CN100427926C - Biochip imaging method splitted with laser cofocus scanning combined image and its device - Google Patents

Biochip imaging method splitted with laser cofocus scanning combined image and its device Download PDF

Info

Publication number
CN100427926C
CN100427926C CNB2005100970028A CN200510097002A CN100427926C CN 100427926 C CN100427926 C CN 100427926C CN B2005100970028 A CNB2005100970028 A CN B2005100970028A CN 200510097002 A CN200510097002 A CN 200510097002A CN 100427926 C CN100427926 C CN 100427926C
Authority
CN
China
Prior art keywords
biochip
flat
field objective
laser
subarea
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CNB2005100970028A
Other languages
Chinese (zh)
Other versions
CN1793860A (en
Inventor
王立强
吴碧珍
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ZHEJIANG JINCHENG SCI-TECH DEVELOPMENT Co Ltd
Zhejiang University ZJU
Original Assignee
ZHEJIANG JINCHENG SCI-TECH DEVELOPMENT Co Ltd
Zhejiang University ZJU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by ZHEJIANG JINCHENG SCI-TECH DEVELOPMENT Co Ltd, Zhejiang University ZJU filed Critical ZHEJIANG JINCHENG SCI-TECH DEVELOPMENT Co Ltd
Priority to CNB2005100970028A priority Critical patent/CN100427926C/en
Publication of CN1793860A publication Critical patent/CN1793860A/en
Application granted granted Critical
Publication of CN100427926C publication Critical patent/CN100427926C/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Abstract

The present invention discloses a biochip imaging method combining laser cofocus scan with image split joint. The present invention also discloses a device thereof. The whole biochip area is divided into a plurality of subareas, one-dimensional scan of each subarea is realized by optical scan composed of a vibration lens and a planar field objective lens, and the other dimensional scan is realized by mechanical scan composed of a motor and a guide rail. Fluorescent signals are converted to electric signals by a photoelectric detector and are collected by a computer to rebuild fluorescent images of the present subareas. After the first subarea is scanned to be imaged, the biochip translates a distance along the optical scanning direction to cause the next subarea to enter the optical scanning range so as to realize that the subarea is scanned to be imaged. In this way, after the last subarea is scanned to be imaged, the fluorescent image of every subarea is in split joint by the computer in order to synthesize a complete biochip fluorescent image. The present invention overcomes the shortages of long focus and small numerical aperture of the planar field objective lens in an optical scanner and enhances the optical resolution and the fluorescent detecting sensitivity.

