KR101749623B1 - Multi-channel optical sensing apparatus using surface plasmon resonance induced fluorescence signal enhancement - Google Patents

Abstract

The present invention relates to a multi-channel optical sensing device using fluorescence detection signal amplification by surface plasmon resonance, and more particularly, to a multi-channel optical sensing device using amplification of a fluorescence signal by surface plasmon resonance of light, Fluorescence detection by surface plasmon resonance, which can detect not only the concentration but also the detection substance having a low concentration, and can detect fluorescence signals having different colors in multiple channels by distinguishing the fluorescent substances accommodated in the channels Channel optical sensing apparatus using signal amplification.
To this end, the present invention provides a multi-channel optical sensing device using a fluorescence detection signal amplification by a surface plasmon resonance phenomenon including a chamber, a light source part, and a metal nano thin film. The chamber accommodates a buffer solution in which a detection substance and a fluorescent substance to be bound to the detection substance are mixed, the light source section is disposed in an upper part of the chamber, and the light is irradiated, and the metal nano thin film is disposed so as to generate surface plasmon resonance .

Description

[0001] The present invention relates to a multi-channel optical sensing apparatus using surface plasmon resonance

The present invention relates to a multi-channel optical sensing device using fluorescence detection signal amplification by surface plasmon resonance, and more particularly, to a multi-channel optical sensing device using amplification of a fluorescence signal by surface plasmon resonance of light, Fluorescence detection by surface plasmon resonance, which can detect not only the concentration but also the detection substance having a low concentration, and can detect fluorescence signals having different colors in multiple channels by distinguishing the fluorescent substances accommodated in the channels Channel optical sensing apparatus using signal amplification.

Surface plasmons (SPs) are also called surface plasmon polaritons (SPPs) or plasmon surface polarites (PSPs).

Surface plasmons are generally the collective vibration of conduction band electrons propagating along the interface of a metal with a negative dielectric function (ε '<0) and a medium with a positive (ε'> 0) collective oscillation phenomenon, which is excited as a result of interaction with light (more specifically, electromagnetic waves) and has an enhancement size larger than incident light, and exponentially decaying as it moves away from the interface in the vertical direction evanescent wave).

In other words, surface plasmon resonance (SPR) phenomenon can be defined as a unique phenomenon caused and observed as a result of the interaction between photons and noble metal nanoscale.

Fluorescence signals, which are widely used in almost all fields of medicine, engineering, environment, energy, etc., are electromagnetic waves (light) emitted when fluorescent molecules are excited by the light in the outside of the ultraviolet ray or visible ray region, The emission light is an electromagnetic wave with lower photon energy (longer wavelength) than the excitation light source. The electromagnetic wave generated is emitted in the direction of 4π solid angle, ie, in all radial directions, and the efficiency of the emitted electromagnetic wave which can be detected by the condensing is generally less than 1%.

When using very expensive condensing optics, it can go up to 10%, but it is very expensive and complicated to apply to general fluorescence experiments and fluorescence. Therefore, when detecting fluorescence with various photodetectors such as a charge coupled device (CCD), a complementary metal oxide semiconductor (CMOS) photodetector, and a semiconductor photodiode, the amount of condensed fluorescence is determined by the dark current, And the signal-to-noise ratio is generally lowered.

To overcome this, we have to use a very sensitive and expensive photodetector, such as a photo-muliplier tube (PMT), to detect fluorescence in a dark, enclosed area such as a dark room, or to detect fluorescence in a non- I will not.

In addition, it has been proposed that the fluorescence detection efficiency is improved by the surface plasmon resonance (SPR) and the generated fluorescence signal itself can be amplified, and some related technologies have been experimentally realized. However, There is a problem that it is difficult to use various fluorescence detection techniques and related optical technologies such that the fluorescence detection efficiency by the SPR and the fluorescence amplification technique can be used in many fluorescence applications such as energy.

Korean Patent Laid-Open Publication No. 10-2016-0040478 (Title: Surface plasmon resonance sensor system capable of real-time analysis of a sample to be analyzed, and analysis method using the same) Published Apr. 14, 2016

A problem to be solved by the present invention is to detect the concentration of a detection substance contained in a buffer solution as well as to detect a detection substance having a low concentration by amplifying a fluorescent signal by surface plasmon resonance of light, Channel optical sensing apparatus using fluorescence detection signal amplification by surface plasmon resonance phenomenon capable of detecting fluorescence signals having different colors in multiple channels by distinguishing fluorescent substances accommodated therein.

