CN1567559A - An orthogonal light path type fluorescent detection device for microchip analysis - Google Patents
An orthogonal light path type fluorescent detection device for microchip analysis Download PDFInfo
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- CN1567559A CN1567559A CN 03141572 CN03141572A CN1567559A CN 1567559 A CN1567559 A CN 1567559A CN 03141572 CN03141572 CN 03141572 CN 03141572 A CN03141572 A CN 03141572A CN 1567559 A CN1567559 A CN 1567559A
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Abstract
The invention is an orthogonal light path type fluorescent detecting device used in microanalytical chip, composed of a light source, an optical system, and a detecting system, and its characteristic lies in that the optical system adopts an orthogonal light path, the laser emitted from the light source reaches the microanalytical chip from the side wall, and the detecting system detects the fluorescent light outgoing from the side wall. The advantages: the optical system is simple, the device is easy to make, the making cost is low, the detecting sensitivity is high, and the system is easily integrated and miniaturized.
Description
Technical field
The present invention relates to the micro-fluidic chip analysis, particularly relate to a kind of quadrature light path type fluorescence detection device of microanalysis chip.
Background technology
The field that this patent relates to is the research of the detection system on the relevant microanalysis chip.Detection system is the important component part of an analytical system.Microanalysis chip is little because of its chip volume, and sample size only is skin, the upgrading of receiving, and the reaction channel of detection place is generally tens microns wide, and therefore the requirement to its detection means and device has its particularity.
One of main detection method of using in the microanalysis chip is a fluorescence method at present, wherein use laser-induced fluorescence (LIF) (the Laser Induced Fluorescence of laser as excitation source, LIF) detection method is highly sensitive because of having, and more easily is implemented in the characteristics of the detection on the chip and becomes the method that widely adopts at present.At present, the optical system of fluorescence detection device mainly contains two kinds: a kind of is oblique fire type light path system, exciting light is from angle at 45, chip plane and perpendicular to detection zone in the direction irradiation tunnel of microchannel, with incident light angle at 45 and the direction vertical with the chip plane on detect fluorescence signal with photoelectric detector, advantage is that light path is simple, shortcoming is that exciting light is stronger to the interference of fluorescence, and detection sensitivity is low; Another kind is confocal type light path, mainly is made of dichroscope, collector lens, interferometric filter etc.Laser beam is through beam-expanding collimation, focus on the surveyed area of back vertical irradiation by dichroic mirror reflects and by microcobjective to chip, the fluorescence that excites generation is through the dichroscope transmission, and by the micro-eyepiece focusing coaxial and confocal with microcobjective, imaging detects on detector behind the further filtering interfering light of interferometric filter again, wherein dichroscope is crucial optical element, plays the effective centrifugation to exciting light and fluorescence.Altogether the advantage of focus type fluorescence detector is that detection sensitivity is very high, but such optical system structure complexity costs an arm and a leg, and is difficult for microminiaturized, integrated.
Summary of the invention
The objective of the invention is to design that a kind of volume is little, optical texture is simple, detection sensitivity is high, be easy to microanalysis chip fluorescence detection device integrated and with low cost.
The present invention mainly is made of light source, optics and detection system three parts.Light source can adopt laser, continuous light source, light-emitting diode etc.Wherein optical system adopts the quadrature detection light path, and the exciting light of light emitted is from the sidewall incident of microanalysis chip, and detection system detects the fluorescence via the outgoing of chip sidewall.Purpose is to simplify light path system, reduces the interference of exciting light to fluoroscopic examination, improves phosphor collection efficient.
According to the present invention, the microanalysis chip that uses is characterized in that: (a) chip at least by last (1), down (2) two form, in last (1) or sheet (2) processing microchannel down,, form the microchannel of sealing with chip (1) and chip (2) involution; (b) chip is long is the 0.5-300 millimeter, and wide is the 0.5-300 millimeter, and thickness is the 0.1-50 millimeter; (c) microchannel width 1-10000 micron, degree of depth 1-5000 micron.
