TW393575B - Optical heterodyne, phase sensitive surface plasma wave biosensor - Google Patents

Abstract

In conventional method, the surface plasma resonance (SPR) biosensor detects the intensity change of the reflected P polarized wave (TM wave) when the surface plasma wave is being generated. Simultaneously, there is a phase change of the reflected P polarized wave as well. Besides, the phase change of the reflected P polarized wave is significantly enhanced in comparison with the amplitude change when the incident angle of the laser beam is around the SPR angle. Once the antibody-antigen combination is formed and immobilized on the metal thin film of SPR generator, the extra phase change of the reflected P polarized light is then observed. A common path Mach-zehnder optical heterodyne interferometer, which provides higher stability and sensitivity of the phase measurement, is therefore suggested to sense the concentration of the antigen of the test biological substance. A novel SPR phase sensitive biosensor with the feature of optical common path configuration and optical heterodyne technique is proposed in this patent.

Description

V. Description of the Invention (/ A7 B7 Traditionally, it takes at least three days to detect the pathogenic Zhao (bacteria or virus) by ELISA method, and it must be completed by washing-reaction-washing steps. If fiber optic biosensors are used, it can greatly The test time is shortened to 20 minutes, however, iabei fluorescent agent is required to perform it, and there is a limitation on the sensitivity of the test. The latest method is to use SPR (surface plasma resonance) to have a fast test at the same time. Advantages of using no fluorescent agent '° The basic principle of SPR is mainly under the condition of measuring the total reflection of p wave (p polarized wave or TM wave) in the diamond mirror; Kgsin0 = Ksp; Kg = 2Vi ^ Ks C ω is the frequency of incident laser light εο, ει, (please read “Notes on the back of the page before filling in this page) —HI —--- 1r Printed by U. * \ Gl + £ 2j is the dielectric coefficient of different media. The P-wave reflection intensity changes due to the excited surface plasma wave (SPW) (as shown in the first figure), and the angle of incident light that generates SPR resonance is determined. ΘΓ, • this way The test has been widely used in the biomedical or chemical substance determination test.) · Ranxi This method uses the change in reflected light intensity to measure the value ', which is low in sensitivity. There are also chemical substance analyses using SPR methods combined with electrochemical electro-sensors to measure current. The present invention provides a new plasma common path architecture and heterodyne interference phase detection technology for surface plasma wave biosensors (that is, biosensors) to measure the phase change of the surface plasma wave p reflected wave. To replace the change in the intensity of the reflection first, and then obtain the concentration of the test object in the solution in real time. However, because the phase detection is very susceptible to interference from changes in the external environment, the optical common path architecture is useful for this paper. Two kneading rates in China (cps > A as I® · (210X297 mm) ----- install, '------ order .--------- 竽 ----- --1 ----------- A7 ------ B7___ V. Description of the invention (厶) ~---Π '| The external environment is immune to changes A 丨 The present invention combines surface electricity n Heterodyne interference technology and optical j common road structure 'Using her technology to replace _degrees, the system recording degree can be large I | amplitude increase β Ρ ί hl

The optical architecture of the common path heterodyne interference phase detection surface plasma wave biosensor is as shown in the second figure. Polarized laser light (combined with P and s waves) (1) via chirped wave plate (2) and beam splitting | J

2 I I slice B & (3) is divided into two beams; the signal beam and the reference beam, the signal beam is complicated by the Wi-frequency driver (41)

° The Acousto-Optic modulator A0M (4) of this T causes the laser light to generate a frequency Wi offset, and the reference beam passes the A0M (4) and the acousto-optic driver (42) to generate a Wz frequency offset. And adjust the signal I light (Pr ^) to the surface plasma wave biosensor (SPR) component ⑹ The angle of incidence 0 (as shown in Figure ^) is greater than the critical angle to generate total reflection and is related to the reference beam (p2 + S2) The polarized light beam splitter pBS⑺ 丨 · coincides, in which the P! Wave and P2 wave cause heterodyne interference, and its beat frequency, 丨 At the same time, A wave and S2 wave also generate heterodyne interference, and the beat frequency is also M, Because Naibo eliminates the printing of cooperatives in the SPR component (6) of the Ministry of Economic Affairs' Intellectual Property Bureau employees