According to an aspect of the present invention, there is provided a light sensing apparatus using surface plasmon resonance, comprising: a chamber in which a buffer solution containing a detection substance and a fluorescent substance bound to the detection substance is accommodated; A light source unit disposed above the chamber to emit light; And a metal nano thin film disposed on the bottom of the chamber so as to generate surface plasmon resonance, wherein the fluorescent material includes a first fluorescent material and a second fluorescent material having different emission wavelengths from the light emitted from the light source, The lower portion of the chamber is divided into a first channel in which the first fluorescent material is accommodated and a second channel in which the second fluorescent material is accommodated, the metal nano thin film includes a first metal thin film layer disposed under the first channel, And a second metal thin film layer disposed under the second channel, wherein the first metal thin film layer and the second metal thin film layer have different thicknesses depending on a divergent wavelength of the first fluorescent material and the second fluorescent material, The first fluorescent material and the second fluorescent material emit light by the light emitted from the light source portion to generate a fluorescent signal, And a fluorescent signal generated by light emitted from the second fluorescent material is amplified by surface plasmon resonance generated by the buffer solution, the first metal thin film, and the second metal thin film. A multi-channel optical sensing device using a fluorescence detection signal amplification by a fluorescence detection signal is provided.

The metal thin film layer may be formed thick as the divergent wavelength of the fluorescent material becomes longer.

The material constituting the metal thin film layer may be gold or silver.

Wherein the light source unit includes a first light source having a first excitation wavelength for exciting the first fluorescent material and for emitting a first excitation light having the first excitation wavelength and a second excitation light for exciting the second fluorescent material, And a second light source for irradiating the second excitation light having the second excitation wavelength.

The multi-channel optical sensing apparatus using the fluorescence detection signal amplification by the surface plasmon resonance phenomenon according to the present invention has the following effects.

First, the fluorescence signal generated by the light source is amplified through surface plasmon resonance, thereby enabling detection of a low-concentration detection substance.

Second, various fluorescent materials are accommodated in the respective channels and irradiated with light, whereby the fluorescence signal can be detected in multiple channels.

Third, by optimizing the thickness of the metal nano-thin film for generating the surface plasmon resonance according to the respective fluorescent materials accommodated in the channel to optimize the plasmon surface wave thickness, it is possible to expand the amplification range of the fluorescent signal of the fluorescent material There is an advantage.

1 is a schematic view of a multi-channel optical sensing device using fluorescence detection signal amplification by surface plasmon resonance according to a first embodiment of the present invention.
2 is a view for explaining the thickness of a metal thin film according to a fluorescent material.
3 is a schematic view of a multi-channel optical sensing device using fluorescence detection signal amplification by surface plasmon resonance according to a second embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention in which the above-mentioned problems to be solved can be specifically realized will be described with reference to the accompanying drawings. In describing the embodiments, the same names and the same symbols are used for the same configurations, and additional description thereof will be omitted in the following.

A first embodiment of a multi-channel optical sensing device using fluorescence detection signal amplification by surface plasmon resonance phenomenon according to the present invention will be described with reference to FIGS. 1 and 2. FIG.

As shown in FIG. 1, the multi-channel optical sensing device using the fluorescence detection signal amplification by the surface plasmon resonance phenomenon of the present invention includes a chamber 120, a light source 110, and a metal nano-thin film.

The chamber 120 receives the buffer solution L in which the detection material and the fluorescent material to be coupled to the detection material are mixed and the buffer solution L is applied to the dielectric material so that surface plasmon resonance can occur at the interface with the metal nanofiltration film. It plays a role.

Specifically, the lower region of the chamber 120 includes a first channel 120a, a second channel 120b, and a second channel 120b in which fluorescent materials having different detection materials and different emission wavelengths, And is divided into three channels 120c.

However, the number of channels may be changed according to the type of fluorescent material to be detected.

Specifically, the first fluorescent material 141 is received in the first channel 120a and coupled to the first detection material D1. In the second channel 120b, the emission wavelength is longer than that of the first fluorescent material 141 The second fluorescent material 142 is received and combined with the second detection material D2 and the third fluorescent material 143 having a longer wavelength of emission than that of the second fluorescent material 142 is accommodated in the third channel 120c And is combined with the third detecting material (D3).

The first channel 120a, the second channel 120b and the third channel 120c may be formed of the first fluorescent material 141, the second fluorescent material 141, 142 and the third fluorescent material 143 emit fluorescence signals of different colors at the same time.

In this specification, the first fluorescent material 141 is FAM (5-carboxyfluorescein), the second fluorescent material 142 is SYBR green I, the third fluorescent material 143 is Fluorescein isothiocyanate (FITC) The divergent wavelength of the material 141 is 518 nm, the divergent wavelength of the second fluorescent material 142 is 522 nm, and the divergent wavelength of the third fluorescent material 143 is 525 nm.