Description

The biochip imaging method of splitted with laser cofocus scanning combined image and device thereof
Technical field
The present invention relates to fluorescence biosensor chip formation method and device thereof, especially the biochip imaging method of splitted with laser cofocus scanning combined image and device thereof.
Background technology
Fluorescence biosensor chip formation method based on laser cofocus scanning can be divided into two big classes according to scan mode: the first kind is that catoptron and small field of view object lens are fixed in the scanner head, and scanner head scans biochip fast along a certain direction; Second class is to adopt big visual field object lens, and laser beam is incident to object lens with different angles, is converted to along the linear sweep of a certain direction of biochip by the angle scanning of object lens with laser beam.United States Patent (USP) 5,459,325 and 6,407,858 have all described first kind scanister, and the former is fixed on catoptron and microcobjective in the scanner head, and this scanner head is by linear motor driving, along a quick to-and-fro movement of line slideway, thereby realization is to the rectilinear scanning of a direction of biochip; The latter is designed to the camber line scanning motion with scanner head, and catoptron and object lens are fixed on the oscillating arms of doing to swing with certain frequency, rotate around its axis, and therefore the original fluoroscopic image that obtains is the spherical coordinates image.The advantage of first kind scanister is that the focal length of object lens is short, numerical aperture is big, so optical resolution and phosphor collection efficient height, scanning field of view is all the time along optical axis direction simultaneously, object lens are gone up the image quality of point as long as guarantee axle, aberration to off-axis point does not require, and design of Optical System is simple; Therefore its shortcoming is that the weight of scanner head is bigger, scans that inertia is big, vibrations are big, and sweep velocity only about 20Hz, has not only reduced system reliability, and the analysis time of biochip is longer.For overcoming above-mentioned shortcoming, United States Patent (USP) 5,381,224 and 5,646,411 have described the second class scanister, the former makes up two galvanometers according to certain swing mode, the laser beam that makes incident realizes the two-dimensional scan of laser beam to biochip by big visual field object lens after vibration mirror reflected; The latter adopts a galvanometer, and galvanometer is being rotatablely moved in the swing fast again, and the laser beam of incident is reflexed to big visual field object lens with different angles, realizes the two-dimensional scan of biochip.Chinese patent 02112040.4 has been described galvanometer and has been realized jointly that with big visual field object lens the one-dimensional high-speed of biochip scans, and the step motor drive precise guide rail realizes another dimension low-velocity scanning of biochip, also belongs to the second class scanister.The major advantage of the second class device is that the moment of inertia of galvanometer is little, noise is little, sweep velocity can reach more than the 50Hz; Shortcoming mainly is that big visual field object lens must guarantee an image quality that goes up point and off-axis point simultaneously, the optical design complexity, and focal length is long, and numerical aperture is little, optical resolution that very difficult acquisition is very high and phosphor collection efficient.
Summary of the invention
The biochip imaging method and the device thereof that the purpose of this invention is to provide the splitted with laser cofocus scanning combined image that a kind of sweep velocity is fast, noise is little, optical resolution is high, phosphor collection efficient is high.
For reaching above-mentioned purpose, the biochip imaging method of splitted with laser cofocus scanning combined image of the present invention adopts scanning of flat-field objective subregion and image mosaic to realize the fluorescence imaging of whole biochip.
Step is as follows:
1) biochip is placed on the platform that moves by two driven by motor work two dimensions;
2) laser that laser instrument is sent expands bundle through beam expanding lens, successively through arriving galvanometer after mirror reflects and the dichroscope transmission around the axle swing, make laser beam reflex to flat-field objective with different angles, flat-field objective is with the laser beam transverse focusing of the different incidence angles degree lower surface at biochip, to the biochip transversal scanning, simultaneously, the lengthwise movement of first driven by motor platform, to the biochip longitudinal scanning, form the two-dimensional scan subregion of biochip;
3) fluorescence that sends of biochip is collected by flat-field objective, successively through galvanometer, dichroscope reflection, after the transmission of fluorescence color filter, arrive convergent mirror, on the focal plane of convergent mirror, place the confocal diaphragm that has aperture, make the fluorescence of biochip lower surface pass aperture and arrive photodetector, the output signal of photodetector is through advance signal processing unit processing and amplifying, by computer acquisition and rebuild the fluoroscopic image of this subregion;
4) second driven by motor platform transverse translation is to biochip next son zone repeating step 2) and 3), obtaining each sub regions image, each the sub regions image by obtaining in the computing machine splicing flat-field objective visual field obtains complete biological chip fluorescent picture.
