between beam.In the first light channel structure in mini microscope probe i.e. provided in an embodiment of the present invention, for scanning
The micro electromechanical scanning galvanometer of incident laser signal is installed on the first focal point 75 of the first lens 71, and the second of the first lens 71 is burnt
First focus of point 73 and the second lens 72 is overlapped, by adjusting zoom motor, so that the rear pupil face 74 of object lens is located at second thoroughly
Second focal point of mirror 72, hereby, it is achieved that the light beam being incident on micro electromechanical scanning galvanometer and output are to after object lens on pupil face 74
Light beam between formed conjugation so that D2/D1=F2/F1, wherein D2For the hot spot for being incident on the light beam in pupil face 74 after object lens
Diameter, D1For be incident on 71 first focal plane of the first lens light beam spot diameter, F2For the focal length of the second lens 72, F1It is
The focal length of one lens 71；tanθ2/tanθ1=F1/F2, θ1For scan angle of the incident beam at 71 first focal plane of the first lens, θ2
Incidence is swashed by adjusting the first lens 71 and the second lens 72 for scan angle of the incident beam after object lens at pupil face 74
Light light beam is expanded, so that being full of the rear pupil face 74 of entire object lens, Lai Shixian high-resolution imaging into the light beam of object lens.
Absorption type microscope detection device provided in an embodiment of the present invention, which is used, installs object lens additional in positioning formula adsorbent equipment
Can up and down adjustment mini microscope probe, pass through telecontrol equipment drive mini microscope pop one's head in horizontal plane it is mobile carry out it is different
Imaging region is accurately positioned, and drives mini microscope probe to move up and down by zoom motor, to life entity cellular morphology to be measured
Carry out different depth accurately detecting, be combined with each other under, thus realize to the three of life body tissue different zones different depth
Dimension is accurately positioned detection, obtains the cyto-architectural high-resolution imaging of different zones different depth, and method is simple, operation side
Just.
On the basis of the various embodiments described above, absorption type microscope detection device provided in an embodiment of the present invention, further includes:
Liquid lens, the liquid lens is between the collimation lens and the micro electromechanical scanning galvanometer, to be formed
The first new optical path, the first new optical path successively include the collimation between first port and the third connectivity mouth
Lens, the liquid lens, the micro electromechanical scanning galvanometer, first lens, second lens, the dichroscope with
And the object lens.First in mini microscope probe in absorption type microscope detection device i.e. provided in an embodiment of the present invention
Optical path can increase liquid lens 158, and Fig. 9 is that the positioning formula adsorbent equipment cross-section structure after combination provided in an embodiment of the present invention shows
It is intended to three, as shown in figure 9, realizing by the upper increase liquid lens 158 of mini microscope 15 structures of probe shown in Fig. 9
Zoom function.The position of liquid lens 158 is shown in Fig. 9 between collimation lens 157 and micro electromechanical scanning galvanometer 1541
Laser signal is being directional light after collimation lens 157 in original optical path structure.After increasing liquid lens 158, by the liquid
Body lens 158, which apply voltage or current, makes 158 surface of liquid lens generate the bending accordingly arrived, and then generates not to directional light
Same focal power.Specific optical path are as follows: laser signal is incident on liquid lens 158 from fiber exit in parallel after collimation lens,
Corresponding focal power is generated from liquid lens 158 according to the voltage or current signal of load, outgoing is converging or diverging with light process
It is converged on sample after micro electromechanical scanning galvanometer 1541, the first lens, the second lens, dichroscope and object lens.Wherein, liquid
The focus for the laser signal that the power variation that lens 158 introduce can be such that object lens mouth is emitted is moved forward and backward in longitudinal direction, and
The response speed of liquid lens 158 is very fast, and quick longitudinal direction may be implemented in KHz magnitude in scan frequency
Scanning imagery.Wherein, liquid lens 158 is equivalent to parallel plate glass when not applying voltage or current signal, believes laser
Number without focal power and the focus after object lens will not be made to generate any offset.When specifically used, the liquid lens 158 and zoom
Motor is complementary, adjusts object lens position by zoom motor, after coarse adjustment to corresponding depth position, system is switched to liquid lens
158 zoom scan modes carry out quick three-dimensional imaging to sample, wherein when absorption type microscope detection device is not installing change
When burnt motor, zoom adjustment can also be carried out only by liquid lens.
