KR102102773B1 - Fluorescence analyzing apparatus based portable micro-optical component - Google Patents

Abstract

A fluorescence detection device for on-site diagnosis using an ultra-small optical element includes a light source unit that irradiates light, a variable focus lens that focuses light emitted from the light source unit, and a MEMS scanner that converts light focused from the variable focus lens into surface light. , A sample mixed with the reagent is amplified, and a light detection unit that detects fluorescence reflected from the biochip as the biochip that receives the surface light incident through the MEMS scanner and the target material of the sample reacts with the reagent You can.

Description

Fluorescence detection device for field diagnosis using ultra-small optical elements {FLUORESCENCE ANALYZING APPARATUS BASED PORTABLE MICRO-OPTICAL COMPONENT}

The present invention relates to a fluorescence detection device for on-site diagnosis, and more specifically, a microfluidic biochip using a small amount of a dielectric sample with high intensity of light excited over the entire surface of a sample by uniformly irradiating a focused light source with a MEMS scanner. The present invention relates to a fluorescence detection device for on-site diagnosis using an ultra-small optical element with improved detection efficiency.

Molecular diagnostics include the detection of hereditary diseases in the medical and biological fields, the identification of genetic fingerprints, the diagnosis of infectious diseases, the cloning of genes, and the testing of paternity, And DNA computing, which are widely used for analysis and diagnostic purposes.

Among them, a polymerase chain reaction (PCR) device is developing an efficient method for real-time monitoring of PCR progress as well as efforts to improve PCR yield. The technology capable of grasping the PCR progress in real time is referred to as "real-time PCR". In the real-time PCR device, a temperature control device and a spectral fluorescence photometer are integrated, and a fluorescent material is introduced into a PCR chamber. It uses the technology to measure the optical signal generated by combining with the amplification product.

However, this requires a structure such as a separate light source module for activating the optical signal from the fluorescent material, a photodetector module for detecting the optical signal obtained from the amplified nucleic acid, and other reflectors for adjusting the optical path.

Republic of Korea Patent Publication No. 2013-0113072 relates to an absorbance measurement device, and relates to an absorbance measurement device capable of detecting an absorbed state of a sample injected into a well using a CMOS image sensor. However, this has a disadvantage that the heat transfer efficiency is low in the sample using the well plate, so it takes a long time to perform the polymerase chain reaction.

Republic of Korea Registered Patent No. 1306930 relates to a spectrophotometer for simultaneous measurement of fluorescence absorption, and includes a light transmission unit and a multi-channel detection unit to control the path of light irradiated from one excitation light source, and simultaneously transmits fluorescence and absorption optical signals. Disclosed is a photodetector for measuring. However, this shows a structure that requires a separate light transport unit having a complex appearance and a filter, an additional lens, etc. in order to increase the measurement sensitivity of light and control the progress path.

Therefore, the device for measuring real-time fluorescence and absorbance includes a structure that increases the heat transfer efficiency to shorten the reaction time of the sample, and it is a compact optical that can control the intensity and scanning area of the light scanned over the surface area of the sample without a separate light transfer unit. There is a need for a detection device having an element.

Republic of Korea Patent Publication No. 10-2010-0029868 Republic of Korea Patent Publication No. 2013-0113072 Republic of Korea Registered Patent No. 1306930

Accordingly, the technical problem of the present invention was conceived in this regard, and the object of the present invention is to configure a small-sized optical system, a variable focus lens and a MEMS scanner, and a microfluidic-based biochip using a small sample, which is suitable for on-site diagnosis. The present invention relates to a fluorescence detection device for on-site diagnosis to which a small optical element capable of providing a detection device is applied.

A fluorescence detection device for on-site diagnosis using an ultra-small optical element according to an embodiment for realizing the object of the present invention, a light source unit irradiating light, a variable focus lens converging light emitted from the light source unit, and the variable focus lens In the Memes scanner converting the focused light into surface light, the sample mixed with the reagent is amplified, and the biochip and the target material of the sample react with the reagent as the surface light incident through the MEMS scanner is amplified. , A photodetector for detecting fluorescence reflected from the biochip.

