CN101464411A - Capillary array analyzer - Google Patents
Capillary array analyzer Download PDFInfo
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- CN101464411A CN101464411A CNA200910095512XA CN200910095512A CN101464411A CN 101464411 A CN101464411 A CN 101464411A CN A200910095512X A CNA200910095512X A CN A200910095512XA CN 200910095512 A CN200910095512 A CN 200910095512A CN 101464411 A CN101464411 A CN 101464411A
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- 238000001499 laser induced fluorescence spectroscopy Methods 0.000 description 4
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- 238000012742 biochemical analysis Methods 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
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Abstract
The invention belongs to the technical field of bioanalysis, and relates to a capillary array analyzer, wherein, exciting light sent out by a light source is reflected to a vibrating mirror by a reflecting mirror and a fluorescence separating mirror; as the vibrating swings around a rotating shaft in a reciprocating manner, the exciting light also swings in a reciprocating manner; then the exciting light arrives at a scanning objective; the exciting light is focused on a focal plane for accommodating a capillary array by the scanning objective and is converted into one-dimensional translational scanning on the focal plane by the scanning objective; during the scanning process, samples marked with fluorescent substances in capillaries are excited by the exciting light in sequence; fluorescence emitted by the fluorescent substances marked on the samples is collected by the scanning objective and converted into a parallel beam that is coaxial with the exciting light and spreads in the opposite direction; the parallel beam is separated from the exciting light through reflection and then arrives at a convergent lens; and the parallel beam of fluorescence, from which the background light is filtered by a fluorescence color filter on a color wheel, is detected by a photodetector after passing through a confocal ostiole. The capillary array analyzer provided by the invention has the advantages of simple structure, small size, low cost and excellent performance.
Description
Technical field
The invention belongs to the bioassay technique field, particularly a kind of capillary array analyzer.
Background technology
Capillary Electrophoresis is to pass through effect of electric field, sample is moved in kapillary, and different compositions has different migration velocities in the sample, thereby realize separating, and the detection window place in kapillary surveys, and by the detectable signal of gained sample analyzed at last.This technology is widely used in biochemical analysis; as DNA, Separation of Proteins analysis; its have separation efficiency height, sample consumption less, the automaticity height, it has a wide range of applications in dna sequencing, biological and fields such as clinical research, legal medical expert's evaluation, environmental protection, analytical chemistry and food inspection.The detection of electrophoresis sample mainly contains isotope labeling, chemiluminescence method, surface plasma resonance and laser-induced fluorescence (LIF), adopts optical detection usually, and its medium sensitivity is the highest, and most widely used be laser-Induced Fluorescence Detection.
In order to realize high throughput analysis, can be with many capillaries while electrophoresis, detection window is lined up array, carries out parallel detection, is generally the capillary array electrophoresis fluoroscopic examination.Present capillary array electrophoresis detection method mainly contains following several: (1) is arranged in parallel kapillary, be imaged onto by image-forming objective lens and carry out non-scanning non-confocal detection on the CCD, but the detection sensitivity of non-confocal mode is restricted.(2) kapillary is arranged in parallel, and places on the scanning platform, the motion realization scanning confocal detection by scanning platform perhaps by the relative capillary motion of camera lens, realizes the scanning to capillary array.The sweep velocity of this scan mode is limited by driver, and vibrations are big.(3) with capillary array on a cylinder, camera lens perpendicular to the central shaft arrangement of cylinder, moves in a circle around central shaft by camera lens and to realize scanning in cylinder.This mode needs kapillary by circumferential arrangement, thereby makes troubles to some analyses and application.(4) kapillary or microchannel radial arrays are arranged on the disk plane, camera lens disposes perpendicular to disk, and rotates around disc centre, realizes scanning.But disk capillaceous is arranged also can give and is analyzed and use and make troubles, and rotation of lens mechanism complexity, is difficult for miniaturization.
