WO2004063731A1 - Optical sensor - Google Patents

Abstract

An optical sensor comprises a semiconductor light source unit for emitting a light with which a labeled substrate of a sample is illuminated, a focusing lens for focusing the light generated when the labeled substance is irradiated with the illuminating light, a filter for selectively transmitting the light generated when the labeled substance is excited by the illuminating light, and an optical sensing unit having an optical sensing element for sensing the light having passed through the focusing lens. The optical path of the illuminating light is different from the optical path of the optical sensing unit.

Description

 Specification

 Photodetector

Related application

 This application was filed on Jan. 16, 2003, filed on January 16, 2003, and was filed on January 23, 2003, and was filed on April 23, 2003. Priority is claimed based on Japanese Patent Application No. 2003-111118344, the contents of which are incorporated into this application.

Technical field

 The present invention relates to a photodetection device applied to an inspection device for a biological substance used for analysis of nucleic acids such as DNA and DNA, antigen-antibody reaction, and protein binding reaction. '' Background technology

 The most commonly used genetic testing methods include extracting nucleic acids from biological samples and using PCR (Polymerase Chain Reaction) or NASBA (Nucleic Acid Sequence-Based Amplification). The target gene is amplified by amplifying the nucleic acid labeled with a radioisotope or a fluorescent dye using the method of amplification (based on J), and the base sequence of the target gene or Methods for measuring the concentration are known.

The electrophoresis method is used for analysis of gene expression and mutation analysis. However, the electrophoresis method has problems such as that it takes time and effort to perform the measurement, and there is a limit to the measurement that can be performed at one time. Therefore, recently, capillary electrophoresis, which allows a fluorescently labeled nucleic acid to react in a plurality of cavities and rapidly process many samples at once, has been widely used. According to the capillary electrophoresis method, the measurement can be performed in a shorter time and more easily than the method using the conventional electrophoresis method.

 In addition, recently, a new testing method using a DNA chip that can perform multiple gene tests simultaneously has been developed. The DNA chip has a large number of DNA probes immobilized on the surface of a glass substrate, and a large number of oligo probes synthesized on a small area on a silicon wafer by applying a semiconductor manufacturing process. There is a DNA chip.

 In the test method using a DNA chip as described above, multiple nucleotide sequences and expression levels of DNA in a sample can be determined simultaneously.Moreover, many gene expression levels / multiple mutations can be determined by applying a DNA chip. It is possible to carry out various tests, such as analysis of data. Furthermore, from data obtained using a DNA chip, many genes can be classified into multiple loops, and information on gene fluctuations associated with development and differentiation can be obtained. However, the gene analysis method using a DNA chip has the following problems. It has the advantage of performing many tests at once, but requires long test times. Since there are many inspection steps throughout and a complicated operation is required, it is difficult to obtain a reproducible detection result.

In order to overcome the above-mentioned problems, as a carrier for a DNA chip, it is similar to a DNA chip in a short time with good reproducibility. A method using a porous filter that can be used for inspection and a method using an electric force to perform the hybridization reaction have been developed (for example, Japanese Patent Application Laid-Open No. 9-50484864). Japanese Patent Publication No. JP-A-2000-501 and Japanese Patent Publication No. JP-A-2001-501.

 In addition, a gene inspection device using a DNA microarray has recently been considered. In this system, genetic analysis is performed using fluorescence emitted from reaction products generated in the reaction chamber of the DNA microarray. This device is a microscope-based device. A hybridization reaction was caused between the target nucleic acid and the nucleic acid probe, which had been fluorescently labeled in advance, in the reaction tank of the DNA microarray, and the reaction was captured by the DNA microarray. A fluorescence signal from a fluorescent substance is acquired, and a fluorescence image is obtained based on the signal. Here, the DNA microarray has a reaction tank (liquid storage section).

 In such a genetic testing method using a DNA microarray, a fluorescent substance is efficiently excited to generate stable fluorescence from a reaction product generated in a reaction chamber of the DNA microarray. This is extremely important in performing highly accurate gene analysis.

 Conventionally, LED (Light Emitting Diode) light source

(Excitation light in this case) is used to irradiate the fluorescent substance with the excitation light to excite the fluorescent substance, and to provide a filter means for selectively transmitting the wavelength of the incident light with respect to the fluorescence emitted by the excited fluorescent substance. Through An apparatus has been proposed in which a fluorescent signal is received by a body imaging device and a fluorescent image is obtained based on the fluorescent signal (for example, see Japanese Patent Application Laid-Open No. 10-132744). .

 The light from the LED and the like is irradiated to the well as excitation light via a dichroic mirror and an objective lens, and the fluorescent light emitted from the sample is passed through the objective lens and the dichroic mirror. An apparatus has also been proposed in which a photomultiplier tube is used for detection via a pinhole cutout (see Japanese Patent Application Laid-Open No. 200-11616).