In one embodiment, the light source unit may include a plurality of lasers irradiating laser light of different wavelengths and a first transfer stage for selectively aligning the lasers to the variable focus lens.

In one embodiment, the variable focus lens is located in a direction in which the focused light is traveling, and may further include a focus control mirror that reflects the focused light and transmits it to the MEMS scanner.

In one embodiment, a scanning lens provided by the biochip by converting and diffusing the surface light incident from the MEMS scanner into parallel light may be further included.

In one embodiment, the temperature of the biochip may further include a temperature control unit to amplify the sample in the biochip.

In one embodiment, the temperature control unit may include a plurality of temperature control units for heating the biochips to different temperatures or different temperature ranges, respectively.

In one embodiment, the temperature control unit may further include a second transfer stage to selectively move the temperature control unit to the lower portion of the biochip.

In one embodiment, the temperature control unit may include an indium tin oxide (ITO) of a Paltier element or a transparent material that generates heat when a current is applied.

In one embodiment, the MEMS scanner may convert the incident light into surface light that reflects the entirety of the scanning lens while changing the angle of reflection of the incident light through rotation.

In one embodiment, the biochip is an inlet part for injecting a plurality of reagents mixed with a sample, a mixing channel in which the sample and reagents injected through the inlet part are mixed, and connected to the mixing channel to the sample And a mixing chamber for further mixing reagents, a moving channel in which the samples and reagents mixed in the mixing chamber move, a plurality of reaction chambers in which the mixed samples and reagents branched from the moving channel react, and the reaction chamber It may include an outlet (Outlet) for discharging the sample and reagent reacted in the field.

In one embodiment, the reaction chambers form a plurality of spaces diverged from the moving channel, and the spaces may form an area corresponding to the surface light incident from the MEMS scanner.

In one embodiment, the reagents may be placed in the reaction chambers to perform a fluorescence reaction with the amplified target material of the sample.

According to the present embodiments, by applying a variable focus lens and a MEMS scanner with low power consumption, and configuring a microfluidic biochip using a small amount of reagent, it is possible to provide a compact detection device suitable for on-site diagnosis.

In addition, by using a variable focus lens, it is possible to adjust the focal length of light incident on the sample surface, and it is possible to provide focused light having a high light intensity on the sample surface.

In addition, since the focused light source is evenly irradiated by the MEMS scanner, there is an advantage that the intensity of light excited over the entire surface of the sample is increased, and accordingly, it is possible to provide focused light without a mechanism that functions as an optical waveguide.

In addition, it is possible to maximize the heat transfer efficiency through a microfluidic biochip using a small amount of reagent, thereby maximizing the polymerization element chain reaction (PCR) and reducing the time.

In this case, in order to perform detection of the target material in a larger area by a light source provided as a surface light source, a plurality of spaces may be formed in the reaction chambers, thereby relatively quickly and easily reacting with various samples. Can be detected.

In addition, considering the difference in amplification temperature according to a sample, by providing a plurality of temperature control units for heating at different temperatures or different temperature ranges, it is possible to provide optimal amplification conditions.

1 is a configuration diagram showing a fluorescence detection device for on-site diagnosis according to an embodiment of the present invention.
FIG. 2 is a schematic diagram showing a biochip in the fluorescence detection device for field diagnosis of FIG. 1.
3 is a configuration diagram showing a fluorescence detection device for on-site diagnosis according to another embodiment of the present invention.

The present invention can be applied to various changes and can have various forms, and the embodiments are described in detail in the text. However, this is not intended to limit the present invention to a specific disclosure form, it should be understood to include all modifications, equivalents, and substitutes included in the spirit and scope of the present invention. In describing each drawing, similar reference numerals are used for similar components. Terms such as first and second may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from other components. The terms used in this application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, terms such as “comprises” or “consisting of” are intended to indicate the presence of features, numbers, steps, operations, components, parts or combinations thereof described in the specification, one or more other features. It should be understood that the existence or addition possibilities of fields or numbers, steps, actions, components, parts or combinations thereof are not excluded in advance.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. Terms such as those defined in a commonly used dictionary should be interpreted as having meanings consistent with meanings in the context of related technologies, and should not be interpreted as ideal or excessively formal meanings unless explicitly defined in the present application. Does not.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is a configuration diagram showing a fluorescence detection device for on-site diagnosis according to an embodiment of the present invention.

