CN101153869A - Device for high-throughout monitoring micro-array biomolecule reaction by light reflection difference method - Google Patents
Device for high-throughout monitoring micro-array biomolecule reaction by light reflection difference method Download PDFInfo
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- CN101153869A CN101153869A CNA2006101135764A CN200610113576A CN101153869A CN 101153869 A CN101153869 A CN 101153869A CN A2006101135764 A CNA2006101135764 A CN A2006101135764A CN 200610113576 A CN200610113576 A CN 200610113576A CN 101153869 A CN101153869 A CN 101153869A
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Abstract
The invention provides a device for high-flux monitoring of micro array biomolecular reaction through light reflex difference method, comprising an incident light path, a sample table, an emergent light path and a data acquisition and processing system, wherein, the incident light path comprises a laser, a polarizer, a photoelastic modulator, a phase shifter and a beam expander arranged between the phase shifter and the sample table; the emergent light path comprises a polarization analyzer and an electrooptical signal converter; the electrooptical signal converter comprises a silicon photodiode array, a pulse switching circuit, a first phase-locked amplifier and a second phase-locked amplifier; the output end of the silicon photodiode array is respectively in electric connection with the input ends of the pulse switching circuit, the first phase-locked amplifier and the second phase-locked amplifier. With a simple structure and easy operation, the device reduces the noise caused by moving a sample and greatly improves signal-noise ratio; meanwhile, the device can complete non-mark quick high-flux monitoring of the reactions of the array elements of a micro array at the same time, thereby greatly increasing monitoring efficiency.
Description
Technical field
The present invention relates to a kind of monitoring device of micro-array biomolecule reaction, the device of the quick no-mark monitoring biomolecular reaction of particularly a kind of high flux.
Background technology
The method that the monitoring bio molecule is commonly used is fluorescent marker method and surface plasma resonance method.Fluorescence labeling is a kind of monitoring method that diminishes commonly used, and there is the observation process complexity in this method, can changes the structure of protein molecule and produces shortcomings such as photobleaching phenomenon; Surface plasma resonance is the method for the present no-mark monitoring bio-molecular interaction of using always, and this kind method requires biomolecule microarry to constitute on functional gold thin film, and very high to the thickness and the uniformity requirement of gold thin film.
Oblique incident ray difference in reflection technology (Oblique-incidence reflectivity difference, hereinafter to be referred as OIRD) be a kind of optical measuring technique of highly sensitive special surface measurements Film Optics character, it can monitor relative reflectance Δ R/R=1 * 10 in the thin film growth process
-5With the variation of cone of coverage Δ θ=0.02, be widely used in the noninvasive monitoring of real-time in-situ of the growth course of membraneous material always.This method also is applied in the biomolecule monitoring in recent years, as document 1 (Detection of biomolecular microarrays withoutfluorescent-labeling agents, SPIE 5328 (121), 2004) introduced, adopting diameter in experiment is the hot spot irradiation sample of 3 μ m, receive reflected light with a silicon photoelectric diode, light path is fixed in the experimentation, and sample is done two dimension and moved.Whole device is included in the input path system of LASER Light Source, light ball modulator, general gram box (Pockels cell) and the lens composition of the P polarization that order is provided with on the light path, the emitting light path part and the data acquisition and processing (DAP) system that are made up of lens, analyzer and photodetector adopt in the document that this device has carried out respectively under not having fluorescently-labeled condition that the biomolecule arrays synthetic reaction detects, the detection of microarray in the detection by quantitative of the optical property of microarray and the aqueous solution.The time that the monitoring that this kind monitoring device reacts for many array element biomolecule microarry needs is long, and movement of sample has been brought the system noise that is difficult to avoid to monitoring result.
Summary of the invention:
The objective of the invention is to overcome the defective of existing OIRD biomolecule monitoring technology, thus provide a kind of need not mobile example, the device of monitoring time is short, system noise is low high-throughout monitoring micro-array biomolecule reaction by light reflection difference method.
