CN211014821U

CN211014821U - Microscope

Info

Publication number: CN211014821U
Application number: CN201921266397.3U
Authority: CN
Inventors: 不公告发明人
Original assignee: Suzhou Yibolun Photoelectric Instrument Co ltd
Current assignee: Suzhou Yibolun Photoelectric Instrument Co ltd
Priority date: 2019-08-06
Filing date: 2019-08-06
Publication date: 2020-07-14
Anticipated expiration: 2029-08-06

Abstract

The utility model relates to the technical field of optical imaging, in particular to a microscope, which comprises a light source transmitter and a collecting lens, wherein the light source transmitter is a laser transmitter, and a collimating lens, a cylindrical lens, a first scanner, a focusing lens, a wave plate, a second scanner and the collecting lens are sequentially arranged on a light path between the laser transmitter and the collecting lens; the first scanner and the second scanner are both dichroic mirror scanners, the first scanner can rotate along the vertical direction of the light path, and the second scanner can move along the direction of the light path; an objective lens is arranged behind the first scanner, and the cylindrical lens is used for focusing the laser collimated by the collimating lens into a linear focus on the surface of the first scanner. This scheme is used for solving among the prior art desk-top many photon microscope can only realize two-dimensional scanning formation of image, and structural weight is heavy, bulky, still has the technical problem that can't be applied to clinically in addition.

Description

Microscope

Technical Field

The utility model relates to an optical imaging technical field specifically is a microscope.

Background

For high resolution neuroscience research on experimental animals, multiphoton microscopy is commonly employed as a technique for noninvasive optical brain imaging. In the prior art, when a desktop multi-photon microscope is adopted, only a miniature probe can be adopted to perform two-dimensional scanning imaging on a living sample (an animal to be studied), the desktop multi-photon microscope comprises a femtosecond laser modulator and a miniature probe, and the miniature probe comprises: the scanning imaging part is used for receiving laser output by the femtosecond laser modulator, the laser scans tissues in the living body sample to excite fluorescent dye in the living body sample and generate a fluorescent signal, the micro-electromechanical scanner, the objective lens, the collimator, the dichroic mirror, the micro-electromechanical scanner and the collecting lens, the lens cone lens and the scanning lens are connected to the objective lens, and the whole structure is heavy in weight and large in size.

However, the current technical method has the following technical defects:

first, the micro probe in the prior art adopts two-dimensional scanning imaging, the imaging speed is slow (only 40 Hz), and in addition, the imaged image is inconvenient for observing the space condition of living sample data acquisition.

Second, in the use of such a desktop multiphoton microscope, the head of the living body specimen must be fixed on the test table at all times, and the living body specimen is under physical restraint and emotional stress (which is prone to fear) during the experiment, so that the behavior of the living body specimen in the case of free movement cannot be effectively studied.

Third, in the prior art, the fluorescent dye must be injected into the living body sample first, and this injection method cannot be used for the patient, which results in the failure of clinical application.

Fourth, such a desktop multiphoton microscope has a tube lens and a scanning lens, so that the size of the microscope cannot be further reduced.

SUMMERY OF THE UTILITY MODEL

The utility model provides a microscope to solve among the prior art desk-top many photon microscope and can only realize two-dimensional scanning formation of image, and this structure weight is heavy, bulky, still has the technical problem that can't be applied to clinically in addition.

In order to achieve the above object, the basic scheme of the present invention is as follows:

the three-dimensional line scanning optical imaging structure comprises a light source emitter and a collecting lens, wherein the light source emitter is a laser emitter, and a collimating lens, a cylindrical lens, a first scanner, a focusing lens, a wave plate, a second scanner and the collecting lens are sequentially arranged on a light path between the laser emitter and the collecting lens; the first scanner and the second scanner are both dichroic mirror scanners, the first scanner can rotate along the vertical direction of the light path, and the second scanner can move along the direction of the light path; the rear part of the first scanner is provided with an objective lens, and the first scanner reflects the laser from the cylindrical lens to a focusing lens in front of the first scanner and transmits the nonlinear light to the objective lens in the rear part of the first scanner; the cylindrical lens is used for focusing the laser collimated by the collimating lens into a linear focus on the surface of the first scanner.