It is provided in an embodiment of the present invention for the positioning of mini microscope probe to be arranged on the basis of the various embodiments described above
Outer housing in formula adsorbent equipment includes first shell and second shell, as shown in Figure 1, in which:
Accommodation space is provided in first shell 111, telecontrol equipment is set in accommodation space, first shell 111 and
Two shells 112 are detachable to be fixedly connected.The positioning formula i.e. provided in an embodiment of the present invention popped one's head in for mini microscope to be arranged
Outer housing in adsorbent equipment includes two parts, and respectively first shell 111 and second shell 112, first shell 111 are used for
The accommodation space of telecontrol equipment is set, i.e., the sucker hole in above-described embodiment is also disposed at the bottom of first shell 111, and sucker is embedding
Enter in the sucker hole of first shell 111, in conjunction with coverslip and second shell 112, the outer suction in above-described embodiment can be formed
Attached space and built-in space, wherein first shell 111 and second shell 112 can be detachably fixed connection by screw, so side
Just the assembling, disassembly of whole device and the replacement of component.
The embodiment of the present invention also provides a kind of absorption type three dimensional non-linear laser scanning microscope, and Figure 10 is that the present invention is implemented
The absorption type three dimensional non-linear laser scanning microscope structural schematic diagram that example provides, as shown in Figure 10, the laser scanning microscope
Include:
Phosphor collection device 56, air extractor 52, scanning collection controller 531, femtosecond pulse laser 541, optical fiber coupling
Mold block 542 and absorption type microscope detection device 51 provided by the above embodiment, phosphor collection device 56 and fiber coupling
Module 542 is connect with 51 fiber optic communication of absorption type microscope detection device, and phosphor collection device 56 and absorption type microscope are visited
Device 51 to be surveyed to be electrically connected with scanning collection controller 531, air extractor 52 is electrically connected with absorption type microscope detection device 51,
Wherein:
Femtosecond pulse laser 541, for exporting pulsed laser signal to fiber coupling module 542；
Fiber coupling module 542 for coupling the pulsed laser signal of the output of femtosecond pulse laser 541, and transmits arteries and veins
Impulse optical signal the first port that mini microscope is popped one's head in into absorption type microscope detection device；
Absorption type microscope detection device, after receiving pulsed laser signal, output pulsed laser signal to life entity
Intracellular autofluorescence substance, and the fluorescence signal that generates and secondary humorous is obtained after the excitation of autofluorescence substance by object lens
Wave signal, and fluorescence signal and second harmonic signal are exported to phosphor collection device 56；
Phosphor collection device 56, after receiving fluorescence signal and second harmonic signal, difference conversion fluorescence signal and two
Rd harmonic signal is corresponding electric signal；
Scanning collection controller 531 is scanned pulsed laser signal for controlling mini microscope probe, Yi Jitong
Step acquisition electric signal；
Air extractor 52 is evacuated for the outer adsorption space to absorption type microscope detection device, to form outer inhale
Negative pressure in attached space.
Specifically, absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention includes phosphor collection dress
Set 56, air extractor 52, scanning collection controller 531, femtosecond pulse laser 541, fiber coupling module 542 and absorption type
Microscope detection device 51, so that formation can be adsorbed on human skin or go deep into the three dimensional non-linear that human body stomach is detected
Laser scanning microscope, wherein femtosecond pulse laser 541 can be used for exciting human Skin Cell with emission pulse laser signal
In autofluorescence substance, generate multiphoton fluorescence signal and second harmonic signal, the femtosecond pulse including using 920nm
FAD and collagen in 541 activated cell of device, excite the fluorescence signal of 500-600nm and the second harmonic signal of 460nm,
And the autofluorescences substance such as FAD in 541 activated cell of femtosecond pulse laser for passing through 780nm or NADH, to generate phase
The fluorescence signal and second harmonic signal answered；
Wherein, phosphor collection device 56 is integrated with two paths of signals and collects optical path, respectively fluorescence signal collection optical path and two
Rd harmonic signal collects optical path, the collection respectively of Lai Shixian fluorescence signal and second harmonic signal；Scanning collection controller 531 is controlled
Scanning galvanometer in mini microscope probe processed is scanned pulsed laser signal and autofluorescence substance is excited to generate fluorescence
Signal and second harmonic signal, and acquire 56 conversion fluorescence signal of phosphor collection device and second harmonic signal obtain first
Electric signal and the second electric signal；Air extractor 52 includes mainly aspiration pump, is connected with exhaust pipe, exhaust pipe and above-mentioned implementation
Bleeding point in example is connected, and extraction valve is arranged in exhaust pipe, extraction valve is electrically connected with air extractor 52, and air extractor 52 passes through
The switch of extraction valve and the size of opening and closing are adjusted, the extraction flow of exhaust pipe is controlled, to realize external adsorption space
Pumping control, and then the negative pressure in outer adsorption space is adjusted, so that adsorbent equipment is adsorbed on life entity by the effect of atmospheric pressure
The tissues such as skin, stomach, and the absorption type three dimensional non-linear laser scanning microscope may include that two-photon scans according to classification
Microscope and multi-photon flying-spot microscope etc., wherein when femtosecond pulse laser can replaced by common continuous light laser
In the case where generation, increase aperture, it is burnt micro- for copolymerization also to can adjust the absorption type three dimensional non-linear laser scanning microscope
Mirror.Wherein, the resolution ratio of the absorption type three dimensional non-linear laser scanning microscope may be configured as 800nm, and visual field can be 200
* 200 microns of micron, image taking speed can be 26 frames (256*256 pixel) or 13 frames (512*512 pixel).
Absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention is using phosphor collection device, pumping
Device, scanning collection controller, femtosecond pulse laser, fiber coupling module and absorption type microscope detection device, thus
Formation can be adsorbed on human skin or go deep into the three dimensional non-linear laser scanning microscope that human body stomach is detected, and pass through tune
Whole mini microscope probe is adjusted focal length at a distance from coverslip, realizes the 3-D scanning of laser scanning microscope, passes through
Autofluorescence substance obtains multiphoton fluorescence signal and second harmonic signal in femtosecond pulse laser activated cell, realizes laser
Flying-spot microscope is non-linear, collects fluorescence signal and second harmonic signal by phosphor collection device, and be converted to corresponding
Electric signal, and then the corresponding fluorescent image etc. for reflecting cell tissue structure is obtained by the electric signal, wherein absorption type is micro-
The use of mirror detection device can pop one's head on mini microscope to avoid life entity activity and generate vibration influence, to avoid vibration shadow
Ring arrive image quality, and above-mentioned laser scanning microscope can realize a variety of imaging patterns include XY imaging, XZ imaging and 3D at
Picture, wherein XY is imaged as carrying out transversal scanning imaging in a certain level of eucaryotic cell structure certain depth, and XZ is imaged as past from surface layer
The XZ cross-sectional imaging of lower certain depth, 3D are imaged as carrying out XY imaging from surface layer certain depth down, each depth, reconstruct
At 3D rendering, equipment operation is simple, easy to use.
On the basis of the various embodiments described above, Figure 11 is phosphor collection apparatus structure provided in an embodiment of the present invention signal
Figure, as shown in figure 11, phosphor collection device provided in an embodiment of the present invention includes fiber optic universal interface 881, the first photomultiplier transit
Pipe 882, the second photomultiplier tube 883 and the first receipts between fiber optic universal interface 881 and the first photomultiplier tube 882
Collect optical path, the second collection optical path between fiber optic universal interface 881 and the second photomultiplier tube 883, in which:
First collection optical path successively includes coupling collecting lens 81, infrared fileter 82, the filter of the first dichroscope 83, first
Mating plate 84 and the first collecting lens 85, wherein the first collection optical path is used to collect the fluorescence letter that phosphor collection device receives
Number, the first photomultiplier tube 882 is the first electric signal for conversion fluorescence signal；
Second collection optical path successively includes coupling collecting lens 81, infrared fileter 82, the first dichroscope the 83, the 2nd 2
To Look mirror 86, the second optical filter 87 and the second collecting lens 88, wherein the second collection optical path is for collecting phosphor collection device
The second harmonic signal received, the second photomultiplier tube 883 is for converting second harmonic signal as the second electric signal.That is this hair
The phosphor collection device that bright embodiment provides has simple two-way signal collecting function, is integrated with two-way optical path, wherein first collects light
The first dichroscope 83 in road is transmission fluorescence signal, the dichroscope of reflected second harmonics, the second dichroscope 86 and the
One dichroscope 83 is same dichroscope, is used for reflected second harmonics, and the first optical filter 84 is filtered for transmiting fluorescence signal
Except remaining interference signal, the second optical filter 87 filters out remaining interference signal for transmiting corresponding second harmonic signal, for example,
When using the intracellular autofluorescence substance of 880nm femto second optical fiber laser exciting human surface skin, the two of 390nm can be obtained
The two-photon auto flourescence signals of rd harmonic signal and 450-600nm, are passed through by 420nm above wavelength, and 420 or less wavelength are anti-
The dichroscope penetrated i.e. the first dichroscope 83 can separate two-way fluorescence, use the first optical filter 84 of 390 ± 20nm respectively
With the second optical filter 87 available clean second harmonic signal and fluorescence signal of 450-600nm.