The device of high-throughout monitoring micro-array biomolecule reaction by light reflection difference method provided by the invention, comprise input path, sample stage, emitting light path and data acquisition and processing (DAP) system, described input path comprises laser instrument, the polarizer, light ball modulator and phase shifter, and wherein order is provided with the polarizer, light ball modulator and phase shifter on the light path of laser instrument output light the place ahead; Described emitting light path comprises analyzing analyser and photoelectric signal converter, outgoing beam the place ahead after the reflection of the sample on the sample stage is settled described analyzing analyser and photoelectric signal converter in proper order, described photoelectric signal converter is electrically connected to described data acquisition and processing (DAP) system, it is characterized in that
Also comprise the parallel beam expand device that is arranged between phase shifter and the sample stage in the described input path;
Also comprise a mechanical joint, this mechanical joint is by two risers that are vertically fixed on the sample stage, with a flat aluminium alloy plate of accepting incident light, with a flat aluminium alloy plate that is used to reflect emergent light, have the circular arc fixing threaded hole on described two flat aluminium alloy plates and two risers, by a bolt two oblique being fixed on the riser of flat aluminium alloy plate are formed, described screw is the quadrant arcuation, and aluminium alloy plate can be reconciled between 0-90 °;
Described input path is arranged on the flat aluminium alloy plate of accepting incident light and described emitting light path is arranged on the flat aluminium alloy plate that is used to reflect emergent light;
Described photoelectric signal converter comprises silicon photodiode array, on-off circuit, first lock-in amplifier and second lock-in amplifier, and wherein said silicon photodiode array output terminal is electrically connected to the input end of on-off circuit, first lock-in amplifier and second lock-in amplifier respectively.
In above-mentioned technical scheme, described data acquisition and processing (DAP) system comprises BNC adapter, data collecting card and data processing equipment; Wherein
Described BNC adapter receives described first lock-in amplifier and the second lock-in amplifier output signal, and the signal of phase-locked output is carried out the subchannel processing, and the signal that each lock-in amplifier is exported all is divided into amplitude information and two parts of phase information;
Described data collecting card is gathered the data of BNC adapter output, and sends data processing equipment to;
Described data processing equipment is stored, is analyzed and handle the data that the data capture card sends.
In above-mentioned technical scheme, described parallel beam expand device is the beam broadening lens combination, described beam broadening lens combination comprises collimation lens barrel and two continuous zoom expander lens that are separately positioned on described collimation lens barrel two ends, by regulating the length of collimation lens barrel, can adjust the diameter of irradiating light beam xsect.
In above-mentioned technical scheme, described input path is connected with the mechanical joint of emitting light path by the adjustment beam incident angle.
In above-mentioned technical scheme, described laser instrument is that wavelength is the linear polarization He-Ne laser instrument of 632.8nm.
In above-mentioned technical scheme, described light ball modulator is modulating frequency Ω=50kHz.
In above-mentioned technical scheme, described sample stage is the stainless steel platform with three-dimensional manual adjustments that has the sample fixed mechanism.
Advantage of the present invention:
(1) the present invention adopts silicon photodiode array as catoptrical receiving element, when incoming laser beam incides on the microarray sample with angle pencil of ray through after a series of optical systems, reflected light is received by silicon photodiode array behind the analyzing analyser, when incident angle one timing, need not two-dimentional mobile example platform, only adopt silicon photodiode array just the light of biological sample surface reflection all can be received, so not only reduce the noise that mobile example brings, and improved signal to noise ratio (S/N ratio) greatly;
(2) adopt angle pencil of ray incident and silicon photodiode array to receive reflected light, make device of the present invention can be simultaneously carry out unmarked, fast high-flux monitoring, improved monitoring efficient greatly the multiple reaction of a plurality of array elements of microarray;
(3) apparatus structure of the present invention is simple, and easy operating can high flux be realized the monitoring to biomolecular reaction fast, without any need for labelled reagent, and high efficiency;
(4) the used device of the present invention can both have been bought on market, realizes easily;
(5) in addition, input path of the present invention system adopts mechanical joint to be connected with the emitting light path system, makes that the incident angle of light beam can arbitrarily adjustment between 0-90 °.
Description of drawings
The device of Fig. 1 high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of the present invention is formed synoptic diagram.