The technical principle is as follows: when the scheme is adopted, laser emitted by a laser emitter is changed into parallel laser after passing through a collimating lens, the parallel laser is changed into linear laser after passing through a cylindrical lens, the linear laser is reflected into a focusing lens from a scanning unit, the linear laser is focused into a linear laser in the other direction, then the linear laser is polarized by a certain angle through a wave plate, the polarized linear laser reaches a scanner unit II and is reflected onto the wave plate by the scanner, the linear laser is polarized again by the wave plate, the linear laser rotates by a certain angle again, the linear laser reaches the scanner unit I after passing through the focusing lens again and is transmitted onto an objective lens from the scanner unit I, the objective lens converges the laser from the scanner unit I into a living body sample to excite the living body sample to generate a nonlinear optical signal, and the nonlinear optical signal is output to the scanner unit I, and the nonlinear optical signal sequentially passes through the focusing lens, And the wave plate and the scanner II finally reach the collecting lens to finish the collection of the image.

Rotating the scanner to scan the linear laser on one plane to form two-dimensional linear scanning of the living body sample; and after the two-dimensional line scanning is finished, moving the second scanner, enabling the two-dimensional line scanning plane of the internal tissue of the living body sample to move along the optical path direction through the far-end scanning principle, and further realizing the three-dimensional line scanning through the movement of the second scanner in the optical path direction.

Compare the beneficial effect in prior art:

first, in the aspect of the formation of image to living body sample data acquisition, scanning lens and tube lens are not set up to this scheme, replace dichroscope and micro-electromechanical system scanner among the prior art through dichroscope scanner to reach under the prerequisite that satisfies the imaging quality, improve imaging speed greatly, optimize inner structure, reduce the purpose of self weight.

Secondly, under the prerequisite of the weight reduction of optical imaging structure, when carrying out the data acquisition of live body sample, especially in the aspect of the animal brain formation of image that carries out the activity, more conveniently wear on the head of animal, reduce the influence of weight to animal activity, avoid detection error.

Thirdly, this scheme can realize three-dimensional line scanning, compares in adopting two-dimensional scanning to compare among the prior art, and three-dimensional scanning makes the data collection third dimension of live body sample stronger, is favorable to the research to the live body sample.

Fourthly, the line character laser is directly focused on the detection plane of the living body sample to excite the living body sample to generate a nonlinear optical signal, and the whole process does not need to use fluorescent dye for the living body sample, so that the subsequent application in clinic is facilitated.

Further, the dichroic mirror scanner comprises a dichroic mirror and a driver which does not affect transmission of nonlinear optical signals, the dichroic mirror is connected with the driver, the driver of the first scanner can drive the connected dichroic mirror to rotate along the vertical direction of the optical path, and the driver of the second scanner can drive the connected dichroic mirror to move along the direction of the optical path.

Has the advantages that: the movement or rotation of the dichroic mirror is achieved by means of a drive.

Further, the lenticular lens is a plano-convex lens.

Has the advantages that: the plano-convex lens can focus the laser collimated by the collimating lens into a linear focus on the surface of the first scanner.

Further, the rear part of the collecting lens is externally connected with a photoelectric imaging device.

The photoelectric imaging device can timely store the image data collected by the collecting lens, and in addition, the photoelectric imaging device is provided with a camera with a synchronous rolling exposure shutter technology, and the position of the linear laser is strictly synchronous with a certain row of photoelectric detection units read out by a rolling shutter of the camera, so that high-speed imaging is realized.

Further, the scanner further comprises a reflecting mirror, and the reflecting mirror is arranged on a light path between the collimating lens and the first scanner.

Has the advantages that: the design of adding the speculum can conveniently adjust the incident angle of the laser of input to scanner one, the formation of image of being convenient for.

Further, the objective lens adopts an aspheric objective lens.

Has the advantages that: the curvature radius of the aspheric objective lens is changed along with the central axis, so that the optical quality is improved, optical elements are reduced, and the design cost is reduced.

Further, the objective lens is an achromatic objective lens.

Further, the collimating lens is an achromatic collimating lens.

Further, the focusing lens is an achromatic focusing lens.