At present in the confocal detection that adopts, the multicolor fluorescence detection technique adopts color filter beam split mode, adopt a color filter earlier the fluorescence of four wavelength period to be separated into two light paths by long wave and shortwave as 4 look fluoroscopic examinations, each light path is separated into two light paths by color filter more then, and each light path adopts a photo-detector to survey.The photo-detector that this beam split detection mode at first needs is many, has increased the volume of production cost and detector; Secondly, because of color filter is many, the fluorescence loss is many, has therefore limited detection sensitivity; At last, because many color filters are configured in the light path, therefore, space requirement is big, has further increased the volume of detector.
Summary of the invention
The invention provides a kind of simple in structure, volume is little, cost is low and the capillary array analyzer of function admirable.
The technical solution used in the present invention is as follows: capillary array analyzer, comprise light source, catoptron, galvanometer, scanning objective, fluorescence splitting mirror, convergent lens, colour wheel, confocal aperture, photo-detector, the exciting light that light source sends is by mirror reflects, see through the fluorescence splitting mirror and incide galvanometer, galvanometer is reciprocating swing around the shaft, and makes also reciprocally swinging of exciting light; Back exciting light incides scanning objective, scanning objective is poly-to the focal plane of placing capillary array with exciting light, the swing of exciting light scans through the one-dimensional translation that scanning objective is converted on the focal plane, in scanning process, exciting light excites the sample that is marked with fluorescent material in the kapillary successively, the fluorescence that fluorescent material sent of sample mark is collected by scanning objective, and be converted to the fluorescence parallel beam of and backpropagation coaxial with exciting light, the fluorescence parallel beam is reflected and is separated with excitation beam at fluorescence splitting mirror place, after incide convergent lens, fluorescence parallel beam after the convergence is through the fluorescence color filter wiping out background light on the colour wheel, focus on by confocal aperture, survey by photo-detector.
Described capillary array analyzer, the kapillary of capillary array are parallel to a plane, or capillary array is the micro channel array that is etched in the chip.
Described capillary array analyzer, capillary array is sandwiched between two substrates.
Described capillary array analyzer, light source adopts laser or light emitting diode.
Described capillary array analyzer, light source, capillary array, confocal aperture three are in conjugate position, form confocal detection.
Described capillary array analyzer, scanning objective adopt picture side's heart f-theta object lens far away.
Described capillary array analyzer, the optical axis of fluorescence splitting mirror and exciting light is at 45.
Described capillary array analyzer, the fluorescence splitting mirror adopts dichroic mirror or center total reflective mirror with holes.
Described capillary array analyzer, photo-detector adopts photomultiplier, or Avalanche Photo Diode, or the intrinsic photodiode.
Described capillary array analyzer, the fluorescence color filter is mounted in the color filter group on the colour wheel, and colour wheel rotates the color filter of different spectral characteristics is in the light path, surveys the fluorescence of different wave length.
In the course of work, buffer pool is inserted at the kapillary two ends, and is connected with the both positive and negative polarity of power supply respectively, charge into sieving media in the kapillary, the both positive and negative polarity making alive carries out prerunning to kapillary, treat current stabilization after, charged character per sample is at the negative or positive electrode sample introduction.Behind the sample introduction, sample in the kapillary is carried out electrophoretic separation, when the sample that has a fluorescent material is moved to detection window, being scanned this hot spot that excites capillaceous excites, the fluorescence that sends is collected through scanning objective, and galvanometer and the reflection of fluorescence splitting mirror are assembled on the convergence object lens, fluorescence color filter colour filter by on the colour wheel is detected by photo-detector.
To the homogeneity of every capillary, scanning objective adopts heart f-theta object lens far away, makes the chief ray of its each scanning position all be parallel to optical axis when guaranteeing scanning, and system do not have vignetting, thereby guarantees that each scanning position collects the consistance of fluorescence.The f-theta camera lens make vibration mirror reflected in the scanning process to the angle change transitions of scanning objective light beam be capillary array linear displacement on the whole change, guarantee the homogeneity of sweep velocity.Adopt the mode of galvanometer among the present invention, have advantages such as scanning inertia is little, moving component is few, detector stable performance, detection homogeneity height, compact conformation in conjunction with the capillary array laser-Induced Fluorescence Detection of heart f-theta object lens far away.