Reference numeral:
The 1-laser instrument; The 2-polarizer; The 3-light ball modulator; The 4-phase shifter;
The 5-parallel beam expand device; The 6-sample stage; 7-analyzing analyser; The 8-on-off circuit;
The 9-silicon photoelectric diode; 10-first lock-in amplifier; 11-second lock-in amplifier;
12-data acquisition and processing (DAP) system.
Embodiment
The present invention will be further explained and explanation below in conjunction with specific embodiment.
Embodiment 1
With reference to figure 1, make the device of a high-throughout monitoring micro-array biomolecule reaction by light reflection difference method.
This device is made up of input path, sample stage 6, emitting light path and data acquisition and processing (DAP) system, and wherein input path is included on laser instrument 1 output light the place ahead light path the polarizer 2, light ball modulator 3, phase shifter 4 and parallel beam expand device 5 are set in proper order; Described emitting light path is by analyzing analyser 7 and photoelectric signal converter 9, the incident beam that sends from input path passes through described analyzing analyser 7 and photoelectric signal converter through the reflection of the sample on the sample stage 6 back order, and described photoelectric signal converter is electrically connected to described data acquisition and processing (DAP) system 12.Described photoelectric signal converter is made up of silicon photodiode array 9, on-off circuit, first lock-in amplifier 10 and second lock-in amplifier 11, and wherein silicon photodiode array 9 output terminals are electrically connected to the input end of on-off circuit, first lock-in amplifier 10 and second lock-in amplifier 11 respectively.
Also adopt mechanical joint to regulate incident and emergent light in the device of present embodiment, this mechanical joint is by two risers that are vertically fixed on the sample stage, with a flat aluminium alloy plate of accepting incident light, with a flat aluminium alloy plate that is used to reflect emergent light, have the circular arc fixing threaded hole on described two flat aluminium alloy plates and two risers, by a bolt two oblique being fixed on the riser of flat aluminium alloy plate are formed, described screw is the quadrant arcuation, aluminium alloy plate can be reconciled between 0-90 °, and such conciliation has realized the adjusting of incident and emergent light.Above-mentioned input path is arranged on the flat aluminium alloy plate of accepting incident light and above-mentioned emitting light path is arranged on the flat aluminium alloy plate that is used to reflect emergent light.
What adopt in the device of present embodiment is that power is 4mW, wavelength is that the He-Ne laser instrument of 632.8nm is as light source, its bright dipping aperture is 3mm, becomes the p polarized light that the polarization direction is parallel to the plane of incidence after laser instrument 1 emitting laser bundle is proofreaied and correct the polarization direction via the polarizer (New Focus5524) 2; Then pass through light ball modulator (Hinds Instrument PEM90 again
TM) 3 producing the periodic modulation between p and s polarization state, modulating frequency is 50KHz; Is the scalable phase compensation that phase shifter 4 that the quartzy multistage half-wave plate of 25.4mm constitutes is introduced between p and s polarization state from the polarization-modulated light of light ball modulator 3 outgoing via a diameter; Then by inciding on the sample stage 6 after with the laser beam broadening as the beam broadening lens combination of parallel beam expand device 5, described beam broadening lens combination is made up of collimation lens barrel and two continuous zoom expander lens that are separately positioned on described collimation lens barrel two ends, the adjustable length of described collimation lens barrel, its total length is 134mm to the maximum, and the applicable wavelengths scope 450-680nm of beam broadening lens combination, expansion beam ratio are that 5X-6X, launching spot and emergent light spot diameter are respectively 4mm and 24mm; Described sample stage 6 adopts the stainless steel table top of buying on the market, the platform that can realize three-dimensional manual adjustments, and degree of regulation is 0.01mm, and three-dimensional stroke is 50mm;
The reflected light of sample surfaces reflection is received by silicon photodiode array 9 after analyzing analyser 7 (CVI Laser CPAD-10.0-425-675) is regulated; First lock-in amplifier 10 and second lock-in amplifier 11 (Stanford Research Systems SR830DSP) are electrically connected to silicon photodiode array 9 respectively, and described silicon photo diode array 9 is 1.0mm by photosensitive area
2Square high-speed induction silicon photo diode (BPX-65) form 30 * 30 two-dimensional arraies.Because silicon photodiode array 9 has several silicon photoelectric diodes, for the light signal that each silicon photoelectric diode is received can both be analyzed with first lock-in amplifier 10 and second lock-in amplifier 11, on silicon photodiode array 9, be electrically connected an on-off circuit, on-off circuit in the present embodiment is a relay commutation circuit 8, and one-level harmonics and secondary harmonics that the light signal that each silicon photoelectric diode is accepted changes into behind the electric signal can be read by the data acquisition and processing (DAP) system successively;
Data acquisition and processing (DAP) system 12 in this device adopts computing machine, this machine is calculated and is disposed data collecting card (PCI-6220), the capture program that BNC adapter (BNC-2110) and LabVIEW write, the electric signal of first lock-in amplifier 10 and 11 outputs of second lock-in amplifier at first carries out subchannel via the BNC adapter to be handled, the signal of each lock-in amplifier output all is divided into amplitude information and two parts of phase information, then the amplitude information and the phase information of each lock-in amplifier are sent into the data collecting card collection respectively, at last the data that collect are sent to the data processor that adopts LabView to write and store, analyze and handle.