Has the advantages that: the objective lens or the collimating lens or the focusing lens with the achromatization function is adopted, which is beneficial to balancing aberration in a certain wave band range, and further improves the transmission efficiency, the light beam focusing condition and the excitation efficiency of the laser.

Further, a microscope comprising a body on which the three-dimensional line scanning optical imaging structure of any one of claims 1-9 is mounted.

Has the advantages that: the microscope using the three-dimensional line scanning optical imaging structure is further convenient for observing the data collected by the living body sample.

Drawings

FIG. 1 is a schematic top view of an embodiment of the present invention;

fig. 2 is an isometric view of a schematic structural diagram of an embodiment of the present invention;

fig. 3 is a schematic structural diagram of a fourth embodiment of the present invention.

Detailed Description

The following is further detailed by way of specific embodiments:

reference numerals in the drawings of the specification include: the device comprises a collimating lens 10, a cylindrical lens 12, a reflecting mirror 20, a first scanner 30, an objective lens 40, a focusing lens 50, a wave plate 60, a second scanner 70, a collecting lens 80, a laser emitter 90, a photoelectric imaging device 91, a main body 100 and a detection plane 110.

Wherein the X, Y and Z directions are perpendicular to each other two by two.

Example one

An embodiment substantially as shown in figures 1 and 2 of the accompanying drawings:

three-dimensional line scanning optical imaging structure includes according to the light path in proper order: the laser scanning device comprises a laser emitter 90, a collimating lens 10, a reflecting mirror 20, a cylindrical lens 12, a first scanner 30 rotating along an X axis, a focusing lens 50, a wave plate 60, a second scanner 70 moving along a Z axis direction and a collecting lens 80, wherein an objective lens 40 is arranged on the right side of the first scanner 30, and the first scanner 30 reflects laser light from the cylindrical lens 12 to the focusing lens 50 on the left side and transmits nonlinear light to the objective lens 40 on the right side.

And the laser emitter 90 is used for emitting laser with any wavelength between 700nm and 1600 nm.

The collimating lens 10 is used for collimating the laser light emitted from the laser emitter 90 (the laser light emitted from the laser emitter 90 is changed into a plurality of parallel laser lights after passing through the collimating lens 10), and the collimating lens 10 is a collimating lens 10 with achromatic function (for example, an achromatic collimating lens 10(#65-286, Edmund Optics inc., Barrington, NJ, USA; diameter: 2mm, equivalent focal length: 3mm, special near infrared light is used).

The reflector 20 is used for translating a light path, is made of optical glass or high polymer, and has an optical coating film for enhancing the transmissivity on a transmission surface and an optical coating film for enhancing the reflectivity on a reflection surface; the reflector 20 is used for adjusting the angle of the parallel laser output by the collimating lens 10 and reflecting the parallel laser to the cylindrical lens 12, the reflector 20 comprises a transmission surface and a reflection surface, the transmission surface is provided with an optical coating film for enhancing the transmissivity, and the reflection surface is provided with an optical coating film for enhancing the reflectivity; the mirror 20 is positioned at 45 degrees.

The cylindrical lens 12 is used for focusing the collimated parallel laser light into a linear laser focus in the X direction of the surface of the first scanner 30.

The first scanner 30 is used to project the nonlinear optical signal and separate the linear laser beam with polarization X, the first scanner 30 is schematically shown in figure 2,

the first scanner 30 and the second scanner 70 are both dichroic mirror scanners, each dichroic mirror scanner comprises a dichroic lens and an annular micro-electromechanical driver which does not affect transmission of nonlinear optical signals, the dichroic mirror covers the annular micro-electromechanical driver, the dichroic lens is made of optical glass or high polymer, the cut-off wavelength of the dichroic lens is 700nm, and the dichroic mirror is used for reflecting s-type polarized laser with the wavelength of 700nm-1600nm and transmitting p-type polarized laser with the wavelength of 700nm-1600nm and nonlinear optical signals with the wavelength of 350nm-700 nm; the mems of scanner one 30 can drive the attached dichroic mirror to rotate along the X-axis, and the mems of scanner two 70 can drive the attached dichroic mirror to move along the Z-axis.

