CN112858238A - Micro-droplet fluorescence signal detection device comprising optical fiber - Google Patents
Micro-droplet fluorescence signal detection device comprising optical fiber Download PDFInfo
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- CN112858238A CN112858238A CN202110059076.1A CN202110059076A CN112858238A CN 112858238 A CN112858238 A CN 112858238A CN 202110059076 A CN202110059076 A CN 202110059076A CN 112858238 A CN112858238 A CN 112858238A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6484—Optical fibres
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Abstract
The invention provides a micro-droplet fluorescence signal detection device comprising an optical fiber, which comprises a light combination module, a light source module and a light source module, wherein the light combination module is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed excitation light; an objective lens on a light conduction path of the mixed excitation light; the light processing module comprises a convex lens for focusing the first fluorescence and the second fluorescence to pass through, a first dichroic mirror for reflecting the mixed excitation light to the objective lens from the light combining module, and an optical fiber, wherein the first end of the optical fiber is positioned on a focus of the convex lens at one side far away from the objective lens so as to receive and transmit the first fluorescence and the second fluorescence focused by the convex lens; and the signal receiving module is connected to the second end of the optical fiber and used for processing the fluorescent signal received and transmitted by the optical fiber. The optical fiber is used as a rear-end fluorescence collecting and transmitting component, so that the loss of fluorescence signals is effectively reduced, the optical processing module and the signal receiving module are respectively and independently connected through the optical fiber, the signal receiving module can be randomly arranged in space, and the flexibility of system arrangement is improved.
Description
Technical Field
The invention belongs to the technical field of microfluidic detection chips, and particularly relates to a micro-droplet fluorescence signal detection device comprising an optical fiber.
Background
The biochip has wide application in new medicine development, disease diagnosis, gene expression analysis, etc. The technology of the microfluidic detection chip is mature day by day and becomes a focus of people. There are various biological and chemical processes in the microfluidic detection chip, and the processes are usually completed in a micro-scale flow channel space, in which some devices capable of detecting the reaction process are required.
Traditional confocal detection device of laser, the interference of stray light outside the detection light path for eliminating the focal plane, improve detectivity, set up the pinhole at the receiving terminal, its spatial structure mode is single and fixed, no matter be single laser, double laser or many laser arouse signal detection device and all have the fixed increase of pinhole quantity, the laser quantity that is used for arousing the light increases the optical path increase, detection device follows the shortcoming such as laser to collect that signal end wholeness requires height, the degree of difficulty increases when leading to the debugging because the increase of pinhole quantity, the signal reception error of device has been increased in the intangible, carry out multichannel signal detection time measuring, because space optical path increases and leads to partly signal attenuation and loss, corresponding detected signal existence is lost, the wholeness requires high equipment and the later maintenance that is not convenient for the product.
Disclosure of Invention
Therefore, the technical problem to be solved by the present invention is to provide a micro-droplet fluorescence signal detection apparatus including an optical fiber, wherein the optical fiber is used as a back-end fluorescence collection and transmission component, so as to effectively reduce the loss of fluorescence signals, and meanwhile, the optical processing module and the signal receiving module are respectively and independently connected through the optical fiber, so that the signal receiving module can be arbitrarily arranged in space, thereby improving the flexibility of system arrangement.
In order to solve the above problems, the present invention provides a micro-droplet fluorescence signal detection device including an optical fiber, including: the light combining module is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed exciting light;
the objective lens is positioned on a light conduction path of the mixed exciting light, and can focus the mixed exciting light on the micro-droplet to be detected so as to excite and generate first fluorescence corresponding to the first laser and second fluorescence corresponding to the second laser;
the light processing module comprises a convex lens for focusing the first fluorescence and the second fluorescence to pass through, a first dichroic mirror for reflecting the mixed excitation light to the objective lens from the light combining module, and an optical fiber, wherein the first end of the optical fiber is positioned on a focus of the convex lens at one side far away from the objective lens so as to receive and transmit the first fluorescence and the second fluorescence focused by the convex lens;
and the signal receiving module is connected to the second end of the optical fiber and used for processing the first fluorescence and the second fluorescence received and transmitted by the optical fiber.
Preferably, the light processing module further comprises a light processing housing, the light processing housing comprises a fixing piece, a first connecting piece and a second connecting piece, the fixing piece and the second connecting piece clamp and position the convex lens, the first connecting piece and the second connecting piece clamp and position the first dichroic mirror, the fixing piece is further connected with a first optical fiber adapter, and the first optical fiber adapter can position the first end of the optical fiber.