The present invention adopts the colour wheel rotation, makes a plurality of color filter time division multiplexes ground carry out multicolor fluorescence and detects, and has reduced the loss of fluorescent energy, has reduced the number of the photo-detector that uses, thereby has both reduced manufacturing cost, reduces volume again.
Description of drawings
Fig. 1 is a structural representation of the present invention.
Fig. 2 is that capillary array is arranged synoptic diagram.
Fig. 3 is a colour wheel rotary multi-color fluoroscopic examination synoptic diagram.
Embodiment
Below in conjunction with accompanying drawing the present invention is elaborated.
Fig. 1 is a structural representation of the present invention.Light source 1 adopts laser, and the emergent light direction of light source 1 is installed catoptron 3, and the exciting light 2 that light source 1 sends is by catoptron 3 reflections, and this catoptron 3 can be used for the light path of exciting light 2 to be regulated.The emergent light direction of catoptron 3 is installed fluorescence splitting mirror 4, the optical axis angle at 45 of fluorescence splitting mirror 4 and exciting light 2.Fluorescence splitting mirror 4 is the total reflective mirrors that the center is porose, and center pit is used to pass exciting light 2, and total reflective mirror is used for reflected fluorescent light.The center pit that sees through fluorescence splitting mirror 4 by the exciting light after the reflection 2 incides galvanometer 5, and galvanometer 5 is installed in the rotating shaft 501, can be around the axle reciprocally swinging.The galvanometer eyeglass also is a total reflective mirror, exciting light 2 is reflected by galvanometer 5, along with galvanometer 5 501 swing around the shaft, exciting light 2 after the reflection is also in swing, the exciting light 2 of swing reenters and is mapped to scanning objective 6, scanning objective 6 adopts picture side's heart f-theta object lens far away, and scanning objective 6 converges to exciting light on the focal plane of these object lens.The capillary array of being made up of kapillary 7 places on the focal plane of scanning objective 6, and the swing of exciting light 2 scans through the one-dimensional translation that scanning objective 6 is transformed on the focal plane.In the scanning process, exciting light 2 sequentially excites the sample that is marked with fluorescent material in each kapillary, the fluorescence that fluorescent material sent 8 of mark sample is collected by scanning objective 6, and be transformed to the directional light of and backpropagation coaxial with exciting light, fluorescence parallel beam 8 is reflected at fluorescence splitting mirror 4 places and separates with excitation beam 2, incide convergent lens 9 then, fluorescence after the convergence is through the fluorescence color filter 101 wiping out background light on the colour wheel 10, focus on by confocal aperture 11, surveyed by photo-detector 12, photo-detector 12 adopts photomultiplier.Confocal aperture 11 threes before light source 1, capillary array 7, the photo-detector 12 are in conjugate position, form confocal detection.
Fig. 2 is the arrangement synoptic diagram of capillary array 7, many capillaries 71 are arranged in parallel in one plane, these kapillaries are positioned to be carved with the kapillary fixing base 72 of V-type groove, there is a hole to be used for seeing through exciting light and fluorescence in the middle of the substrate 72, substrate 73 is fixing with substrate 72 by screw 74, kapillary 71 is fixed on two substrates, 72,73 centres, and substrate 73 is the plane with the surface of contact of kapillary 7.
Fig. 3 is the synoptic diagram that colour wheel rotation time division multiplex detects multicolor fluorescence, on the colour wheel 10 fixedly polylith see through the different color filter 101 of spectrum, the colour filter face of color filter 101 is vertical with the central shaft of fluorescence beam 8, and the rotating shaft center 102 of colour wheel and the misalignment of fluorescence beam 8, make in the process of colour wheel 10 rotations, the central shaft of each color filter 101 just in time can overlap with the central shaft of fluorescence beam 8.By rotation colour wheel 10, during 101 pairs of fluorescence beam 8 colour filters of each color filter, it is that the fluorescence of different colours sees through color filter 101 and arrives photo-detectors 12 that different spectral components are arranged, and realizes time-multiplexed multicolor fluorescence acquisition of signal.