The phase change occurs when the surface plasma wave is excited. However, the wave does not change at all because it does not excite the surface plasma wave. The light detectors Dp (8) and Ds (8) pass through the filter (9), respectively. Measured heterodyne interference | P-wave (P! + P2) and S-wave (Si + SJ signals can be written as

Ip (^ Wt) = IPi + Pj = 2λΑρΛ ^ cos (AWt + Φρ, -φρ2) .............. (1)

V

Is (AWt) = ISi + S2 = cos (/ VWt + < |) S |-< |) S2) ......... . (2) This paper is applicable to China National Standard (cys) person 4 threats (210X297 mm) A7 B7. (4) V. Description of the invention (3) and input to the lock in amplifier (10) , Can directly read out the phase difference δ of ip (Awt) and Is (ZXWt) and the ratio of the reflected light intensity of heterodyne interference P wave and S wave. Where δ = [(φΡι -Φρ2)-(Φ51 -Φ52)] .............. ............. (3) Meaning V ^ P | ^ P2 Asi ^ S2 Since (ΦΡ2, Φ5ι 木 2) and (ΪΡ2, IS1, IS2) are constants, given by Eq ( 3) It can be known that the present invention can simultaneously obtain changes in the phase of the dagger wave φP1 and the amplitude Ip. In terms of heterodyne interference P-wave (ρι + ρ2) & S-wave (S! + S2), the second optical architecture forms a common path. In phase measurement, it is immune to the external environment. This greatly improves the phase test accuracy. In the third figure, the surface plasma plasma wave SPR optical module has a glass based plate (53) and a metal film (54) with a thickness of 500A on the surface such as gold (Au) or silver (Ag). The optical refractive index matching solution (index) The purpose of matching oil) (52) is to combine the diamond lens (51) with the glass-filled substrate (53) so that their refractive indices are the same. The surface of the metal film (54) is plated with an antibody (55) for testing bacteria or viruses and is The antigen (56) of the test bacterium or virus binds and has an effect, and forms an antigen-antibody conjugate (57) in the reactor (58). The generation of the surface plasma wave and the surface of the metal thin film due to the attachment of the antibody antibody body (57) changed the effective optical refractive index, which caused the phase and intensity of the reflected p-wave to change. At the same time, the changes in the phase and amplitude of the reflected heterodyne interference P wave are used for the paper size. The two Chinese standards (CNS) Α4 size (210X297 mm) (please read the precautions on the back before filling this page) . 粲. Central Laboratories, Ministry of Economic Affairs, Shellfish Consumer Cooperatives, Printed Oxygen Ministry of Economics, 橾 Institute for Real Labor, Consumers Cooperation, Du Yin 辇 A7 B7 V. Description of Invention (f) '-The rapid inspection of bacteria is the invention patent The main content of this invention patent has the following advantages: 1. Sensitive material; heterodyne interference technology is applied to her in real time, the amount is the most sensitive method of gamma in the field of modern optics ~ her innovative heterodyne Interferometric phase detection technology, the detection sensitivity is greatly improved than the method of measuring the intensity of reflected waves. 2. The present invention proposes a new type of optical architecture, the common secret of silk optics, which can greatly improve the degree of joy and recording of the phase side measurement, and is not affected by external interference. 3. The invention does not require fluorescer, which makes the inspection process fast and simple. 4. The present invention uses a phase-locked amplifier for instant measurement of phase and vibration. 5 · "Benfa Ake 1: Measure the size of the ρ wave of the reflection treacherous wave, reduce the size of the ρ wave, reflect the optical structure of the common path of the yarn, and simultaneously measure the amplitude ratio of the heterodyne interference P wave and S wave; ^, not subject to laser light intensity The unstable Ying Xin's brief description of the 囷 style and the illustration of the original drawing symbols are as follows. Winter people and encountered TSBI home (CNS) A4 size (210X297 mm) I !: Install II (Please read the precautions on the back before writing this page) * π A9 B9 C9 D9 Schematic ϋ) First Figure Schematic illustration of the plasma wave generation on the surface of the Rhombus mirror Figure 2 The common optical path heterodyne interference phase detection surface plasma wave biosensor architecture Figure 3 The surface plasma plasma wave biosensor SPR optical component diagram (please read the back first (Notes for re-margining system) Order-line 丨 丨. Printed on paper scales by the Intellectual Property Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperatives, using China National Standard #CNS (CNS) A4 (210X297 mm) Intellectual Property of the Ministry of Economic Affairs Bureau's consumer cooperation Du printed A9 B9 C9 D9 schematic cy symbol element description: (1) linearly polarized laser light source (2) λ / 2 wave plate (3) beam splitter (4) acousto-optic modulator ( acousto-optic modulator) (41) driver (driver 1) (42) driver (driver 2) (5) reflector (6) SPR component ⑺ polarization beam splitter (8) detector (9) filter (10) ) Lock-in amplifier (11) Personal computer (51) Rhombus (52) Refractive index matching solution (53) Glass substrate (54) Metal film (55) Test for bacterial body antibodies (56) Test bacterial substance antigen (57) antigen-antibody conjugate (5 8) reaction chamber (69? Glass. Split scale., (60 / metal dielectric _ (Please read the precautions on the back before Line drawing) -β i! _ This paper size is in Chinese National Standard (CNS) A4 (210X297 mm)