Of course, as the fluorescent material accommodated in the channels partitioned according to the type and number of the detection substances to be detected, not only the above-described fluorescent material but also fluorescent materials having different emission wavelengths such as Cy3, Cy5, and EtBr may be used.

The detection material may be composed of other kinds of biomaterials such as DNA or RNA. The fluorescence material generates a fluorescence signal by the light irradiated by the light source unit 110 so that the type and concentration of the detection substance can be grasped do.

The light source unit 110 is disposed on the chamber 120, and the fluorescent material generates a fluorescent signal by irradiating the fluorescent material with light. The light source unit 110 is composed of white light including all wavelengths of light, Even if you have it, you can make the fluorescent material generate the fluorescent signal.

Therefore, the fluorescence signal can be detected without using a light source having different wavelengths according to the divergent wavelength of the fluorescent material.

The metal nano thin film is disposed under the chamber 120, and surface plasmon resonance occurs between the buffer solution L serving as a dielectric and the metal nano thin film, and the metal nano thin film is placed under the first channel 120a A second metal thin film layer 132 disposed under the second channel 120b and a third metal thin film layer 133 disposed under the third channel 120c.

The first metal thin film layer 131, the second metal thin film layer 132, and the third metal thin film layer 133 have different thicknesses. The first metal thin film layer 131, the second metal thin film layer 132, and the third metal thin film layer 133 are made of gold (Au) The thickness of the metal nano-thin film varies depending on the refractive index of the buffer solution L and the divergent wavelength of each fluorescent material accommodated in the first channel 120a to the third channel 120c.

The adjustment of the thickness of the metal nanofiltration layer according to the type of the fluorescent material accommodated in the first channel 120a through the third channel 120c is performed by the surface plasmon resonance occurring between the buffer solution L and the metal nanofiltration layer And to increase the Q-factor (quality factor) that maximizes the plasmon surface wave thickness (t).

Specifically, when the surface plasmon Q-factor is enhanced, the propagation length of the plasmon surface wave and the intensity of the propagation signal are increased at the same time, thereby increasing the amplification range of the fluorescent signal of the fluorescent material. Thereby increasing the amplification of the signal.

As shown in FIG. 2, when the metal nano thin film is made of gold and the first fluorescent material 141 to the third fluorescent material 143 are made of a fluorescent material having the above-mentioned divergent wavelength, The longer the divergence wavelength, the thicker the metal nano-thin film and the more optimized the Q-factor.

That is, when the divergent wavelength of the second fluorescent material 142 is longer than the divergent wavelength of the first fluorescent material 141 and the divergent wavelength of the third fluorescent material 143 is longer than the divergent wavelength of the second fluorescent material 142 And a second channel 120b disposed below the second channel 120b in which the second fluorescent material 142 is received than a thickness of the first metal film layer 131 disposed below the first channel 120a in which the first fluorescent material 141 is accommodated. The third metal thin film layer 133 is formed to be thicker than the second metal thin film layer 132 and disposed below the third channel 120c in which the third fluorescent material 143 is contained than the thickness of the second metal thin film layer 132. [ And the thickness is formed thick.

Accordingly, the first channel 120a to the third channel 120c are optimized for the plasmon surface wave thickness t by the optimized Q-factor, and the fluorescence of the first fluorescent material 141 to the third fluorescent material 143 As the amplification range of the signal is increased, the detection efficiency is improved as compared with the general fluorescence signal detection, so that accurate sensing is achieved.

In addition, as the fluorescence signal of the fluorescent material is amplified, the sensing material can be sensed even when the detection substance is contained in the buffer solution L at a low concentration, and even when the concentration difference is small in each channel, The difference can be easily grasped so that more accurate sensing can be achieved.

Meanwhile, the first detection material D1, the second detection material D2, and the third detection material D3 are received in the first channel 120a, the second channel 120b and the third channel 120c, respectively The same fluorescent material may be bonded to the first detecting material D1, the second detecting material D2 and the third detecting material D3, and the first detecting material D1 to the third detecting material D3, A fluorescent material having a different divergent wavelength may be combined.

When different detection materials are accommodated in the first to third channels 120a to 120c and the same fluorescent material is coupled to each of the detection materials, It is possible to distinguish the fluorescence signal as a function of the position of the fluorescence signal using a lens or the like.

In addition, when different detection materials are accommodated in the first to third channels 120a to 120c and fluorescent materials having different emission wavelengths are coupled to the respective detection materials, The directionality of the fluorescence signal is different from each other. Accordingly, it is possible to separately detect the fluorescence signal for each channel using a lens or the like.