Certainly; those of ordinary skill in the art should be familiar with; above-mentioned just explaining to a specific embodiment of the present invention; and be not limitation of the invention; can adopt light emitting diode as light source; capillary array can be the micro channel array that is etched in the chip; the fluorescence splitting mirror can adopt dichroic mirror; photo-detector can adopt Avalanche Photo Diode or intrinsic photodiode or the like; every in essential scope of the present invention, all will fall within protection scope of the present invention variation, the modification of above embodiment.
Claims (10)
1, capillary array analyzer, comprise light source, catoptron, galvanometer, scanning objective, it is characterized in that: also comprise fluorescence splitting mirror, convergent lens, colour wheel, confocal aperture, photo-detector, the exciting light that light source sends is by mirror reflects, see through the fluorescence splitting mirror and incide galvanometer, galvanometer is reciprocating swing around the shaft, and makes also reciprocally swinging of exciting light; Back exciting light incides scanning objective, scanning objective is poly-to the focal plane of placing capillary array with exciting light, the swing of exciting light scans through the one-dimensional translation that scanning objective is converted on the focal plane, in scanning process, exciting light excites the sample that is marked with fluorescent material in the kapillary successively, the fluorescence that fluorescent material sent of sample mark is collected by scanning objective, and be converted to the fluorescence parallel beam of and backpropagation coaxial with exciting light, the fluorescence parallel beam is reflected and is separated with excitation beam at fluorescence splitting mirror place, after incide convergent lens, fluorescence parallel beam after the convergence is through the fluorescence color filter wiping out background light on the colour wheel, focus on by confocal aperture, survey by photo-detector.
2, by the described capillary array analyzer of claim 1, it is characterized in that: the kapillary of described capillary array is parallel to a plane, or capillary array is the micro channel array that is etched in the chip.
3, by the described capillary array analyzer of claim 2, it is characterized in that: described capillary array is sandwiched between two substrates.
4, by the described capillary array analyzer of claim 1, it is characterized in that: described light source adopts laser or light emitting diode.
5, by each described capillary array analyzer of claim 1-4, it is characterized in that: light source, capillary array, confocal aperture three are in conjugate position, form confocal detection.
6, by the described capillary array analyzer of claim 1, it is characterized in that: described scanning objective adopts picture side's heart f-theta object lens far away.
7, by the described capillary array analyzer of claim 1, it is characterized in that: the optical axis of described fluorescence splitting mirror and exciting light is at 45.
8, by the described capillary array analyzer of claim 7, it is characterized in that: described fluorescence splitting mirror adopts dichroic mirror or center total reflective mirror with holes.
9, by the described capillary array analyzer of claim 1, it is characterized in that: described photo-detector adopts photomultiplier, or Avalanche Photo Diode, or the intrinsic photodiode.
10, by the described capillary array analyzer of claim 1, it is characterized in that: described fluorescence color filter is mounted in the color filter group on the colour wheel, and colour wheel rotates the color filter of different spectral characteristics is in the light path, surveys the fluorescence of different wave length.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200910095512XA CN101464411A (en) | 2009-01-15 | 2009-01-15 | Capillary array analyzer |
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| Application Number | Priority Date | Filing Date | Title |