Adopt device of the present invention when monitoring accuracy is 2um, can within 2 minutes, finish once monitoring to the biological sample of 20 * 20 to 200 * 200 biochip arrays, wherein the diameter of array element is 100um, and spacing is 100um, and monitoring sensitivity reaches 2 * 10
-6Rad.
Embodiment 2
In the present embodiment, it is the general gram box of Cleveland Crystal IMPACT10 that phase shifter 4 adopts model, and other are identical with embodiment 1, and the device sensitivity of present embodiment is compared embodiment 1 can improve 1 order of magnitude.
Embodiment 3
Adopt on-off circuit 8 to replace the relay commutation circuit in the present embodiment, other is identical with embodiment 1, and the same used time of biochip sample of this device monitoring is compared embodiment 1 and can shorten 0.5 minute.
Claims (7)
1. the device of a high-throughout monitoring micro-array biomolecule reaction by light reflection difference method, comprise input path, sample stage, emitting light path and data acquisition and processing (DAP) system, described input path comprises laser instrument, the polarizer, light ball modulator and phase shifter, and wherein order is provided with the polarizer, light ball modulator and phase shifter on the light path of laser instrument output light the place ahead; Described emitting light path comprises analyzing analyser and photoelectric signal converter, outgoing beam the place ahead after the reflection of the sample on the sample stage is settled described analyzing analyser and photoelectric signal converter in proper order, described photoelectric signal converter is electrically connected to described data acquisition and processing (DAP) system, it is characterized in that
Also comprise the parallel beam expand device that is arranged between phase shifter and the sample stage in the described input path;
Also comprise a mechanical joint, this mechanical joint is by two risers that are vertically fixed on the sample stage, with a flat aluminium alloy plate of accepting incident light, with a flat aluminium alloy plate that is used to reflect emergent light, have the circular arc fixing threaded hole on described two flat aluminium alloy plates and two risers, by a bolt two oblique being fixed on the riser of flat aluminium alloy plate are formed, described screw is the quadrant arcuation, and aluminium alloy plate can be reconciled between 0-90 °;
Described input path is arranged on the flat aluminium alloy plate of accepting incident light and described emitting light path is arranged on the flat aluminium alloy plate that is used to reflect emergent light;
Described photoelectric signal converter comprises silicon photodiode array, on-off circuit, first lock-in amplifier and second lock-in amplifier, and wherein said silicon photodiode array output terminal is electrically connected to the input end of on-off circuit, first lock-in amplifier and second lock-in amplifier respectively.
2. by the device of the described high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of claim 1, it is characterized in that described data acquisition and processing (DAP) system comprises BNC adapter, data collecting card and data processing equipment; Wherein,
Described BNC adapter receives described first lock-in amplifier and the second lock-in amplifier output signal, and the signal of phase-locked output is carried out the subchannel processing, and the signal that each lock-in amplifier is exported all is divided into amplitude information and two parts of phase information;
Described data collecting card is gathered the data of BNC adapter output, and sends data processing equipment to;
Described data processing equipment is stored, is analyzed and handle the data that the data capture card sends.