An objective lens 40 made of optical glass or high molecular polymer and having an optical coating film with enhanced transmittance on the surface; the objective lens 40 adopts a refractive lens, a gradient index lens or a gradient index lens with a curved surface profile, and is used for focusing the line laser projected from the scanner-30 in the sample for external imaging collection, and since the sample is not a transparent object, the line laser is focused on the sample, and then a nonlinear optical signal is reflected from the sample, and finally the nonlinear optical signal is collected by the collecting lens 80.

The focusing lens 50 is made of optical glass or high molecular polymer, and has an optical coating with enhanced transmittance on the surface thereof, and is used for focusing and coupling the received nonlinear optical signal into the collecting lens 80.

The collection lens 80 feeds back the nonlinear optical signal to an externally connected photo-electric imaging device 91.

The specific implementation process is as follows:

two-dimensional line scanning:

the s-type linear polarized laser is output from the laser emitter 90 to the collimating lens 10, the collimating lens 10 collimates the received s-type linear polarized laser into parallel light (collimation processing), and reduces chromatic aberration between the lasers with different frequencies through the collimating lens 10 with achromatic function (achromatic processing), then the mirror 20 reflects the collimated s-type linear polarized laser and focuses the laser on the X direction of the dichroic mirror surface of the first scanner 30 into a linear shape through the cylindrical lens 12, the dichroic mirror of the first scanner 30 reflects the s-type linear polarized laser, then the focusing lens 50 collimates the s-type linear polarized laser on the X direction and focuses the laser on the Y direction into a linear shape, the s-type linear polarized laser continuously passes through the wave plate 60, rotates 45 degrees in the polarization direction of the s-type linear polarization after passing through the wave plate 60, and then the laser is focused on the dichroic mirror surface of the second scanner 70 in the Y direction, the second scanner 70 reflects laser, the reflected and diffused laser passes through the wave plate 60 again, the polarization direction of the laser rotates 45 degrees in the same direction again, and then the laser becomes p-type linearly polarized light (perpendicular to s-type linearly polarized light), the laser becomes an X-direction focused light beam through the focusing lens 50 again, the Y-direction collimated light beam is projected on the surface of the dichroic lens of the first scanner 30, the dichroic lens of the first scanner 30 transmits the p-type linearly polarized laser, the first scanner 30 is located on the back focal plane of the objective lens 40, and finally the p-type linearly polarized light forms a linear focus which is located in the sample and is collimated in the X direction and focused in the Y direction through the objective lens 40; when the dichroic mirror in the first scanner 30 is driven by the mems driver to rotate along the X-axis, the linear focus will also scan along the X-direction (the X-direction of the detection plane 110 shown in fig. 2), so as to form a two-dimensional scanning track, and thus, the laser can perform two-dimensional line scanning on the plane of the internal tissue of the living body sample.

Three-dimensional line scanning:

when the first scanner 30 finishes one frame of two-dimensional line Scanning image, the dichroic mirror on the second scanner 70 is moved along the Z direction by the micro-electro-mechanical driver, the two-dimensional line Scanning plane of the tissue inside the living body sample is also moved along the Z axis by the Remote Scanning principle (Remote Scanning, see Botcherby EJ, Smith CW, Kohl MM, et al. Abstract-free three-dimensional multiple imaging of the nuclear activity kHz rates. Proceedings of the national academy of Sciences of the United States of America. 2012; 109: 2919-2924.doi:10.1073/pnas. 1119.) to realize three-dimensional line Scanning by the movement of the second scanner 70 in the Z direction, the nonlinear signal is excited in the sample by the objective lens 40, the linear signal is collected by the linear lens 40, the linear signal is focused by the first linear lens 60, the dichroic mirror is focused by the second focusing lens 60, the dichroic mirror of scanner two 70 transmits the nonlinear signal wavelength and the nonlinear signal is then fed back by the collection lens 80 into an externally connected photo-electric imaging device 91.

In addition, due to the linear focus formed by the linear scanning method adopted in the present embodiment, the fluorescence collected by the objective lens 40 is also linear and moves in parallel in the external connected photoelectric imaging device 91 along with the rotation of the dichroic mirror in the scanner-30, the detection of the moving linear laser is performed by a scientific complementary metal oxide semiconductor (sCMOS) camera having a synchronizable rolling exposure shutter technology in the photoelectric imaging device 91, and the position of the linear laser is strictly synchronized with a certain line of photoelectric detection units currently read out by the rolling shutter of the sCMOS camera, thereby realizing high-speed imaging.