Preferably, the light processing housing further comprises a sealing cover, and the sealing cover is connected with the fixing piece and covers the outer side of the first optical fiber adapter; and/or a first dichroic mirror rubber pad is further clamped between the first connecting piece and the second connecting piece.
Preferably, the second connecting piece with still include the convex lens lantern ring between the mounting, convex lens is in the central through-hole of the convex lens lantern ring, the convex lens lantern ring is right with the clamping ring convex lens forms the centre gripping.
Preferably, the signal receiving module includes a signal receiving housing, the signal receiving housing includes a first main body, a second main body, a first connection seat, a second connection seat, the first main body is connected to the second end of the optical fiber through a second optical fiber adapter, a second dichroic mirror is sandwiched between the first main body and the second main body, a first color filter is sandwiched between the second main body and the first connection seat, a second color filter is sandwiched between the first main body and the second connection seat, a first photomultiplier is connected to the first connection seat to process the first fluorescence received and transmitted through the optical fiber, a second photomultiplier is connected to the second connection seat to process the second fluorescence received and transmitted through the optical fiber, and a collimating assembly is sandwiched between the second optical fiber adapter and the first main body, so as to refract the scattered light of the light received and transmitted by the optical fiber into parallel light.
Preferably, the collimating assembly includes a collimating lens barrel and a collimating lens located therein, and the collimating lens is held by the collimating lens barrel and the collimating lens holder.
Preferably, a diaphragm is further disposed on a light incident side of the collimating lens, and the diaphragm is located in the collimating lens barrel.
Preferably, the light combining module includes a first laser, a second laser, a reflective mirror and a third dichroic mirror, and the first laser light emitted by the first laser is reflected by the reflective mirror and then combined with the second laser light emitted by the second laser into the mixed excitation light at the third dichroic mirror.
Preferably, the light combining module further includes a fixing plate and a light combining cassette, the first laser, the second laser and the light combining cassette are fixedly connected to the fixing plate, the reflective mirror and the third dichroic mirror are disposed in the light combining cassette, and the light combining cassette has a first laser entrance port, a second laser entrance port and a mixed excitation light exit port.
Preferably, the position of the objective lens can be adjusted by driving a focusing motor.
According to the micro-droplet fluorescence signal detection device comprising the optical fiber, the optical fiber is used as a rear-end fluorescence collection and transmission component, so that the loss of fluorescence signals (namely the first fluorescence and the second fluorescence) is effectively reduced, and meanwhile, the optical processing module and the signal receiving module are respectively and independently arranged and are connected through the optical fiber, so that the signal receiving module can be randomly arranged in space, and the flexibility of system arrangement is improved.
Drawings
FIG. 1 is a schematic diagram of a micro-droplet fluorescence signal detection device including an optical fiber according to an embodiment of the present invention (arrows indicate light transmission paths);
FIG. 2 is a schematic structural diagram of a micro-droplet fluorescence signal detection device including an optical fiber according to an embodiment of the present invention;
FIG. 3 is a schematic view of the disassembled structure of the optical processing module in FIG. 2;
fig. 4 is a disassembled structural diagram of the signal receiving module in fig. 2;
fig. 5 is a schematic view of a disassembled structure of the light combining module in fig. 2.
The reference numerals are represented as:
1. a light combining module; 11. a first laser; 12. a second laser; 13. a fixing plate; 14. a light-combining cassette; 141. a reflective mirror; 142. a third dichroic mirror; 2. an objective lens; 21. a focus motor; 3. a light processing module; 31. a first dichroic mirror; 311. a first dichroic mirror rubber pad; 32. a convex lens; 321. a convex lens collar; 322. a lens rubber pad; 323. pressing a ring; 33. an optical fiber; 341. a fixing member; 342. a first connecting member; 343. a second connecting member; 344. a first optical fiber adapter; 345. a sealing cover; 346. a light extinction device; 4. a signal receiving module; 41. a first photomultiplier tube; 42. a second photomultiplier tube; 43. a diaphragm; 431. a diaphragm fixing member; 44. a collimating lens; 441. a collimator lens fixing seat; 45. a second dichroic mirror; 46. a second color filter; 47. a first color filter; 481. a second optical fiber adapter; 482. a collimating lens barrel; 483. a first body member; 484. a second body member; 485. a first fastening ring; 486. a second fastening ring; 487. a first connecting seat; 488. a second connecting seat; 491. a second dichroic mirror rubber pad; 492. a color filter rubber pad; 493. photomultiplier rubber pad; 100. micro-droplets are detected.