Claims (1)

393575 AS B8 C8 D8 VI. Patent Application Fan Yuan ⑴ ~~: 1. An optical common path heterodyne interference phase detection surface plasma wave biosensor, which can qualitatively and quantitatively determine the species or concentration of bacteria or viruses, The structure can be composed of a surface plasma wave optical component (6), a linearly polarized laser light source (1), an input / 2-wave plate (2), a two beam splitter (3), and two acousto-optic modulators (4). , Two drivers (41) (42),-polarization beam splitter (7), a reflecting mirror (5), two light detectors (8), two filters (9), and a phase-locked amplifier (1 〇) and a human electroluminescence (η) and other components, in the surface plasma wave optical components of the test bacteria or virus antigens and antibodies attached to the surface of the metal film of the test film after binding, the incident laser light P wave (TM (Wave) produces a surface plasma wave on a metal surface, which causes a change in the phase and amplitude of the reflected P wave; the heterodyne interference P wave (PJP2) and the heterodyne interference S wave (Si + Sg) form an optical common path, which is Immunity to environmental changes> The system has high phase and amplitude measurement stability, sensitivity and signal-to-noise ratio (SNR) j Phase-amplifiers can be used to simultaneously measure the phase and amplitude changes of the reflected heterodyne interference P waves to determine the concentration of the bacteria or virus to be tested; the phase-locked amplifier used can detect the phase by a phase detector Changing j-line polarized laser can use solid (including semiconductor) or gas-body laser, acousto-optic modulator can use phase modulator, electro-optical modulator (Electro-optical modulator) Laser light frequency offset j acousto-optic modulator can use mechanical vibration or rotating grating (grating) to generate incident laser light frequency offset 'laser light can be linearly polarized and mutually perpendicular laser light with different laser light frequency | laser light You can use / current modulation to generate semi-specific lasers with dual frequencies and linear polarizations. The SPR paper size uses the Chinese national kneading rate (CNS > A4 size (210X297 mm) ---- ----- ^! (Please read the notes on the back before filling in this page) Ordering Line Consumer Intellectual Property Co., Ltd. of the Intellectual Property Bureau of the Ministry of Economic Affairs & a 393575 VI. Patent application scope Hemispherical identical glass Material replacement, and the use of focused incident p-wave laser light instead of parallel laser beams to generate surface plasma waves. A single detector in the system can use a 2-D photo detector array to achieve Requirement to measure multiple different incident angles at the same time. 2. The optical common path heterodyne interference phase detection surface plasma wave biosensor described in the first item of patent application scope can absorb the chemical substances of the test object. The invention can be applied to the detection of chemical substances in gas or liquid. 11 — ^ — 1—111 111 线 —ί (please read and read the note on the back of the page before filling out this page) Intellectual Property Bureau, Ministry of Economic Affairs, Consumer Consumption Du printed 210X297 mm) Private paper wave scale 逋 用 中国 国 # Rate (CJ4S) 8 specifications (