On the basis of the various embodiments described above, absorption type three dimensional non-linear scan laser microphotograph provided in an embodiment of the present invention
Mirror further includes industrial personal computer, and as shown in Figure 10, industrial personal computer 532 is electrically connected with scanning collection controller 531, in which:
Industrial personal computer 532 is for obtaining collected first electric signal of scanning collection controller 531 and the second electric signal, and base
The first fluorescent image is generated in the first electric signal and the second fluorescent image is generated based on the second electric signal.That is the embodiment of the present invention
The absorption type three dimensional non-linear laser scanning microscope of offer further includes the industrial personal computer being electrically connected with scanning collection controller 531
532, which is based on the first electric signal and generates the first fluorescent image and generate the second fluorogram based on the second electric signal
Picture can be respectively used to display eucaryotic cell structure and fibre structure information, control software is wherein equipped on industrial personal computer, soft by controlling
Part sends control instruction to scanner, to control scanning collection controller, to obtain above-mentioned first electric signal and the second telecommunications
Number.
On the basis of the various embodiments described above, absorption type three dimensional non-linear scan laser microphotograph provided in an embodiment of the present invention
Mirror further includes display, and as shown in Figure 10, display 55 is electrically connected with industrial personal computer 532, for showing the first fluorescent image and the
Two fluorescent images.Absorption type three dimensional non-linear laser scanning microscope i.e. provided in an embodiment of the present invention further includes for showing the
The display 55 of one fluorescent image and the second fluorescent image, by display 55, staff can directly acquire the first fluorescence
The relevant information of image and the second fluorescent image.
On the basis of the various embodiments described above, absorption type three dimensional non-linear scan laser microphotograph provided in an embodiment of the present invention
Absorption type microscope detection device in mirror is multiple.Phosphor collection device and fiber coupling mould i.e. provided in an embodiment of the present invention
Block can be connect with multiple absorption type microscope detection device fiber optic communications simultaneously, i.e., sweep in an absorption type three dimensional non-linear laser
It retouches and integrates multiple detection devices in microscopic system, to be detected while realization to life entity different tissues position, to carry out
Comparative analysis.
On the basis of the various embodiments described above, absorption type three dimensional non-linear scan laser microphotograph provided in an embodiment of the present invention
Mirror further includes adjusting optical fiber, for phosphor collection device and fiber coupling module respectively with absorption type microscope detection device it
Between optical fiber transmit connection, in which:
Adjust the adjustable in length of optical fiber.Absorption type three dimensional non-linear scan laser microphotograph i.e. provided in an embodiment of the present invention
Phosphor collection device and fiber coupling module in mirror pass through length-adjustable adjusting optical fiber respectively and absorption type microscope detects
Device carries out optical fiber transmission connection and carries out flexible movement detector to realize according to different experiments scene needs, avoid limited
The limitation of fiber lengths, wherein the adjustable in length of optical fiber is adjusted, to realize various fields by the optical fiber for replacing different length
The application of conjunction can carry out the optical fiber replacement of different length at any time as needed.
In order to more clearly illustrate absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention
Application scenarios are now described further with legend, and Figure 12 is that absorption type three dimensional non-linear laser provided in an embodiment of the present invention is swept
Retouch microscope detection human facial skin's schematic illustration of tissue, by the air extracting function of air extractor 52, will adsorb as shown in 12
Formula microscope detection device 51 is adsorbed on human face, wherein scanning collection controller and industry control are integrated in first device 53
Machine, industrial personal computer are electrically connected with display 55, and second device 54 is integrated with femtosecond pulse laser, fiber coupling module and fluorescence and receives
Acquisition means, fiber coupling module and phosphor collection device are transmitted with 51 optical fiber of absorption type microscope detection device and are connected, wherein
The absorption type three dimensional non-linear laser scanning microscope working principle is identical as the various embodiments described above, and details are not described herein again.