 A device in which semiconductor light sources are two-dimensionally arranged to illuminate a sample as a surface light source through a capacitor lens, and light transmitted through the sample is made incident on an objective lens, that is, 2 Illumination light from a three-dimensionally arranged semiconductor light source is sent along the optical axis, and is incident on the objective lens while being aligned with the optical axis of the objective lens, so that an enlarged image can be obtained. An apparatus has been proposed (see Japanese Patent Publication No. 7-122628).

 Using a semiconductor excitation light source, irradiate the sample with light through the objective lens to excite the fluorescent dye inside the sample, and detect the fluorescence from the sample through the same optical system as the incident light with a photodetector. A device that performs this operation has also been proposed (see No. 113-615 15 282).

In general, LED light sources do not have sufficient light intensity to excite the fluorescent material and cannot generate sufficient fluorescence from the fluorescent material. For this reason, when an LED is used as a light source, it is often difficult to receive light with a commonly used photodetector such as a CCD camera. Therefore, when an LED light source is used as a light source for exciting a fluorescent substance, the output intensity should be as low as possible. In addition to selecting a model with a large surface area, bundle two or more LEDs so that they are as close to the sample surface as possible, and make sure that a sufficient amount of excitation light is uniformly applied to the fluorescent material. It is necessary to take measures such as doing so.

Disclosure of the invention

 According to one aspect of the present invention, there is provided a photodetector capable of efficiently irradiating sufficient light with high efficiency and performing highly reliable fluorescence detection.

 A light detection device according to one aspect of the present invention includes: a semiconductor light source unit that emits light for irradiating a labeled substance of a sample; and a device that collects light emitted by irradiating the labeled substance with the irradiation light. A condenser lens for emitting light, a filter for selectively transmitting light emitted when the labeling substance is irradiated with light by the irradiation light, and a light for transmitting light passing through the condenser lens. And a photodetector having a photodetector for detection, wherein the optical path of the excitation light is different from the optical path of the photodetector.

 A light detection device according to another aspect of the present invention includes: a semiconductor light source unit that emits light for irradiating a labeled substance of a sample; and the semiconductor light source unit includes a semiconductor light emitting element and an optical element. A condenser lens for condensing light emitted by irradiating the labeling substance with the irradiation light, and selectively condensing light emitted by irradiating the labeling substance with the irradiation light. And a photodetector having a photodetector that detects light passing through the light-collecting lens.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a schematic diagram of an optical inspection apparatus according to the first embodiment of the present invention. The figure which shows schematic structure.

 FIG. 2 is a diagram showing a schematic configuration of an example of a DNA reaction vessel used in the first embodiment.

 FIG. 3 is a diagram showing a schematic configuration of another example of the DNA reaction vessel used in the first embodiment.

 FIGS. 4A and 4B are views for explaining light irradiation on a sample according to the first embodiment.

 FIG. 5 is a block diagram showing the entire optical inspection apparatus according to the first embodiment.

 FIG. 6 is a diagram showing a schematic configuration of an LED light source unit according to a first modification of the first embodiment.

 FIG. 7 is a diagram showing a schematic configuration of an LED light source unit according to a second modification of the first embodiment.

 FIG. 8 is a diagram showing a schematic configuration of an LED light source unit according to a third modification of the first embodiment.

 FIG. 9 is a diagram showing a schematic configuration of a main part of an optical inspection device according to a second embodiment of the present invention.

 FIG. 10 is a view showing a schematic configuration of an optical inspection device according to a modification of the second embodiment.

 FIGS. 11A and 11B are diagrams showing a schematic configuration of an optical inspection device according to a third embodiment of the present invention.

 FIG. 12 is a diagram showing a schematic configuration of a measurement optical system used in the third embodiment.

FIG. 13 is a block diagram showing an LED light source unit drive circuit used in the third embodiment. FIG. 14 is a diagram illustrating a schematic configuration of an LED light source driving circuit used in the third embodiment.

 FIG. 15 is a diagram showing a schematic configuration of an optical inspection device according to a fourth embodiment of the present invention.

 FIG. 16 is a diagram showing a schematic configuration of an optical inspection device according to a first modification of the fourth embodiment.

 FIG. 17 is a diagram illustrating a schematic configuration of an optical inspection device according to a second modification of the fourth embodiment.

 FIG. 18 is a diagram showing a schematic configuration of an optical inspection device according to a fifth embodiment of the present invention.

 FIG. 19 is a diagram illustrating a schematic configuration of an optical inspection device according to a modification of the fifth embodiment.

BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, embodiments of the present invention will be described with reference to the drawings. In the following embodiment, a description will be given of a fluorescence detection apparatus that irradiates a fluorescent substance with excitation light. A fluorescent detector that inspects fluorescence from the fluorescent substance will be described. It is of course applicable to a detection device that detects light or reflected light (hereinafter, these are collectively referred to as a “light detection device”). Various fluorescent dyes and fluorescent glass particles can also be used as the fluorescent substance. In addition, metal particles or dielectric particles are used as a labeling substance when detecting with scattered light or reflected light. For example, in addition to metal particles, fine particles such as silver, platinum, and silicon, and latex particles can be used. In particular, fine particles of metals such as gold, silver, and platinum have a particle size of 10 to 100 nm. However, it is particularly preferred because the speed of the particles in motion is optimal. Latex particles having a particle size of 0.1 to 1 m are also particularly preferred because the speed of the particles in motion is also optimal. The appropriate particle size is determined by the specific gravity of the particles and the speed of the browning motion. Here, the motion state of the particles includes, for example, Brownian motion and vibration.

 As used herein, the term “specific binding substance” refers to hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, RNA, and PNA. And the like, meaning a substance that can specifically bind to a biological substance, and is called a probe.

 A “biological substance” is a substance that specifically binds to a known specific binding substance placed at a predetermined position on a substrate on which a probe is immobilized, and is extracted and isolated from a living body. The term refers to substances that have been subjected to chemical treatment, chemical treatment, chemical modification, and the like, as well as substances directly extracted from living organisms. For example, substances such as hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, RNA, and PNA.

"Specific binding between a biological substance" and a "specific binding substance" means that an unstable double strand is formed between complementary nucleotide sequences found in DNA, RNA, etc. Or highly specific binding, such as in the case of hybridization (hybridization) or selectively reacting only with a specific substance, such as an antigen and an antibody or a biotin and avidin. (First Embodiment)

 FIG. 1 is a diagram showing a schematic configuration of an optical inspection device according to a first embodiment of the present invention. An optical inspection device according to each embodiment of the present invention includes a light-collecting lens, a sample holding unit, a light source, and a photodetector.

 In FIG. 1, the optical inspection apparatus includes a sample stage 1, and a DNA reaction container 2 as a subject is placed on the sample stage 1.

 FIG. 2 is a diagram showing a DNA slide glass reaction vessel 3 as a specific example of the DNA reaction vessel 2. The DNA slide glass reaction vessel 3 has a sample tank 301 arranged in a slide glass-like reaction vessel. In the sample tank 301, a DNA microarray 302 is laid.

 In the above configuration, when a sample solution (test sample) is injected into the sample tank 301, a hybrid is formed between the nucleic acid probe and the target nucleic acid previously fluorescently labeled in the DNA microarray 302. Redidation reaction occurs. By this hybridization reaction, a fluorescent substance, for example, a fluorescent dye substance, captured by the DNA microarray emits fluorescence. In this case, the sample solution that did not contribute to the reaction is washed together with the buffer.

FIG. 3 is a diagram showing a specific example of another DNA reaction container 4. The DNA reaction container 4 is formed of a plastic material. A plurality of sample vessels 401 are arranged in the DNA reaction vessel 4, and a DNA microarray 402 is laid in the sample vessel 401. In the above configuration, when the sample solution (test sample) is injected into the sample tank 401, the DNA microarray 4 In step 02, a hybridization reaction occurs between the target nucleic acid and the nucleic acid probe that have been fluorescently labeled in advance. A fluorescent substance, for example, a fluorescent dye, captured in the DNA microarray by the hybridization reaction emits fluorescence. In this case, the sample solution that did not contribute to the reaction is washed together with the buffer.

 Above the sample stage 1, an objective lens 5 is arranged as a focusing lens. The objective lens 5 is positioned on a perpendicular line from one surface of the sample stage so that its optical axis 5a coincides with the center of the sample vessel 201.

 The objective lens 5 is held by an objective lens holding mechanism 6. The objective lens holding mechanism 6 has a cylindrical holding member 600. The base end of the objective lens 5 is fitted inside the holding member 600. A plurality of screw holes 62 are formed on the peripheral surface of the holding member 61 at equal intervals along the circumferential direction. A fixing member 603 is provided so that the peripheral surface of the holding member 601 abuts. The fixing member 603 is fixed to a device main body (not shown), and an elongated hole 604 along the optical axis direction of the objective lens 5 is formed in the fixing member 603. A position adjusting screw 605 is screwed into the screw hole 602 on the peripheral surface of the holding member 601 via the elongated hole 604. By loosening the screws for position adjustment 605, the objective lens 5 moves along with the holding member 601 along the elongated hole 604 in the direction of the optical axis 5a. This allows you to adjust the focus.