In particular, when a plurality of detection materials are accommodated in one channel and a fluorescent material having different divergent wavelengths is coupled to each of the detection materials, the directionality of the divergent fluorescent signal changes depending on the divergent wavelength, A plurality of detection materials can be separately detected in one channel.

As shown in FIG. 3, a second embodiment of a multi-channel optical sensing device using fluorescence detection signal amplification by surface plasmon resonance according to the present invention will now be described.

The chamber according to the present embodiment is divided into two or more channels, and fluorescent materials having different emission wavelengths are accommodated in the respective channels and are combined with different detection materials. Since the thickness of the metal nano-thin film disposed on the substrate 100 is substantially the same as that of the first embodiment, detailed description thereof will be omitted.

However, the first to third channels 220a to 220c according to the present embodiment are different from the first to third channels 120a to 120c of the first embodiment only in the lower region of the chamber 220 It is not partly partitioned, but has an entirely divided and independent space.

Also, the light source unit according to the present embodiment includes a first light source unit 211, a second light source unit 212, and a third light source unit 213.

The first light source unit 211 has a first excitation wavelength for exciting the first fluorescent material 241 and is disposed on the first channel 120a to excite the first fluorescent material 241 to the first excitation wavelength So that the first fluorescent material 241 generates a fluorescent signal.

The second light source part 212 has a second excitation wavelength for exciting the second fluorescent material 242 and is disposed on the second channel 120b so as to be incident on the second fluorescent material 242 at a second excitation wavelength, So that the second fluorescent material 242 generates a fluorescent signal.

The third light source unit 213 has a third excitation wavelength for exciting the third fluorescent material 243 and is disposed on the third channel 120c to excite the third fluorescent material 243 to the third excitation wavelength, So that the third fluorescent material 243 generates a fluorescent signal.

As described above, by arranging the light source portions having different excitation wavelengths according to the divergent wavelengths of the first to third fluorescent materials 241 to 243 in the respective channels, the fluorescent signals of different colors are simultaneously displayed Thus, the concentration of the detection substance can be grasped simultaneously in multiple channels.

As described above, the present invention is not limited to the above-described specific preferred embodiments, and various changes and modifications may be made by those skilled in the art without departing from the scope of the present invention as claimed in the claims. And such variations are within the scope of the present invention.

110: light source part 211: first light source
212: second light source 213: third light source
120, 220: chambers 120a, 220a: first channel
120b, 220b: second channel 120c, 220c: third channel
131, 231: first metal thin film layer 132, 232: second metal thin film layer
133, 233: third metal thin film layer 141, 241: first fluorescent material
142, 242: Second fluorescent material 143, 243: Third fluorescent material
D1: first detection substance D2: second detection substance
D3: third detection substance L: buffer solution

Claims (4)

A chamber in which a buffer solution containing a detection substance and a fluorescent substance bound to the detection substance is accommodated;
A light source unit disposed above the chamber to emit light;
And a metal nano thin film disposed to generate surface plasmon resonance in the lower portion of the chamber,
Wherein the fluorescent material includes a first fluorescent material and a second fluorescent material having different divergent wavelengths from the light emitted from the light source,
The lower portion of the chamber is divided into a first channel in which the first fluorescent material is accommodated and a second channel in which the second fluorescent material is accommodated,
Wherein the metal nano thin film includes a first metal thin film layer disposed under the first channel and a second metal thin film layer disposed under the second channel,
Wherein the first metal thin film layer and the second metal thin film layer have different thicknesses depending on a divergent wavelength of the first fluorescent material and the second fluorescent material,
The first fluorescent material and the second fluorescent material are excited by the light emitted from the light source unit to generate a fluorescent signal, and at the same time, a fluorescent signal due to the light emitted from the first fluorescent material and the second fluorescent material, And the second metal thin film is amplified by surface plasmon resonance generated at the interface between the first metal thin film and the second metal thin film by using the fluorescence detection signal amplification by the surface plasmon resonance phenomenon. The method according to claim 1,
Wherein the metal thin film layer is formed thick as the divergent wavelength of the fluorescent material becomes longer. The multi-channel optical sensing device using the fluorescence detection signal amplification by the surface plasmon resonance phenomenon. 3. The method of claim 2,
Wherein the metal thin film layer is made of gold or silver. The multi-channel light sensing device using fluorescence detection signal amplification by surface plasmon resonance phenomenon. The method according to claim 1,
Wherein the light source unit includes a first light source having a first excitation wavelength for exciting the first fluorescent material and for emitting a first excitation light having the first excitation wavelength and a second excitation light for exciting the second fluorescent material, And a second light source for irradiating the second excitation light having the second excitation wavelength. The multi-channel optical sensing apparatus using the fluorescence detection signal amplification by the surface plasmon resonance phenomenon.

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