The device that is used for above-mentioned biochip imaging method, comprise laser instrument, beam expanding lens, catoptron, dichroscope, galvanometer, flat-field objective, fluorescence color filter, convergent mirror, have confocal diaphragm, the photodetector of aperture and be used to place the platform of biochip, the screw mandrel that is provided with guide rail and guide rail, guide rail in the platform bottom links to each other with first motor shaft, the screw mandrel of guide rail links to each other with second motor shaft; Laser instrument, beam expanding lens, catoptron, dichroscope, galvanometer, flat-field objective are arranged in order, and form the laser scanning light path of biochip, and galvanometer is positioned at the front focal plane of flat-field objective, and biochip is positioned at the back focal plane of flat-field objective; On dichroiscopic reflected light path, place color filter, convergent mirror, confocal diaphragm and photodetector successively along the light reflection direction, confocal diaphragm is positioned at the back focal plane of convergent mirror.
The invention has the beneficial effects as follows: the focal length of small field of view flat-field objective is short, numerical aperture is big, volume is little, in the optical resolution and phosphor collection efficient that improve the biochip scanning imaging, has reduced the size of imaging device, has reduced system cost.
For instance, original 1/4 if the scanning field of view of flat-field objective is decreased to, then its focal length can reduce more than 1 times, and numerical aperture can increase 1 times, so optical resolution and phosphor collection efficient can improve more than 2 times.Whole imaging device can be selected littler galvanometer size, and the sweep velocity of galvanometer is faster, but reach 200Hz, though need this moment to divide to scan whole biochip 4 times, overall sweep velocity still can guarantee 50Hz.
The present invention has overcome the shortcoming of first kind device and the second class device in the background technology, and integrated advantage separately can be widely used in all kinds of fluorescent microscopic imaging system.
Description of drawings
Fig. 1 is that the present invention constitutes synoptic diagram.
Embodiment
With reference to Fig. 1, the device of the biochip imaging method of splitted with laser cofocus scanning combined image, comprise laser instrument 1, beam expanding lens 2, catoptron 3, dichroscope 4, galvanometer 5, flat-field objective 6, fluorescence color filter 13, convergent mirror 14, have confocal diaphragm 15, the photodetector 16 of aperture and be used to place the platform 8 of biochip 7, the screw mandrel that is provided with guide rail 9 and guide rail 10, guide rail 9 in platform 8 bottoms links to each other with 11 in first motor, the screw mandrel of guide rail 10 links to each other with 12 in second motor; Laser instrument 1, beam expanding lens 2, catoptron 3, dichroscope 4, galvanometer 5, flat-field objective 6 are arranged in order, and form the laser scanning light path of biochip 7, and galvanometer 5 is positioned at the front focal plane of flat-field objective 6, and biochip 7 is positioned at the back focal plane of flat-field objective 6; On the reflected light path of dichroscope 4, place color filter 13, convergent mirror 14, confocal diaphragm 15 and photodetector 16 successively along the light reflection direction, confocal diaphragm 15 is positioned at the back focal plane of convergent mirror 14.
During use, the photodetector 16 of device is continuous with the advance signal processing unit 17 that is connected computing machine 18, and advance signal processing unit 17 can be the OPA27 chip.Contrive equipment work is as follows: the laser that is sent by laser instrument 1 at first expands bundle through beam expanding lens 2, successively through arriving galvanometer 5 after catoptron 3 reflections and dichroscope 4 transmissions, galvanometer 5 is swung fast around its 5-1, laser beam is reflexed to flat-field objective 6 with different angles, flat-field objective 6 with the laser beam of different incidence angles degree along the diverse location of Y direction focusing at biochip 7 lower surfaces, excite the fluorescent marker of relevant position, realize the transversal scanning of biochip 7.First motor 11 promotes guide rail 9 and moves along directions X, realizes the longitudinal scanning of biochip 7.The visual field size of flat-field objective 6 only is the part of biochip width, therefore only scans biochip 7 one sub regions 7-1.The fluorescence that subregion 7-1 analyzing spot sends is collected by flat-field objective 6, successively through after galvanometer 5 reflections, dichroscope 4 reflections, 13 transmissions of fluorescence color filter, arrive convergent mirror 14, on the focal plane of convergent mirror 14, place the confocal diaphragm 15 that has aperture, only the fluorescence of biochip lower surface can pass aperture and arrive photodetector 16, the output signal of photodetector 16 is gathered and is rebuild the fluoroscopic image of subregion 7-1 through advance signal processing unit 17 processing and amplifying by computing machine 18.Second motor 12 drives platform 8 along the translation of Y direction by guide rail 10, and the next son zone 7-2 of biochip 7 is arrived within the visual field of flat-field objective 6.Repeat above-mentioned scanning process, obtain the fluoroscopic image of subregion 7-2,7-3,7-4 subregion.Complete biological chip fluorescent picture is obtained by each sub regions 7-1,7-2 of obtaining in computing machine 18 splicing flat-field objectives 6 visual fields, the fluoroscopic image of 7-3,7-4.