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| CNA200910095512XA CN101464411A (en) | 2009-01-15 | 2009-01-15 | Capillary array analyzer |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063646A (en) * | 2013-01-14 | 2013-04-24 | 武汉大学 | Dual-purpose laser-induced fluorescence detecting system |
| CN107741411A (en) * | 2017-10-13 | 2018-02-27 | 中国科学院上海技术物理研究所 | A kind of isometric helix multiple spot ultraviolet spectra freezes serum analysis instrument |
| CN107907523A (en) * | 2017-10-13 | 2018-04-13 | 中国科学院上海技术物理研究所 | A kind of perseverance low temperature isometric helix multiple spot ultraviolet spectra freezes serum analysis method |
| WO2018082136A1 (en) * | 2016-11-07 | 2018-05-11 | 中国科学院光电研究院 | Scanning type laser-induced spectral plane range analysis and detection system |
| CN108362258A (en) * | 2018-02-09 | 2018-08-03 | 滁州职业技术学院 | A kind of rotary measurement device applied to grooving processing |
| CN109813692A (en) * | 2019-01-02 | 2019-05-28 | 北京科技大学 | A kind of capillary analysis detection method based on ultrasound aggregation |
| US20210341420A1 (en) * | 2020-04-30 | 2021-11-04 | Promega Corporation | Laser illumination techniques for capillary electrophoresis |
| CN114002196A (en) * | 2016-02-22 | 2022-02-01 | 株式会社日立高新技术 | Luminescence detection device |
| GB2599049A (en) * | 2017-02-20 | 2022-03-23 | Hitachi High Tech Corp | Analysis system and analysis method |
| CN114317698A (en) * | 2021-12-17 | 2022-04-12 | 青岛松辉光电有限公司 | Reflective PCR instrument |
| CN115747035A (en) * | 2023-01-05 | 2023-03-07 | 苏州和迈精密仪器有限公司 | Continuous scanning monitoring analysis system based on discrete three-dimensional fluorescence technology |
| US11860094B2 (en) | 2017-02-20 | 2024-01-02 | Hitachi High-Tech Corporation | Analysis system and analysis method |
-
2009
- 2009-01-15 CN CNA200910095512XA patent/CN101464411A/en active Pending
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063646B (en) * | 2013-01-14 | 2015-04-29 | 武汉大学 | Dual-purpose laser-induced fluorescence detecting system |
| CN103063646A (en) * | 2013-01-14 | 2013-04-24 | 武汉大学 | Dual-purpose laser-induced fluorescence detecting system |
| CN114002196A (en) * | 2016-02-22 | 2022-02-01 | 株式会社日立高新技术 | Luminescence detection device |
| US10823679B2 (en) | 2016-11-07 | 2020-11-03 | Academy Of Opto-Electronics Chinese Academy Of Sciences | Scanning type laser induced spectrum analysis and detection system |
| WO2018082136A1 (en) * | 2016-11-07 | 2018-05-11 | 中国科学院光电研究院 | Scanning type laser-induced spectral plane range analysis and detection system |
| GB2599049B (en) * | 2017-02-20 | 2022-11-09 | Hitachi High Tech Corp | Capillary electrophoresis device |
| GB2599049A (en) * | 2017-02-20 | 2022-03-23 | Hitachi High Tech Corp | Analysis system and analysis method |
| US11860094B2 (en) | 2017-02-20 | 2024-01-02 | Hitachi High-Tech Corporation | Analysis system and analysis method |
| CN107907523A (en) * | 2017-10-13 | 2018-04-13 | 中国科学院上海技术物理研究所 | A kind of perseverance low temperature isometric helix multiple spot ultraviolet spectra freezes serum analysis method |
| CN107741411A (en) * | 2017-10-13 | 2018-02-27 | 中国科学院上海技术物理研究所 | A kind of isometric helix multiple spot ultraviolet spectra freezes serum analysis instrument |
| CN108362258A (en) * | 2018-02-09 | 2018-08-03 | 滁州职业技术学院 | A kind of rotary measurement device applied to grooving processing |
| CN109813692A (en) * | 2019-01-02 | 2019-05-28 | 北京科技大学 | A kind of capillary analysis detection method based on ultrasound aggregation |
| US20210341420A1 (en) * | 2020-04-30 | 2021-11-04 | Promega Corporation | Laser illumination techniques for capillary electrophoresis |
| US12019046B2 (en) * | 2020-04-30 | 2024-06-25 | Promega Corporation | Laser illumination techniques for capillary electrophoresis |
| CN114317698A (en) * | 2021-12-17 | 2022-04-12 | 青岛松辉光电有限公司 | Reflective PCR instrument |
| CN115747035A (en) * | 2023-01-05 | 2023-03-07 | 苏州和迈精密仪器有限公司 | Continuous scanning monitoring analysis system based on discrete three-dimensional fluorescence technology |
| US11988632B1 (en) | 2023-01-05 | 2024-05-21 | Suzhou Helmen Precision Instruments Co., Ltd. | Systems for continuous scanning monitoring and analysis based on discrete three-dimensional fluorescence technology |
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