Referring to FIG. 1, the fluorescence detection device 10 for on-site diagnosis according to the present embodiment includes a light source unit 100, a variable focus lens 200, a mems scanner (300), a scanning lens (400), It includes a biochip (biochip, 500), a temperature control unit 600 and a fluorescence detection unit 700.

 The light source unit 100 irradiates light having a straightness, and the excitation light irradiated from the light source unit 100 is finally delivered to the reaction chambers 550 of the biochip 500. That is, the light generated from the light source unit 100 is formed of constant scanning light and scanned into the biochip 500.

Meanwhile, the light source unit 100 uses a laser as a light source, and the laser light source includes a plurality of lasers irradiating light having different wavelengths.

That is, as shown in FIG. 1, the plurality of lasers may include a first laser 110, a second laser 120, a third laser 130 and a fourth laser 140. In this case, the first to fourth lasers irradiate light having different wavelength values, and may be moved by the first transfer stage 150 and selected as a light source.

By the first transfer stage 150, any one of the lasers is selectively aligned with the variable focus lens 200 so that the laser can be scanned.

On the other hand, although not shown, it is obvious that the number of lasers and the wavelength value of each laser can be variously set.

Before the excitation light irradiated from the light source unit 100 is scanned into the biochip 500, the excitation light is scanned into the MEMS scanner 300. The MEMS scanner 300 includes a reflector that rotates two axes at a certain angle to reflect the incident light, and is composed of a driving unit that drives it, as shown in FIG. 1 to reflect the incident light in various directions. Likewise, rotation is possible around a fixed axis. Accordingly, light incident on the MEMS scanner 300 may be reflected in various directions, and through this, laser light incident linearly may be reflected to the surface light source.

That is, the MEMS scanner 300 may reflect the incident light while changing the angle of reflection of the incident light through rotation around a fixed axis and provide it as a surface light source on the scanning lens 400.

Here, the scanning lens 400 is located between the MEMS scanner 300 and the biochip 500 to convert the light reflected by the MEMS scanner 300 into parallel light, and the light to the biochip 500 Spread to be investigated.

Meanwhile, on the path of light from the light source unit 100 to the MEMS scanner 300, the variable focus lens 200 for adjusting a focal length so that an image of a light source is formed on the biochip 500 is disposed. The variable focus lens 200 serves to focus excitation light irradiated from the light source unit 100, and focuses the excitation light to scan the MEMS scanner 300.

At this time, as shown, when the variable focus lens 200 and the MEMS scanner 300 are not positioned in a line, in this embodiment, the light scanned by the variable focus lens 200 is precisely the MEMS scanner. In order to enter the 300, a focus control mirror 250 may be additionally disposed.

That is, the focus control mirror 250 is located in the direction in which the focused light is traveling in the variable focus lens 200 to reflect the focused light to transmit the focused light to the reflector in the MEMS scanner 300. You can do it.

The excitation light irradiated from the light source unit 100 is applied to the sample contained in the reaction chambers 550 of the biochip 500, and the emitted light is emitted to the fluorescence detection unit 700 emitted from the fluorescent label included in the sample. Check for existence or pattern.

The biochip 500 performs a series of biochemical reactions using injected samples and reagents. The sample may be in various forms, such as nucleic acids or proteins. The nucleic acid may be selected from the group consisting of DNA, RNA, PNA, LNA and hybrids thereof, and the protein may be selected from the group consisting of enzymes, substrates, antigens, antibodies, ligands, aptamers and receptors.

More specifically, referring to FIG. 2, the biochip includes an inlet part 510, a mixing channel 520, a mixing chamber 530, a moving channel 540, a plurality of reaction chambers 550 and an outlet. (Outlet) unit 560.

A plurality of reagents each reacting with a specific bacteria are injected into the inlet part 510, respectively. In this case, reagents injected into the inlet portion 510 may be injected in a mixed state with a sample. Although not illustrated, a micropump for introducing the reagents into the vicinity of the inlet portion 510 is additionally provided. You may.