According to the present invention, light path system is divided into two big classes: the exciting light of (1) light emitted is from the sidewall incident of microanalysis chip; (2) detection system detects the fluorescence via the outgoing of chip sidewall.
According to the present invention, first kind light path system is characterized in that, exciting light detects by fluorescence hot spot emitted fluorescence on other direction that departs from exciting light light path direction via microchannel in the chip sidewall incident chip.Usually detect fluorescence above or below chip, this kind mode helps simplifying the setting of detection system.Because the scattering of exciting light in the microchannel is different in different directions, therefore, by selecting the angle between suitable exciting light and the detected fluorescence, be to carry out fluoroscopic examination on the more weak direction of selective exitation light scattering intensity, the scattered light that can effectively reduce exciting light and produced is to the interference of fluoroscopic examination.The typical structure of this type of light path system is: exciting light via microchannel in the chip sidewall incident chip, detects fluorescence along becoming 90 ° of angle directions with the microchannel on the direction that becomes 90 ° of angles with the exciting light direction.
According to the present invention, the second class light path system is characterized in that, exciting light is via chip plane top, perhaps below, and perhaps microchannel in the sidewall incident chip, detection system detects by the emission of fluorescence hot spot, via the fluorescence of chip sidewall outgoing.This kind mode helps simplifying the setting of detection system, simultaneously, detects fluorescence by the chip sidewall, and the interference that the exciting light scattering causes is less.For fluorescence by the outgoing of chip sidewall, detect fluorescence by selecting suitable fluorescence exit direction and the angle between the microchannel, the scattered light that can further reduce exciting light and produced is to the influence of fluoroscopic examination.The typical structure of this type of light path system is: exciting light is along becoming microchannel in 90 ° of angle direction vertical incidence chips with the chip plane, detection system detects edge and the fluorescence of microchannel angle at 45 via the outgoing of chip sidewall.
According to the present invention, for employed microanalysis chip, for reducing the light loss that scattering caused by the chip sidewall time of exciting light or fluorescence, needing has the zone of exciting light or detected fluorescence process to carry out the processing that surface finish or additional coatings etc. can increase the light transmission amount, reduce light scattering to the chip sidewall.
According to the present invention,, it is characterized in that should trying one's best near the chip side mural margin in the detected zone in the microchannel for employed microanalysis chip, shorten fluorescence by the light path of shot point to detection window, help improving the collection efficiency of fluorescence.
Major advantage of the present invention is:
Optical system structure is simple, is easy to build, and the detection system volume is little, helps realizing the microminiaturization of microanalysis chip system and integrated.Sidewall by utilizing chip is as the incidence zone of exciting light or detect the outgoing district of fluorescence, has reduced the interference of exciting light to fluorescence effectively, and detection system can obtain higher detection sensitivity.System cost is cheap, helps extensively popularizing of this technology.
Following with reference to the present invention of accompanying drawing detailed description.
Description of drawings
Fig. 1 is 1 a end view according to a preferred embodiment of the present invention.
Fig. 2 is 1 a vertical view according to a preferred embodiment of the invention.
Fig. 3 is 2 a end view according to a further advantageous embodiment of the invention.
Fig. 4 is 1 microanalysis chip channel configurations figure according to a preferred embodiment of the invention.
Fig. 5 is another microanalysis chip channel configurations figure of 1 according to a preferred embodiment of the invention
Fig. 6 is that 1 device detects the outcome record figure of fluorescence according to a preferred embodiment of the invention.
Fig. 7 is that 2 device detects the outcome record figure of fluorescence according to a preferred embodiment of the invention.
Embodiment
With reference to accompanying drawing, below will describe in detail according to a preferred embodiment of the invention.