Detailed Description
Referring to fig. 1 to 5 in combination, according to an embodiment of the present invention, there is provided a microdroplet fluorescence signal detection device including an optical fiber, including: the light combining module 1 is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed excitation light; the objective lens 2 is located on a light conduction path of the mixed excitation light, and can focus the mixed excitation light on the microdroplet 100 to be detected so as to excite and generate a first fluorescence corresponding to the first laser and a second fluorescence corresponding to the second laser, and in order to achieve the purpose of focusing, the position of the objective lens 2 can be adjusted under the driving of a focusing motor 21, specifically, the focusing motor 21 can be a fixed shaft motor, and when the focusing motor 21 operates, the fixed shaft motor generates axial displacement, so that the position of the objective lens 2 is adjusted; the light processing module 3 comprises a convex lens 32 through which the first fluorescence and the second fluorescence are focused, a first dichroic mirror 31 for reflecting the mixed excitation light from the light combining module 1 to the objective lens 2, and an optical fiber 33, wherein a first end of the optical fiber 33 is located at a focal point on a side of the convex lens 32 away from the objective lens 2, so as to receive and transmit the first fluorescence and the second fluorescence focused by the convex lens 32; and the signal receiving module 4 is connected to the second end of the optical fiber 33 and is used for processing the first fluorescence and the second fluorescence received and transmitted by the optical fiber 33. In the technical scheme, the optical fiber 33 is used as a rear-end fluorescence collecting and transmitting component, so that the loss of fluorescence signals (namely the first fluorescence and the second fluorescence) is effectively reduced, and meanwhile, the optical processing module 3 and the signal receiving module 4 are respectively and independently arranged and connected through the optical fiber 33, so that the signal receiving module 4 can be randomly arranged in space, and the flexibility of system arrangement is improved.
As a specific implementation manner, preferably, the optical processing module 3 further includes a light processing housing, the light processing housing includes a fixing member 341, a first connecting member 342, and a second connecting member 343, the fixing member 341 and the second connecting member 343 clamp and position the convex lens 32, the first connecting member 342 and the second connecting member 343 clamp and position the first dichroic mirror 31, the fixing member 341 is further connected with a first optical fiber adapter 344, and the first optical fiber adapter 344 can position the first end of the optical fiber 33. In this technical solution, the fixing member 341, the first connecting member 342, and the second connecting member 343, which are connected to each other as a whole, form the light processing housing (two adjacent components may be bolted, for example), and at the same time, the first dichroic mirror 31 and the convex lens 32 are reliably clamped by the two adjacent components, so that the light processing module 3 is more reasonable and compact in structure. The first fiber adapter 344 is configured to position the first end of the optical fiber 33 more accurately and reliably.
Further, the optical processing housing further includes a sealing cover 345, where the sealing cover 345 is connected to the fixing member 341 and covers the outer side of the first optical fiber adapter 344 to seal the first end of the optical fiber 33, so as to prevent external stray light from interfering with the optical signal received and transmitted in the optical fiber 33; and/or a first dichroic mirror rubber pad 311 is further sandwiched between the first connecting piece 342 and the second connecting piece 343, so as to form protection and light-tight for the first dichroic mirror 31.
In some embodiments, a convex lens collar 321 is further included between the second connecting member 343 and the fixing member 341, and is configured to form a positioning support for the convex lens 32, specifically, the convex lens 32 is located in a central through hole of the convex lens collar 321, the convex lens collar 321 and the pressing ring 323 form a clamping for the convex lens 32, and further, a lens rubber pad 322 is further provided between the pressing ring 323 and the convex lens 32 to form a protection for the convex lens 32.