Claims (2)

1. the biochip imaging method of splitted with laser cofocus scanning combined image, step is as follows:
1) biochip is placed on the platform that moves by first motor and second driven by motor work two dimension;
2) laser that laser instrument is sent expands bundle through beam expanding lens, successively through arriving galvanometer after mirror reflects and the dichroscope transmission around the axle swing, make through the laser beam after the vibration mirror reflected and reflex to flat-field objective with different angles, flat-field objective is with the laser beam transverse focusing of the different incidence angles degree lower surface at biochip, to the biochip transversal scanning, simultaneously, the first driven by motor platform lengthwise movement, to the biochip longitudinal scanning, form the two-dimensional scan subregion of biochip;
3) fluorescence that sends of biochip is collected by flat-field objective, successively through galvanometer, dichroscope reflection, after the transmission of fluorescence color filter, arrive convergent mirror, on the focal plane of convergent mirror, place the confocal diaphragm that has aperture, make the fluorescence of biochip lower surface pass aperture and arrive photodetector, the output signal of photodetector is through advance signal processing unit processing and amplifying, by computer acquisition and rebuild the fluoroscopic image of this subregion;
4) the second driven by motor platform transverse translation is to biochip next son zone repeating step 2) and 3), obtaining each sub regions image, each the sub regions image by obtaining in the computing machine splicing flat-field objective visual field obtains complete biological chip fluorescent picture.
2. the device of biochip imaging method according to claim 1, it is characterized in that comprising laser instrument (1), beam expanding lens (2), catoptron (3), dichroscope (4), galvanometer (5), flat-field objective (6), fluorescence color filter (13), convergent mirror (14), have the confocal diaphragm (15) of aperture, photodetector (16) and be used to place the platform (8) of biochip (7), be provided with first guide rail (9) and second guide rail (10) in platform (8) bottom, the screw mandrel of first guide rail (9) links to each other with first motor (11) axle, the screw mandrel of second guide rail (10) links to each other with second motor (12) axle; Laser instrument (1), beam expanding lens (2), catoptron (3), dichroscope (4), galvanometer (5), flat-field objective (6) are arranged in order, form the laser scanning light path of biochip (7), galvanometer (5) is positioned at the front focal plane of flat-field objective (6), and biochip (7) is positioned at the back focal plane of flat-field objective (6); On the reflected light path of dichroscope (4), place color filter (13), convergent mirror (14), confocal diaphragm (15) and photodetector (16) successively along the light reflection direction, confocal diaphragm (15) is positioned at the back focal plane of convergent mirror (14).
CNB2005100970028A 2005-12-31 2005-12-31 Biochip imaging method splitted with laser cofocus scanning combined image and its device Expired - Fee Related CN100427926C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNB2005100970028A CN100427926C (en) 2005-12-31 2005-12-31 Biochip imaging method splitted with laser cofocus scanning combined image and its device

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB2005100970028A CN100427926C (en) 2005-12-31 2005-12-31 Biochip imaging method splitted with laser cofocus scanning combined image and its device

Publications (2)

Publication Number Publication Date
CN1793860A CN1793860A (en) 2006-06-28
CN100427926C true CN100427926C (en) 2008-10-22

Family

ID=36805429

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB2005100970028A Expired - Fee Related CN100427926C (en) 2005-12-31 2005-12-31 Biochip imaging method splitted with laser cofocus scanning combined image and its device

Country Status (1)

Country Link
CN (1) CN100427926C (en)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102944561B (en) * 2012-11-16 2015-07-29 北京金橙子科技有限公司 A kind of appearance detecting method of matrix form gadget and device
JP6463382B2 (en) * 2015-02-04 2019-01-30 ボッシュパッケージングテクノロジー株式会社 Inspection device and inspection system
CN106908407A (en) * 2017-02-22 2017-06-30 天津大学 A kind of pendular reflex scan-type multi-component material NDIR detection means
CN107254406B (en) * 2017-05-23 2019-07-05 北京大学 Biological cell chip high throughput, high intension, parallel imaging arrangement and screening system
CN109425597A (en) * 2017-09-04 2019-03-05 中国科学院上海光学精密机械研究所 The device and method of Sweat latent fingerprint detection on a kind of large format sample
CN109300116A (en) * 2018-09-03 2019-02-01 广东工业大学 The online defect identification method of laser welding based on machine learning
CN109342369A (en) * 2018-10-26 2019-02-15 中国科学院苏州生物医学工程技术研究所 The big visual field bio-imaging that is quickly detected for circulating tumor cell, scanning, analytical equipment
CN109580571B (en) * 2019-01-11 2021-05-04 中国科学院上海光学精密机械研究所 Detection device and detection method for potential fingerprints
CN112415733B (en) * 2020-12-11 2024-01-09 平湖莱顿光学仪器制造有限公司 Method, system, device and medium for controlling microscope to shoot sample image
CN113418932B (en) * 2021-06-30 2023-08-04 天津大学 Nondestructive inspection device and method for semiconductor wafer