The imaging quality is met through the dichroic mirror scanner, the imaging speed is greatly improved, the internal structure is simple and small, and the weight of the whole three-dimensional line scanning optical imaging structure is small; when the data acquisition of living body sample, especially in the aspect of the animal brain formation of image that carries out the activity, more conveniently wear on the head of animal, reduce the influence of weight to animal activity, avoid detection error.

The three-dimensional scanning is adopted in the embodiment, so that the data collection stereoscopic impression of the living body sample is stronger, the research on the living body sample is facilitated, the fluorescent dye is not required to be used for the living body sample in the whole process, and the follow-up clinical application is facilitated.

Example two

The difference between the second embodiment and the first embodiment is that the cylindrical lens 12 is a plano-convex lens, the objective lens 40 is an achromatic objective lens 40, the design wavelength of the objective lens 40 is any 2 wavelengths between 700nm and 1600nm, the second embodiment is 817nm and 1064nm (but not limited to these two wavelengths), the material is optical glass or polymer, the surface has an optical coating for enhancing transmittance, and the structure can be a conventional refractive lens, a gradient index lens or a gradient index lens with a curved surface profile, and is used for focusing incident laser in an external sample, exciting a nonlinear optical signal, and collecting the nonlinear optical signal in an epi-detection manner.

EXAMPLE III

The difference between the third embodiment and the first embodiment is that the focusing lens 50 is an achromatic focusing lens 50, the design wavelength is any 2 wavelengths between 350nm and 700nm, the third embodiment uses wavelengths of 408nm and 633nm (but not limited to these two wavelengths), the material is optical glass or high molecular polymer, and the surface has an optical coating film with enhanced transmittance.

Example four

Referring to fig. 3, a microscope includes a main body 100, and the three-dimensional line scanning optical imaging structure according to the first embodiment is mounted on the main body 100.

The microscope with the three-dimensional line scanning optical imaging structure is used, so that the data collected by the living body sample can be observed conveniently.

The above description is only an example of the present invention, and the common general knowledge of the known specific structures and characteristics of the embodiments is not described herein. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several modifications and improvements can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.

Claims

1. A microscope, comprising a body, characterized in that: the three-dimensional line scanning optical imaging structure comprises a light source emitter and a collecting lens, the light source emitter is a laser emitter, and a collimating lens, a cylindrical lens, a first scanner, a focusing lens, a wave plate, a second scanner and the collecting lens are sequentially arranged on a light path between the laser emitter and the collecting lens;

the first scanner and the second scanner are both dichroic mirror scanners, the first scanner can rotate along the vertical direction of the light path, and the second scanner can move along the direction of the light path;

the rear part of the first scanner is provided with an objective lens, and the first scanner reflects the laser from the cylindrical lens to a focusing lens in front of the first scanner and transmits the nonlinear light to the objective lens in the rear part of the first scanner;

the cylindrical lens is used for focusing the laser collimated by the collimating lens into a linear focus on the surface of the first scanner;

the rear part of the collecting lens is externally connected with a photoelectric imaging device.

2. A microscope according to claim 1, wherein: the dichroic mirror scanner comprises a dichroic mirror and a driver which does not affect transmission of nonlinear optical signals, the dichroic mirror is connected with the driver, the driver of the first scanner can drive the connected dichroic mirror to rotate along the vertical direction of a light path, and the driver of the second scanner can drive the connected dichroic mirror to move along the direction of the light path.

3. A microscope according to claim 2, wherein: the cylindrical lens is a plano-convex lens.

4. A microscope according to claim 1, wherein: the scanner further comprises a reflecting mirror, and the reflecting mirror is arranged on a light path between the collimating lens and the first scanner.

5. A microscope according to claim 1, wherein: the objective lens adopts an aspheric objective lens.

6. A microscope according to claim 1, wherein: the objective lens is an achromatic objective lens.

7. A microscope according to claim 1, wherein: the collimating lens is an achromatic collimating lens.

8. A microscope according to claim 1, wherein: the focusing lens is an achromatic focusing lens.