As a specific embodiment, it is preferable that the signal receiving module 4 includes a signal receiving housing, the signal receiving housing includes a first main body piece 483, a second main body piece 484, a first connecting seat 487, a second connecting seat 488, the first main body piece 483 is connected to the second end of the optical fiber 33 through a second optical fiber adapter 481, a second dichroic mirror 45 and a second dichroic rubber pad 491 are sequentially interposed between the first main body piece 483 and the second main body piece 484, a first color filter 47, a color filter rubber pad 492, a first fastening ring 485 and a photomultiplier rubber pad 493 are sequentially interposed between the second main body piece 484 and the first connecting seat 487, a second color filter 46, a rubber pad 492, a second fastening ring 486 and a photomultiplier rubber pad 493 are sequentially interposed between the first main body piece 483 and the second connecting seat 488, the first connection seat 487 is connected to a first photomultiplier 41 to process the first fluorescent light received and transmitted through the optical fiber 33, the second connection seat 488 is connected to a second photomultiplier 42 to process the second fluorescent light received and transmitted through the optical fiber 33, and a collimating assembly is further interposed between the second optical fiber adapter 481 and the first main body 483 to refract the scattered light of the light received and transmitted through the optical fiber 33 into parallel light. In the technical scheme, the photomultiplier (i.e. the first photomultiplier 41 and the second photomultiplier 42) is used for conducting the acquired data to corresponding data processing equipment (e.g. a computer) for corresponding processing (e.g. counting), so that a plurality of detection parameters can be acquired by single detection, the detection result is richer, the applicable scenes and fields are wider, and the experiment cost can be greatly reduced and the detection work efficiency is improved.
Further, the collimating assembly includes a collimating lens barrel 482 and a collimating lens 44 disposed therein, and a collimating lens holder 441, the collimating lens barrel 482 and the collimating lens holder 441 form a clamp for the collimating lens 44, preferably, a diaphragm 43 (which may be fixed in the collimating lens barrel 482 by a diaphragm fixing member 431) is further disposed on a light incident side of the collimating lens 44, and the diaphragm 43 is disposed in the collimating lens barrel 482, and the diaphragm 43 can eliminate stray light transmitted from the light processing module 3, and only a fluorescent signal is retained to pass through the collimating lens 44, so that the stray light passing through the collimating lens 44 is refracted into parallel light.
In some embodiments, the light combining module 1 includes a first laser 11, a second laser 12, a reflective mirror 141, and a third dichroic mirror 142, and the first laser light emitted from the first laser 11 is reflected by the reflective mirror 141 and combined with the second laser light emitted from the second laser 12 into the mixed excitation light by the third dichroic mirror 142, and in a specific structural aspect, the light combining module 1 further includes a fixing plate 13 and a light combining cassette 14, the first laser 11, the second laser 12, and the light combining cassette 14 are fixedly connected to the fixing plate 13, the reflective mirror 141 and the third dichroic mirror 142 are disposed in the light combining cassette 14, and the light combining cassette 14 has a first laser light incident port, a second laser light incident port, and a mixed excitation light emitting port. As a specific embodiment, the first laser may be a 532nm laser and the second laser may be a 473nm laser.
It is readily understood by a person skilled in the art that the advantageous ways described above can be freely combined, superimposed without conflict.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention. The above is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several improvements and modifications can be made without departing from the technical principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.
Claims (10)
1. A microdroplet fluorescence signal detection device comprising an optical fiber, comprising:
the light combining module (1) is used for combining first laser with a first wavelength and second laser with a second wavelength into mixed excitation light;
the objective lens (2) is positioned on a light conduction path of the mixed exciting light, and can focus the mixed exciting light on the micro-droplet (100) to be detected so as to excite and generate first fluorescence corresponding to the first laser and second fluorescence corresponding to the second laser;
the light processing module (3) comprises a convex lens (32) through which the first fluorescence and the second fluorescence are focused, a first dichroic mirror (31) for reflecting the mixed excitation light from the light combining module (1) to the objective lens (2), and an optical fiber (33), wherein a first end of the optical fiber (33) is positioned at a focus of the convex lens (32) on the side away from the objective lens (2) so as to receive and transmit the first fluorescence and the second fluorescence focused by the convex lens (32);
and the signal receiving module (4) is connected to the second end of the optical fiber (33) and is used for processing the first fluorescence and the second fluorescence received and transmitted by the optical fiber (33).
2. The optical fiber-containing micro droplet fluorescence signal detection device according to claim 1, wherein the optical processing module (3) further comprises an optical processing housing, the optical processing housing comprises a fixing member (341), a first connecting member (342) and a second connecting member (343), the fixing member (341) and the second connecting member (343) form a clamping position for the convex lens (32), the first connecting member (342) and the second connecting member (343) form a clamping position for the first dichroic mirror (31), the fixing member (341) is further connected with a first optical fiber adapter (344), and the first optical fiber adapter (344) can form a positioning for the first end of the optical fiber (33).