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US549325A (en) * 1895-11-05 Suspender-buckle
US5381224A (en) * 1993-08-30 1995-01-10 A. E. Dixon Scanning laser imaging system
US5646411A (en) * 1996-02-01 1997-07-08 Molecular Dynamics, Inc. Fluorescence imaging system compatible with macro and micro scanning objectives
CN1385690A (en) * 2002-06-09 2002-12-18 浙江大学 Biochip analysis instrument

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US549325A (en) * 1895-11-05 Suspender-buckle
US5381224A (en) * 1993-08-30 1995-01-10 A. E. Dixon Scanning laser imaging system
US5646411A (en) * 1996-02-01 1997-07-08 Molecular Dynamics, Inc. Fluorescence imaging system compatible with macro and micro scanning objectives
CN1385690A (en) * 2002-06-09 2002-12-18 浙江大学 Biochip analysis instrument

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
光机二维扫描技术在激光共聚焦生物芯片扫描仪中的应用. 郑旭峰等.光学仪器,第25卷第4期. 2003
光机二维扫描技术在激光共聚焦生物芯片扫描仪中的应用. 郑旭峰等.光学仪器,第25卷第4期. 2003 *
双波长激光共聚焦生物芯片扫描仪的研制. 陆春光等.光学仪器,第27卷第5期. 2005
双波长激光共聚焦生物芯片扫描仪的研制. 陆春光等.光学仪器,第27卷第5期. 2005 *
激光共聚焦生物芯片扫描仪的研究. 王立强等.浙江大学学报(工学版),第39卷第4期. 2005
激光共聚焦生物芯片扫描仪的研究. 王立强等.浙江大学学报(工学版),第39卷第4期. 2005 *

Also Published As

Publication number Publication date
CN1793860A (en) 2006-06-28

Similar Documents

Publication Publication Date Title
CN100427926C (en) Biochip imaging method splitted with laser cofocus scanning combined image and its device
CN101101277B (en) High-resolution welding seam supersonic image-forming damage-free detection method
CN105911680B (en) SPIM microscopes with continuous sheet laser
CN102625022B (en) Rapid microscopic section scanning method with image space compensation motion and apparatus thereof
CN101050949A (en) Measuring system and its measuring method for large field object micro surface three dimension topography
EP1720003B1 (en) Multidirectional simultaneous observation optical system
CN102590155A (en) Tissue slice scanning and imaging device
JPH08252256A (en) Tomograph
CN111220615A (en) Inclined three-dimensional scanning microscopic imaging system and method
CN108956562A (en) A kind of light slice fluorescent microscopic imaging method and device based on reorientation
CN112817007A (en) Non-visual field scanning imaging system
CN107991766A (en) A kind of microscope and imaging method with three-dimensional imaging ability
JP2004317676A (en) Laser microscope, laser beam scanner, laser beam scanning method, and method for generating image data
JP2006275964A (en) Shading correction method for scanning fluorescence microscope
JP5848596B2 (en) Image acquisition device and focus method of image acquisition device
CN103299231B (en) Photo-scanning system
CN102466518A (en) Microscanning system and related method
EP1806575B1 (en) Examination apparatus
CN111103062B (en) Two-dimensional imaging device and method based on single photon counting
CN2551994Y (en) Scanner for biogene chip
CN1170183C (en) High-speed laser-confocal scanning microscopic imaging apparatus
JP5296861B2 (en) Image acquisition device and focus method of image acquisition device
CN107850766A (en) System and method for the image procossing in light microscope
CN214751082U (en) Laser scanning micro-measuring device
JP2003270545A (en) Optical scanning stage

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C17 Cessation of patent right
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20081022

Termination date: 20131231