With reference to accompanying drawing 1, microchip is made up of last 1, following 2 liang of sheet glass, and sheet glass thickness is 1.7 millimeters.Adopt photomask and wet etching technique at last 1 processing channel, microchannel 6 width are 60 microns, 20 microns of the degree of depth.Adopt the method for classical high temperature bonding to realize last 1, following 2 two permanent involution.Obtaining length is 60 millimeters, the micro-fluidic chip that width is 15 millimeters.In the present embodiment, excitation source 7 is the semiconductor pump laser of 473nm; The exciting light 5 that laser sends is after collecting objective 4 focuses on, from the direction incident chip microchannel 6 perpendicular to 2 planes of sheet under the chip; Selection is by fluorescence hot spot 11 emission, from the fluorescence 8 of chip sidewall 3 outgoing, collected by the collection object lens 9 that are positioned at behind the detection window, and that gathers object lens is provided with direction and chip sidewall 3 angles at 45; The fluorescence that object lens are collected mating plate after filtration filters, and detects fluorescence signal by photoelectric detector 10--photomultiplier at last.Photoelectric detector links to each other with extraneous data recordin and procesin system by data wire.
Fig. 2 is 1 a vertical view according to a preferred embodiment of the invention.Fluorescence hot spot 11 in the chip microchannel 6 to around emitting fluorescence 8.Be chosen on the direction with chip sidewall 3 angles at 45 and collect fluorescence, fluorescence is collected by gathering lens 9, mating plate light after filtration again, and last fluorescence signal intensity is detected by photomultiplier 10.
With reference to accompanying drawing 3, after the exciting light 5 that is sent by light source 7 is assembled through focusing objective lens 4, from the sidewall 3 of chip in direction incident perpendicular to chip microchannel 6; Selection is collected through gathering object lens 9, and by filter filtering stray light, is detected fluorescence signals by photoelectric detector 10 at last from the fluorescence 8 perpendicular to the direction outgoing of sheet under the chip 2.
Fig. 4 is 1 microanalysis chip channel configurations figure according to a preferred embodiment of the invention.The split tunnel of the microchannel 6 on the chip is partly through the bend design, and surveyed area 12 is near near chip sidewall 3 edges endways, and purpose shortens fluorescence by the distance of hot spot to detection window, improves the collection efficiency of fluorescence.
Fig. 5 is another microanalysis chip channel configurations figure of 1 according to a preferred embodiment of the invention.The split tunnel part of the microchannel 6 on the chip comprises its end point detection zone 12, and near near chip sidewall 3 edges, purpose is to shorten fluorescence by the light path of shot point to detection window, helps improving the collection efficiency of fluorescence.
Fig. 6 is that 1 device detects the outcome record figure of fluorescence according to a preferred embodiment of the invention.The fluorometric reagent that adopts is the fluorescein sodium salting liquid, and concentration is 10
-6Mol/L; Buffer solution is the borate buffer solution of 5mmol/L (pH=9.2); Excitation source is the semiconductor pump laser of 473nm, detects fluorescence signal with photomultiplier.Blank solution signal strength signal intensity 30mV, fluorescence signal intensity 110mV.
Fig. 7 is that 2 device detects the outcome record figure of fluorescence according to a preferred embodiment of the invention.The fluorometric reagent that adopts is the fluorescein sodium salting liquid, and concentration is 10
-6Mol/L; Buffer solution is the borate buffer solution of 5mmol/L (pH=9.2); Excitation source is the semiconductor pump laser of 473nm, detects fluorescence signal with photomultiplier.The blank solution signal strength signal intensity about 25mV, fluorescence signal intensity 40mV.
Claims (8)
1. quadrature light path type fluorescence detection device that is used for microanalysis chip, this device is by light source, optical system, detection system three parts constitute, it is characterized in that described optical system adopts the quadrature detection light path, the exciting light of light emitted is from the sidewall incident of microanalysis chip, detection system detects on other direction that departs from exciting light light path direction by fluorescence hot spot emitted fluorescence, perhaps the exciting light of light emitted is from the top, plane of microanalysis chip, perhaps, perhaps sidewall incident, the detection system detection is launched by the fluorescence hot spot, via the fluorescence of chip sidewall outgoing.