3. press the device of the described high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of claim 1, it is characterized in that, described parallel beam expand device is the beam broadening lens combination, described beam broadening lens combination comprises collimation lens barrel and two continuous zoom expander lens that are separately positioned on described collimation lens barrel two ends, by regulating the length of collimation lens barrel, can adjust the diameter of irradiating light beam xsect.
4. by the device of the described high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of claim 1, it is characterized in that described sample stage is the stainless steel platform with three-dimensional manual adjustments that has the sample fixed mechanism.
5. by the device of the described high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of claim 1, it is characterized in that described laser instrument is that wavelength is the linear polarization He-Ne laser instrument of 632.8nm.
6. by the device of the described high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of claim 1, it is characterized in that described light ball modulator is modulating frequency Ω=50kHz.
7. by the device of the described high-throughout monitoring micro-array biomolecule reaction by light reflection difference method of claim 1, it is characterized in that described on-off circuit is relay commutation circuit or pulse-switching circuit.
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| CNA2006101135764A CN101153869A (en) | 2006-09-30 | 2006-09-30 | Device for high-throughout monitoring micro-array biomolecule reaction by light reflection difference method |
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| CNA2006101135764A CN101153869A (en) | 2006-09-30 | 2006-09-30 | Device for high-throughout monitoring micro-array biomolecule reaction by light reflection difference method |
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| CN102192879A (en) * | 2010-03-17 | 2011-09-21 | 中国科学院物理研究所 | Unmarked high throughput detecting device of biological chip by using light reflection differential method |
| CN102262052A (en) * | 2010-05-26 | 2011-11-30 | 中国科学院理化技术研究所 | Laser confocal obliquely-incident ellipsometric high-throughput biomolecular reaction imaging detection device |
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| CN102192879A (en) * | 2010-03-17 | 2011-09-21 | 中国科学院物理研究所 | Unmarked high throughput detecting device of biological chip by using light reflection differential method |
| CN102192879B (en) * | 2010-03-17 | 2013-05-22 | 中国科学院物理研究所 | Unmarked high throughput detecting device of biological chip by using light reflection differential method |
| CN102262052A (en) * | 2010-05-26 | 2011-11-30 | 中国科学院理化技术研究所 | Laser confocal obliquely-incident ellipsometric high-throughput biomolecular reaction imaging detection device |
| CN102262052B (en) * | 2010-05-26 | 2012-12-26 | 中国科学院理化技术研究所 | Laser confocal obliquely-incident ellipsometric high-throughput biomolecular reaction imaging detection device |
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| CN105675542B (en) * | 2015-08-24 | 2018-09-28 | 温州大学 | A kind of device and method of quick identification gutter oil |
| CN105675542A (en) * | 2015-08-24 | 2016-06-15 | 温州大学 | Device and method for rapid identification of swill-cooked dirty oil |
| CN105547947A (en) * | 2015-09-23 | 2016-05-04 | 中国石油大学(北京) | Light reflection difference device and method for detecting PM2.5 |
| CN105628620A (en) * | 2015-09-23 | 2016-06-01 | 中国石油大学(北京) | Light reflex difference device and method for water flooding dynamic detection |
| CN105548084A (en) * | 2015-09-23 | 2016-05-04 | 中国石油大学(北京) | Light reflection difference device and method for detecting rock stress sensitivity |
| CN105717018A (en) * | 2015-09-23 | 2016-06-29 | 中国石油大学(北京) | Device and method using light reflection difference to detect rock pore structure |
| CN105717049A (en) * | 2015-09-23 | 2016-06-29 | 中国石油大学(北京) | Light reflection difference device and method for detecting oil content of oil shale |
| CN105738290A (en) * | 2015-09-23 | 2016-07-06 | 长江大学 | Device and method for representing and recognizing crude oil from different producing places through light reflex difference method |
| CN105738321A (en) * | 2015-09-23 | 2016-07-06 | 长江大学 | Light reflection difference method for detecting illegal cooking oil |
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Application publication date: 20080402 |