3. The detection apparatus for fluorescence signal of microdroplet containing optical fiber according to claim 2, wherein the light processing housing further comprises a sealing cover (345), the sealing cover (345) is connected with the fixing member (341) and covers the outside of the first optical fiber adapter (344); and/or a first dichroic mirror rubber pad (311) is further clamped between the first connecting piece (342) and the second connecting piece (343).
4. The optical fiber-containing micro droplet fluorescence signal detection device according to claim 2, further comprising a convex lens collar (321) between the second connector (343) and the fixing member (341), wherein the convex lens (32) is located in a central through hole of the convex lens collar (321), and the convex lens collar (321) and the pressing ring (323) clamp the convex lens (32).
5. The apparatus for detecting the fluorescence signal of the microdroplet containing the optical fiber according to claim 1, wherein the signal receiving module (4) comprises a signal receiving housing, the signal receiving housing comprises a first main body piece (483), a second main body piece (484), a first connecting seat (487), a second connecting seat (488), the first main body piece (483) is connected with the second end of the optical fiber (33) through a second optical fiber adapter (481), a second dichroic mirror (45) is sandwiched between the first main body piece (483) and the second main body piece (484), a first color filter (47) is sandwiched between the second main body piece (484) and the first connecting seat (487), a second color filter (46) is sandwiched between the first main body piece (483) and the second connecting seat (488), and a first photomultiplier (41) is connected to the first connecting seat (487) to clamp the first photomultiplier (41) received and transmitted through the optical fiber (33) The fluorescence is processed, a second photomultiplier (42) is connected to the second connecting seat (488) to process the second fluorescence received and transmitted through the optical fiber (33), and a collimation assembly is further clamped between the second optical fiber adapter (481) and the first main body piece (483) to refract scattered light of the light received and transmitted by the optical fiber (33) into parallel light.
6. The apparatus for detecting fluorescence signal of microdroplet containing optical fiber according to claim 5, wherein the collimating assembly comprises a collimating lens barrel (482) and a collimating lens (44) therein, and a collimating lens holder (441), wherein the collimating lens barrel (482) and the collimating lens holder (441) clamp the collimating lens (44).
7. The optical fiber-containing micro-droplet fluorescence signal detection device according to claim 6, wherein the light incident side of the collimating lens (44) is further provided with a diaphragm (43), and the diaphragm (43) is located in the collimating lens barrel (482).
8. The apparatus for detecting a fluorescence signal of a microdroplet containing an optical fiber according to claim 1, wherein the light combining module (1) comprises a first laser (11), a second laser (12), a reflective mirror (141) and a third dichroic mirror (142), and the first laser light emitted from the first laser (11) is reflected by the reflective mirror (141) and then combined with the second laser light emitted from the second laser (12) into the mixed excitation light by the third dichroic mirror (142).
9. The apparatus for detecting the fluorescence signal of the microdroplet containing the optical fiber according to claim 8, wherein the light combining module (1) further comprises a fixing plate (13) and a light combining cassette (14), the first laser (11), the second laser (12) and the light combining cassette (14) are fixedly connected to the fixing plate (13), the reflective mirror (141) and the third dichroic mirror (142) are disposed in the light combining cassette (14), and the light combining cassette (14) has a first laser incident port, a second laser incident port and a mixed excitation light exit port.
10. The optical fiber-containing micro droplet fluorescence signal detecting device according to claim 1, wherein the position of the objective lens (2) can be adjusted by driving a focusing motor (21).
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| CN202110059076.1A CN112858238A (en) | 2021-01-17 | 2021-01-17 | Micro-droplet fluorescence signal detection device comprising optical fiber |
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2021
- 2021-01-17 CN CN202110059076.1A patent/CN112858238A/en active Pending
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| US20010006434A1 (en) * | 1999-11-30 | 2001-07-05 | Yoo Jang-Hoon | Objective lens for high-density optical focusing and an optical disk in an optical pickup |
| US20100084785A1 (en) * | 2008-10-02 | 2010-04-08 | Sony Corporation | Method for manufacturing master and method for manufacturing optical disc |
| CN108414442A (en) * | 2018-02-09 | 2018-08-17 | 浙江大学 | Confocal microscope system suitable for near-infrared 2nd area fluorescent vital imaging |
| CN108956561A (en) * | 2018-06-07 | 2018-12-07 | 浙江大学 | Copolymerization coke and annular total internal reflection double mode microscopic system based on scanning galvanometer |
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