Of course, even if the reagents and the sample are injected in a mixed state, since sufficient mixing between them is not performed, they must be mixed with each other while passing through the mixing channel 520 and the mixing chamber 530.

Alternatively, the inlet part 510 may be a part in which a sample is injected and a part in which reagents are injected, and the reagents and samples separated from each other are the mixing channel 520 and the mixing chamber 530. It is mixed with each other while going through.

The injected reagents and samples move through the mixing channel 520 and are primarily mixed with each other. In this case, the mixing channel 520 has a meander structure to facilitate mixing of the reagents and the sample.

Thereafter, the reagents and samples flowing through the mixing channel 520 are accommodated in the mixing chamber 530 and secondary to each other by a separate mixer device (not shown) installed inside the mixing chamber 530. Is mixed with.

Then, the reagents move along the moving channel 540 from the mixing chamber 530 in a state sufficiently mixed with the sample. The moving channel 540 is a channel that is fluidly connected to a plurality of reaction chambers 550 that can receive the samples and reagents mixed with each other and amplify the bacteria contained in the sample while confirming the result of the biochemical reaction.

That is, the reaction chambers 550 provide a place to enhance the reactivity by the reagent by amplifying bacteria contained in the sample mixed with the reagent, and to confirm the reaction result from the outside. , As illustrated, it is connected to the moving channel 540 to form a plurality of spaces in which reagents and samples moving along the moving channel 540 can be moved and stored.

The reaction chambers 550, as shown, a plurality of spaces branched from one moving channel 540 are formed, and these spaces are along the extending direction of the moving channel 540, the moving channel 540 ) Can be formed by branching to each side.

Thus, the spaces formed by the reaction chambers form a large area as a whole to correspond to the surface light incident through the MEMS scanner 300, and accordingly, the surface reacts to detect reagents reacted in a large area at a time. can do.

That is, the reagent and the sample are located on the reaction chambers 550, and a target material such as bacteria is amplified, and thus amplified, so that the result of reacting with the reagent can be more easily confirmed from the outside.

For example, the reaction chambers 550 may be configured as a chamber for detecting a positive reaction, a chamber for detecting a negative reaction, and a chamber for detecting a reaction to a sample, in order to detect the presence of a target substance in the sample.

Polymerase chain reaction (PCR) may be performed in the reaction chambers 550. That is, the sample included in the reagents is amplified by passing through the moving channel 540 to be controlled to have a predetermined temperature cycle or maintained at a constant temperature. Accordingly, target substances such as bacteria contained in the sample may be amplified by polymerase chain reaction (PCR) of nucleic acid amplification-based technology.

Thus, the reactivity with the reagents mixed with the sample can be improved, and through this, the reaction result by the reagents can be more easily confirmed externally, in this embodiment.

Further, although the biochip 550 in this embodiment is not shown, in a preparation chamber for preparing a sample from a sample, a liquid storage chamber for storing an enzyme or buffer solution required for analysis of a sample, or a cleaning process in another chamber It may further include a cleaning chamber for storing the required cleaning solution.

That is, in the case of the sample, a separate washing or other process is required before mixing with the reagent, and in this embodiment, it is described that the washing or the like is carried out, so that substances that are unnecessary for the reaction are introduced into the sample. .

The temperature control unit 600 is disposed under the reaction chambers 550 to control the temperature of the reaction chambers 550.

The temperature control unit 600 may be implemented with, for example, a Peltier device. The Peltier element is an electric / thermal conversion element using the Peltier effect. It is a device using a thermoelectric phenomenon in which heat is generated or heat is absorbed at a junction of two metals when two kinds of metals are connected to apply electric current. Therefore, the temperature of the reaction chambers 550 may be increased or decreased by applying a current to the temperature control unit 600 to control the temperature of the reaction chambers.

Alternatively, the temperature control unit 600 is made of a transparent material and is disposed on the surface of the reaction chambers 550 to control the temperature of the reaction chambers 550 and the controller that controls the application of current to the electrodes. It may include, the electrode is electrically connected to the controller by electrical wiring. In this case, the electrode may be made of a transparent material such as a carbon nanotube (CNT: caborn nano tube) or an indium tin oxide (ITO) film.