Wherein, Figure 13 is that absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention detects human body chest
Portion's skin histology schematic diagram, as shown in 13, by the air extracting function of air extractor 52, by absorption type microscope detection device 51
It is adsorbed on human chest, wherein scanning collection controller and industrial personal computer, industrial personal computer and display 55 are integrated in first device 53
Electrical connection, second device 54 are integrated with femtosecond pulse laser, fiber coupling module and phosphor collection device, fiber coupling module
It transmits and connects with 51 optical fiber of absorption type microscope detection device with phosphor collection device, wherein the absorption type three dimensional non-linear
Laser scanning microscope working principle is identical as the various embodiments described above, and details are not described herein again.
Wherein, Figure 14 is the multiple detection dresses of absorption type three dimensional non-linear laser scanning microscope provided in an embodiment of the present invention
It sets while detecting human skin tissue schematic diagram, as shown in 14, by the air extracting function of air extractor 52, by multiple absorption types
Microscope detection device 51 is adsorbed on human face, chest and leg simultaneously respectively, wherein scanning is integrated in first device 53
Acquisition controller and industrial personal computer, industrial personal computer are electrically connected with display 55, and second device 54 is integrated with femtosecond pulse laser, optical fiber
Coupling module and phosphor collection device, fiber coupling module and phosphor collection device with 51 light of absorption type microscope detection device
Fibre transmission connection, thus, it realizes under the action of multiple absorption type microscope detection devices, while detecting human body areas of skin
Skin tissue structure, it is easy to operate, easy to use, and due to using optical fiber between absorption type microscope detection device and second device
Transmission connection, fiber lengths can be adjusted, and human body to be measured is move freely.Wherein, the absorption type three-dimensional is non-thread
Property laser scanning microscope working principle is identical as the various embodiments described above, and details are not described herein again.Figure 15 mentions for the embodiment of the present invention
The absorption type three dimensional non-linear laser scanning microscope detecting animal skin histology schematic diagram of confession as shown in figure 15 equally can be with
By the air extracting function of air extractor 52, absorption type microscope detection device 51 is adsorbed on the skin histology of life entity,
In, scanning collection controller and industrial personal computer are integrated in first device 53, industrial personal computer is electrically connected with display 55, second device 54
It is integrated with femtosecond pulse laser, fiber coupling module and phosphor collection device, fiber coupling module and phosphor collection device are equal
It transmits and connects with 51 optical fiber of absorption type microscope detection device, working principle is identical as the various embodiments described above.
For the absorption type three dimensional non-linear laser scanning microscope that the various embodiments described above provide, the embodiment of the present invention is also mentioned
Another specific embodiment is supplied, Figure 16 provides absorption type three dimensional non-linear laser scanning microscope for the embodiment of the present invention
Box combining structure schematic diagram, as shown in figure 16, the scanning collection control of the absorption type three dimensional non-linear laser scanning microscope
Device 531, industrial personal computer 532, air extractor 52, phosphor collection device 56 and femtosecond pulse laser processed and fiber coupling are integrated in
Integration module 540 together, is integrated in Portable suitcase together, there is the industrial personal computer with display screen in case, and suitcase
Display 55 is integrated on case lid；Absorption type microscope detection device 51 is adsorbed on the skin histology of human body to be measured, with cabinet
Interior fiber coupling is connected with 56 fiber optic communication of phosphor collection device, is connect with aspiration pump by exhaust pipe, attaching plug with
Scanning collection controller 531 and industrial personal computer 532 are electrically connected.Wherein Figure 17 is that absorption type three-dimensional provided in an embodiment of the present invention is non-thread
The joint sealing structural schematic diagram of the box composite structure of property laser scanning microscope is integrated in display on case lid as shown in figure 17
55 integrate with the cabinet for being equipped with modules, facilitate whole equipment mobile, and replacement workplace, and the display
Device 55 when in use, can be placed on cabinet in addition, to facilitate staff to obtain the information on display.When having used the suction
After attached formula three dimensional non-linear laser scanning microscope, staff can portable equipment case, convenient changing workplace is especially being cured
Institute, laboratory or outdoor location, can be more convenient using the equipment.
It should be further noted that the absorption type three dimensional non-linear laser scanning microscope that the various embodiments described above provide,
In the wavelength for changing femtosecond pulse laser and after adjusting the filter range of each optical filter, fluorescence and non-SHG (Second in part
Harmonic Generation, second harmonic generate) the active tissue of signal, CARS signal can be collected, so as to adjust for
The miniature CARS microscope of absorption type, specific adjusting parameter are configured according to specific needs.