2. according to the described quadrature light path type fluorescence detection device that is used for microanalysis chip of claim 1, it is characterized in that, described microanalysis chip at least by last (1), down (2) two form, in last (1) or following sheet (2) processing microchannel, with chip (1) and chip (2) involution, form the microchannel (6) of sealing; Chip is long to be the 0.5-300 millimeter, and wide is the 0.5-300 millimeter, and thickness is the 0.1-50 millimeter; Microchannel width 1-10000 micron, degree of depth 1-5000 micron.
3. the quadrature light path type fluorescence detection device that is used for microanalysis chip according to claim 1, it is characterized in that, exciting light (5) is by microchannel (6) in chip sidewall (3) the incident chip, and detector (10) detects by fluorescence hot spot (11) emitted fluorescence (8) departing from other direction of exciting light light path.
4. the quadrature light path type fluorescence detection device that is used for microanalysis chip according to claim 3, it is characterized in that, exciting light (5) becomes 90 ° of angle directions with microchannel (6), via microchannel (6) in chip sidewall (3) the incident chip, on the direction that becomes 90 ° of angles with the exciting light direction, detect fluorescence.
5. the quadrature light path type fluorescence detection device that is used for microanalysis chip according to claim 1, it is characterized in that, exciting light (5) is via top, chip plane (1), or below (2), or sidewall incident chip microchannel (6), the detection system detection is launched by fluorescence hot spot (11), via the fluorescence (8) of chip sidewall outgoing, select suitable fluorescence exit direction and the angle between the microchannel to detect fluorescence, the scattered light that can effectively reduce exciting light and produced is to the influence of fluoroscopic examination.
6. the quadrature light path type fluorescence detection device that is used for microanalysis chip according to claim 5, it is characterized in that, exciting light (5) becomes 90 ° of angle direction incident chip microchannels (6) with the chip plane, detection system detects edge and microchannel (6) angle at 45, detects the fluorescence via chip sidewall (3) outgoing.
7. the quadrature light path type fluorescence detection device that is used for microanalysis chip according to claim 2, it is characterized in that microanalysis chip sidewall (3) has exciting light or has the zone of detected fluorescence process to carry out surface finish or additional coatings, can increase the light transmission amount, reduce the processing of light scattering.
8. the quadrature light path type fluorescence detection device that is used for microanalysis chip according to claim 2, it is characterized in that detected zone (12) in the microanalysis chip microchannel (6) should try one's best near chip sidewall (3) edge, help improving the collection efficiency of fluorescence.
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| CN 03141572 CN1285902C (en) | 2003-07-08 | 2003-07-08 | An orthogonal light path type fluorescent detection device for microchip analysis |
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| CN 03141572 CN1285902C (en) | 2003-07-08 | 2003-07-08 | An orthogonal light path type fluorescent detection device for microchip analysis |
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| CN1285902C CN1285902C (en) | 2006-11-22 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101949847A (en) * | 2010-08-13 | 2011-01-19 | 浙江大学 | Lensless fluorescence imaging detection device and application |
| CN102105800A (en) * | 2008-07-22 | 2011-06-22 | 爱科来株式会社 | Microchip and analyzing apparatus |
| CN104198452A (en) * | 2014-09-12 | 2014-12-10 | 四川大学 | Signal enhancement laser-induced fluorescence system |
-
2003
- 2003-07-08 CN CN 03141572 patent/CN1285902C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102105800A (en) * | 2008-07-22 | 2011-06-22 | 爱科来株式会社 | Microchip and analyzing apparatus |
| US8436990B2 (en) | 2008-07-22 | 2013-05-07 | Arkray, Inc. | Microchip and analyzing apparatus |
| CN101949847A (en) * | 2010-08-13 | 2011-01-19 | 浙江大学 | Lensless fluorescence imaging detection device and application |
| CN101949847B (en) * | 2010-08-13 | 2012-05-30 | 浙江大学 | Lensless fluorescence imaging detection device and application |
| CN104198452A (en) * | 2014-09-12 | 2014-12-10 | 四川大学 | Signal enhancement laser-induced fluorescence system |
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| CN1285902C (en) | 2006-11-22 |
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