Furthermore, the temperature control unit 600 may further include a temperature sensor, and may control the temperature of the reaction chambers 550 based on the detection signal of the temperature sensor.

Meanwhile, in order to amplify the sample, the temperature conditions of the reaction chambers 550 are controlled, and at the same time, fluorescence generated by irradiating light to the reaction chambers 550 through the fluorescence detector 700 is detected to detect the sample. The amplification level of can be monitored in real time.

The fluorescence detection unit 700 may detect in real time the emission fluorescence generated by reflection from the reaction chambers 550 where the amplified sample and the reagent are mixed.

As described above, when the laser light of a specific wavelength generated from the light source unit 100 is provided to the reaction chambers 550 of the biochip 500 through the MEMS scanner 300 and the scanning lens 400 , In the reaction chambers 550, when a reagent that reacts with the amplified sample is present, fluorescence is generated according to the reaction between the reagent and the sample, which is detected by the fluorescence detector 700.

Thus, the reagent reacted with the sample may be identified according to the type of fluorescence detected by the fluorescence detector 700, and accordingly, the type of target material such as bacteria reacting with the reagent may be identified. In addition, as shown in FIG. 1, the fluorescence detector 700 may be arranged such that the excitation light irradiated to the sample and the optical path of the generated fluorescence generated therefrom form a predetermined angle, for example, a right angle.

Thus, it is possible to detect fluorescence generated by the reflection of the excitation light applied through the MEMS scanner 300 by the reaction chambers 550 at a more accurate location.

3 is a configuration diagram showing a fluorescence detection device for on-site diagnosis according to another embodiment of the present invention.

The fluorescent detection device 20 according to the present embodiment is illustrated in FIG. 1 except that the temperature control unit 600 includes first and second temperature control units 610 and 620 and a second transfer stage 650. And since it is substantially the same as the fluorescence detection device 10 for on-site diagnosis described with reference to FIG. 2, the same reference numerals are used and redundant descriptions are omitted.

As illustrated in FIG. 3, when the variable focus lens 200 and the MEMS scanner 300 are positioned in a line, light transmitted through the variable focus lens 200 is directly incident to the MEMS scanner 300. Therefore, the focus control mirror 250 described above may be excluded.

Meanwhile, in the present embodiment, the temperature controller 600 includes first and second temperature controllers 610 and 620 and a second transfer stage 650 to control the temperature of the reaction chambers 550. . In this case, the first temperature control unit 610 may increase the temperature of the reaction chambers 550 to a first temperature or a first temperature range, and the second temperature control unit 620 may include the reaction chambers 550 ) May be increased to a second temperature or a second temperature range.

In this case, the first and second temperature controllers 610 and 620 may be selectively positioned below the reaction chambers 550, and for this purpose, the first and second temperature controllers 610, The positions of 620 are switched to each other by the second transfer stage 650.

That is, the first and second temperature control units 610 and 620 select one of the first and second temperature control units 610 and 620 through a position change by the second transfer stage 650. Thus, the reaction chambers 550 may be heated by being located under the reaction chambers 550, thereby raising the temperature within the reaction chambers 550 to a temperature suitable for amplification and progress of the reaction. I can do it.

That is, target materials such as bacteria included in the samples may be various, and accordingly, the optimum temperature for amplification may be different. Accordingly, as in the present embodiment, the first and second temperature controllers 610 and 620 are selectively provided so that the reaction chambers 550 can be heated to different temperatures, which is optimal for amplification. Amplification of the sample can be performed more effectively by providing a temperature corresponding to the temperature of.

In this case, the first temperature and the second temperature, which are the temperatures provided by the first and second temperature control units 610 and 620, may be different from each other, and, alternatively, provided by the first temperature control unit 610 The range of the first temperature and the range of the second temperature provided by the second temperature control unit 620 may be different.

Furthermore, three or more temperature controllers are provided to perform heating at three or more different temperatures, or different temperature ranges, and may be transferred to and selected by the second transport stage 650.