Above-mentioned, although the foregoing specific embodiments of the present invention is described with reference to the accompanying drawings, not to invention protection scope
Limitation, those skilled in the art should understand that, based on the technical solutions of the present invention, those skilled in the art are not required to
The above is only preferred embodiment of the present application, are not intended to restrict the invention, for those skilled in the art, this
Application can have various modifications and variations.Within the spirit and principles of this application, made any modification, equivalent replacement,
Improve etc., it should be included within the scope of protection of this application.
The apparatus embodiments described above are merely exemplary, wherein unit can be as illustrated by the separation member
Or may not be and be physically separated, component shown as a unit may or may not be physical unit, i.e.,
It can be located in one place, or may be distributed over multiple network units.It can select according to the actual needs therein
Some or all of the modules achieves the purpose of the solution of this embodiment.Those of ordinary skill in the art are not paying creative labor
In the case where dynamic, it can understand and implement.
Finally, it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
Present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: it still may be used
To modify the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features；
And these are modified or replaceed, technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution spirit and
Range.
Claims (11)
1. a kind of for the positioning formula adsorbent equipment of mini microscope probe to be arranged characterized by comprising
Outer housing, pedestal, coverslip and the telecontrol equipment for driving mini microscope probe to move in the horizontal plane, the pedestal
On be provided with sucker, the sucker is embedded in the sucker hole that the outer housing bottom opens up, and the outer housing and the pedestal are logical
Magnetic field force is crossed to be detachably connected, in which:
The coverslip is fixed on the seal of the sucker, and built-in space and the outer absorption for forming the adsorption device are empty
Between；
The telecontrol equipment is set in the built-in space, and the telecontrol equipment includes fixed bracket, the first drive block and the
Two drive blocks, first drive block and second drive block are fixedly connected with the fixed bracket respectively, and described second drives
Motion block is fixed on the side wall of the outer housing, in which:
Second drive block is described for driving first drive block and the fixed bracket to move along the y axis together
First drive block is for driving mini microscope probe to move along the x axis, wherein the positive alignment of mini microscope probe
The coverslip.
2. according to claim 1 for the positioning formula adsorbent equipment of mini microscope probe to be arranged, which is characterized in that also
Include:
One one word laser and the 2nd 1 word laser, the one one word laser and the 2nd 1 word laser are all provided with
It is placed in the outer housing bottom, emits a word laser into the outer adsorption space by transmiting the coverslip, in which:
The one one word laser, for emitting the word laser being overlapped with X-axis；The 2nd 1 word laser, for sending out
Penetrate the word laser being overlapped with Y-axis.
3. according to claim 1 or 2 for the positioning formula adsorbent equipment of mini microscope probe to be arranged, feature exists
In, further includes:
Micro-camera and micro lighting light source, the micro-camera and the micro lighting light source are all set in the outer housing bottom
Portion, in which:
The micro-camera is miniature CCD or miniature CMOS；
The micro lighting light source is miniature LED or illumination fiber optic bundle；
The camera lens of the micro-camera is towards the outer adsorption space, and the angle lens of the micro-camera are adjustable；
The micro lighting light source, for emitting into the outer adsorption space for providing the optical signal of illumination.
4. according to claim 1 or 2 for the positioning formula adsorbent equipment of mini microscope probe to be arranged, feature exists
In the sucker further includes adsorption orifice and bleeding point, and the adsorption orifice is connected with the seal, to pass through the outer suction
Attached space is adsorbed on life entity to be measured；
The bleeding point is connected with the outer adsorption space, to extract the gas in the outer adsorption space.
5. according to claim 1 or 2 for the positioning formula adsorbent equipment of mini microscope probe to be arranged, feature exists
In, the fixed bracket includes X-axis bracket and Y-axis bracket, and the X-axis bracket is mutually perpendicular to be fixedly connected with the Y-axis bracket,
Wherein:
The X-axis bracket with first drive block is detachable is fixedly connected, the Y-axis bracket and second drive block can
It is detachably fixed connection.