According to the present embodiments, by applying a variable focus lens and a MEMS scanner with low power consumption, and configuring a microfluidic biochip using a small amount of reagent, it is possible to provide a compact detection device suitable for on-site diagnosis.

In addition, by using a variable focus lens, it is possible to adjust the focal length of light incident on the sample surface, and it is possible to provide focused light having a high light intensity on the sample surface.

In addition, since the focused light source is evenly irradiated by the MEMS scanner, there is an advantage that the intensity of light excited over the entire surface of the sample is increased, and accordingly, it is possible to provide focused light without a mechanism that functions as an optical waveguide.

In addition, it is possible to maximize the heat transfer efficiency through a microfluidic biochip using a small amount of reagent, thereby maximizing the polymerization element chain reaction (PCR) and reducing the time.

In this case, in order to perform detection of the target material in a larger area by a light source provided as a surface light source, a plurality of spaces may be formed in the reaction chambers, thereby relatively quickly and easily reacting with various samples. Can be detected.

In addition, considering the difference in amplification temperature according to a sample, by providing a plurality of temperature control units for heating at different temperatures or different temperature ranges, it is possible to provide optimal amplification conditions.

Although described above with reference to the preferred embodiment of the present invention, those skilled in the art may variously modify and change the present invention without departing from the spirit and scope of the present invention as set forth in the claims below. You will understand that you can.

100: light source unit 200: variable focus lens
300: MEMS scanner 400: scanning lens
500: biochip 600: temperature control unit
700: fluorescence detection unit

Claims (12)

A light source unit that irradiates light;
A variable focus lens that focuses light emitted from the light source unit;
A mems scanner that converts the light focused from the variable focus lens into surface light;
A biochip in which a sample mixed with a reagent is amplified and surface light incident through the MEMS scanner is incident;
A light detector for detecting fluorescence reflected from the biochip as the target material of the sample reacts with the reagent; And
It includes a temperature control unit for amplifying the sample in the biochip by heating the temperature of the biochip,
The temperature control unit,
A first temperature control unit that raises the biochip to a first temperature;
A second temperature control unit which raises the biochip to a second temperature different from the first temperature, and is spaced apart from the first temperature control unit; And
And a transfer stage for switching the positions of the first temperature control unit and the second temperature control unit and selectively placing the first temperature control unit and the second temperature control unit under the biochip. . According to claim 1, The light source unit,
A plurality of lasers irradiating laser light of different wavelengths; And
And a first transfer stage for selectively aligning the lasers to the variable focus lens. According to claim 1,
A fluorescence detection device further comprising a focus control mirror positioned in a direction in which the focused light from the variable focus lens travels and reflecting the focused light and transmitting it to the MEMS scanner. According to claim 1,
A fluorescence detection device further comprising a scanning lens that converts and diffuses the surface light incident from the MEMS scanner into parallel light and provides the biochip. delete delete delete According to claim 1, The temperature control unit,
A fluorescence detection device comprising a Peltier element or ITO (Indium Tin Oxide) of a transparent material that generates heat when a current is applied. According to claim 4, The MEMS scanner,
A fluorescence detection device characterized by converting the incident light into surface light reflecting the entirety of the scanning lens while changing the angle of reflection of the incident light through rotation. According to claim 1, The biochip,
An inlet unit for injecting a plurality of reagents mixed with the sample, respectively;
A mixing channel in which the sample and reagents injected through the inlet part are mixed;
A mixing chamber connected to the mixing channel to further mix the sample and reagents;
A moving channel through which the samples and reagents mixed in the mixing chamber move;
A plurality of reaction chambers to which the sample and reagents mixed by branching from the moving channel react; And
And an outlet portion for discharging the sample and reagent reacted in the reaction chambers. The method of claim 10, wherein the reaction chamber,
A fluorescence detection apparatus characterized by forming a plurality of spaces branched from the moving channel, the spaces forming an area corresponding to the surface light incident from the MEMS scanner. The method of claim 10, wherein the reagents,
Located in the reaction chamber, the fluorescent detection device, characterized in that for performing a fluorescence reaction with the amplified target material of the sample.

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