6. a kind of absorption type microscope detection device characterized by comprising
Mini microscope probe and the described in any item positioning formulas for mini microscope probe to be arranged of claim 1-5 are inhaled
Adsorption device, the mini microscope probe are fixed on first drive block, in which:
Mini microscope probe includes probing shell and fixed plate, the probing shell top be provided with the first port and
Second port, the probing shell bottom end are provided with third connectivity mouth, are provided in the third connectivity mouth for driving object lens or more
Mobile zoom motor, the zoom motor are detachably secured on the probing shell by the fixed plate, in which:
The first optical path, second port are provided in the first passage formed between first port and the third connectivity mouth
The second optical path is provided in the second channel formed between the third connectivity mouth, in which:
First optical path successively includes collimation lens between first port and the third connectivity mouth, micro electronmechanical sweeps
Galvanometer, the first lens, the second lens, dichroscope and the object lens are retouched, wherein first optical path is for conducting described the
The received laser signal of one port is from first port to the third connectivity mouth；
Second optical path successively includes the object lens and described two between the third connectivity mouth and second port
To Look mirror, wherein second optical path is for conducting the collected optical signal of the object lens from the third connectivity mouth to described second
Port.
7. absorption type microscope detection device according to claim 6, which is characterized in that the mini microscope probe,
Further include:
Liquid lens, the liquid lens is between the collimation lens and the micro electromechanical scanning galvanometer, to be formed newly
First optical path, the first new optical path successively include that the collimation between first port and the third connectivity mouth is saturating
Mirror, the liquid lens, the micro electromechanical scanning galvanometer, first lens, second lens, the dichroscope and
The object lens.
8. a kind of absorption type three dimensional non-linear laser scanning microscope characterized by comprising
Phosphor collection device, air extractor, scanning collection controller, femtosecond pulse laser, fiber coupling module and right
It is required that absorption type microscope detection device described in 6 or 7, the phosphor collection device and the fiber coupling module are and institute
State absorption type microscope detection device fiber optic communication connection, the phosphor collection device and the absorption type microscope detection device
It is electrically connected with the scanning collection controller, the air extractor is electrically connected with the absorption type microscope detection device,
In:
The femtosecond pulse laser, for exporting pulsed laser signal to the fiber coupling module；
The fiber coupling module for coupling the pulsed laser signal of the femtosecond pulse laser output, and transmits
First port of the probe of mini microscope described in the pulsed laser signal to the absorption type microscope detection device；
The absorption type microscope detection device exports the pulsed laser signal after receiving the pulsed laser signal
The autofluorescence substance intracellular to life entity, and obtained by the object lens and to be generated after autofluorescence substance excitation
Fluorescence signal and second harmonic signal, and export the fluorescence signal and the second harmonic signal to the phosphor collection and fill
It sets；
The phosphor collection device is converted described glimmering respectively after receiving the fluorescence signal and the second harmonic signal
Optical signal and the second harmonic signal are corresponding electric signal；
The scanning collection controller is scanned the pulsed laser signal for controlling the mini microscope probe,
And electric signal described in synchronous acquisition；
The air extractor is evacuated, with shape for the outer adsorption space to the absorption type microscope detection device
At the negative pressure in the outer adsorption space.
9. absorption type three dimensional non-linear laser scanning microscope according to claim 8, which is characterized in that the fluorescence is received
Acquisition means include fiber optic universal interface, the first photomultiplier tube, the second photomultiplier tube and are located at the fiber optic universal interface
First between first photomultiplier tube collects optical path, is located at the fiber optic universal interface and second photomultiplier transit
Second between pipe collects optical path, in which:
It is described first collection optical path successively include coupling collecting lens, infrared fileter, the first dichroscope, the first optical filter with
And first collecting lens, wherein the first collection optical path for collecting the fluorescence signal that phosphor collection device receives,
First photomultiplier tube is for converting the fluorescence signal as the first electric signal；
It is described second collection optical path successively include the coupling collecting lens, the infrared fileter, first dichroscope,
Second dichroscope, the second optical filter and the second collecting lens, wherein the second collection optical path is for collecting phosphor collection
The second harmonic signal that device receives, second photomultiplier tube is for converting the second harmonic signal as second
Electric signal.
10. absorption type three dimensional non-linear laser scanning microscope according to claim 8, which is characterized in that the absorption
Formula microscope detection device is multiple.
11. absorption type three dimensional non-linear laser scanning microscope according to claim 8, which is characterized in that further include adjusting
Save optical fiber, for the phosphor collection device and the fiber coupling module respectively with the absorption type microscope detection device it
Between optical fiber transmit connection, in which:
The adjustable in length for adjusting optical fiber.
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