On the optical axis 5 a of the objective lens 5, a filter 7, an imaging lens 8, and a light detection device that together with the objective lens 5 constitute a light detection unit. Container 9 is placed. The filter 7 selectively transmits the fluorescence emitted when the fluorescent substance labeled on the sample 202 is excited. The imaging lens 8 forms the fluorescent light selected by the filter 7 on the detection surface of the photodetector 9. The photodetector 9 detects the intensity of the fluorescence collected on the detection surface, converts the detected intensity into an electric signal, and outputs the electric signal to a computer 10 described later. Here, the imaging lens 8 may be a glass lens such as BK7 used for a normal lens as a material, but the imaging lens 8 may be quartz glass, a plastic lens, or Diffractive optical elements, liquid crystal lenses, etc. Elements and materials capable of condensing ordinary visible light can be used.

 Around the objective lens 5, a plurality of LED light source units 11 (for example, four LED light source units 11; in FIG. 1, three LED light source units 11) are provided as semiconductor light source means. (Shown) is arranged.

 The LED light source unit 11 has a cylindrical LED light source holder 111. The LED light source holder 1101 has an LED light source 1102 disposed therein. On the optical path of the light emitted from the LED light source 1102 disposed inside the LED light source holder 1101, a non-pass filter 1103 is disposed. The non-linear filter 1103 mainly passes near the wavelength range of the light emitted from the LED light source 1102.

 The spectral characteristics of the emission wavelengths of the LED light sources 1102 included in the plural LED light source units 11 are the same.

The light from the LED light source unit 11 is supplied to the sample tank on the sample stage 1. The 201 sample 202 is irradiated as excitation light. In this case, it is necessary to mount each LED light source unit 11 for the sample 202 at a position that illuminates the sample 202 surface with uniform brightness. That is, excitation unevenness does not occur on the sample 202 surface due to the incident angle of the light irradiated on the sample 202, and the sample 202 having a three-dimensional portion on part or the entire surface is also shadowed. Each LED light source unit 11 is mounted so that no excitation mura occurs.

 Specifically, as shown in FIGS. 4A and 4B, the central axis of the light of each LED light source unit 11 (total of four light sources) corresponds to the periphery a, b, c of the sample 202 surface. Through each of the LED light source units 11 so that they intersect at a point e (or f) at a spatial position vertically above or below the approximate center axis of the sample 202 surface. The mounting position is set. In this case, if it is possible to prevent the occurrence of excitation mura on the sample 202 surface, even if the position is off the center axis of the sample 202 surface, It is also possible to set the mounting position of each LED light unit 11 so that it intersects at one point of the upper or lower spatial position.

 The LED light source unit 11 moves the sample 20

The excitation light applied to the sample 2 is not limited to the sample

0 2 It is also conceivable to irradiate the surface perpendicular to the optical axis of the objective lens 5 inside.

 The main part of the optical inspection device thus configured is installed in the light-shielding box 12 and is shielded from the outside.

Figure 5 shows a block diagram of the entire optical inspection system. In FIG. 5, the computer 10 has a monitor 13 as a display means. The computer 10 includes a fluorescence detection unit 14 including a main part of the apparatus shown in Fig. 1 and an LED light source unit driving circuit as a driving means for driving each LED light unit 11. 1 5 and are connected. When receiving a control command from the computer 10, the LED light source drive circuit 15 is driven by one of the LED light source units 11 in the fluorescence detection unit 14. At the same time as the unit 11 is determined, the drive current of the LED light source unit 11 is set and the drive current is supplied to the LED light unit 11.

 The operation of the embodiment configured as described above will be described.

 Now, when the computer 10 sends a control command to the LED light source unit drive circuit 15, the LED light source unit drive circuit 15 responds to the content of the command at this time, and the fluorescence detection unit 1 4 Determine the LED light source unit 11 driven from the power of the LED light source unit 11 in 1 and at the same time determine the magnitude of the drive current of the LED light source unit 11 Supply drive current to unit 11.

 Here, in the following description, it is assumed that all (four) LED light source units 11 in the fluorescence detection unit 14 are simultaneously driven.

 When light is emitted from the LED light source 1102, light of a predetermined wavelength range that has passed through the node pass filter 1103 is sent to the sample 202 in the sample tank 201 on the sample stage 1. The light is emitted as excitation light.

In this case, the light of each LED light source 1 102 As shown in Fig. 4A and Fig. 4B, the central axis of each light passes through the periphery a, b, c, d of the specimen 202 surface so that excitation unevenness does not occur on the surface. The LED light sources 1102 are arranged so as to intersect at one point e (or ί) of a spatial position vertically above (or below) the substantially central axis of the sample 202 surface.

 When the LED light source 1102 irradiates the sample 202 with excitation light, the fluorescent substance in the sample 202 emits fluorescence. The fluorescent light is condensed by the objective lens 5, passes through the filter 7 and the imaging lens 8, and forms an image on the detection surface of the photodetector 9. The photodetector 9 detects the intensity of the fluorescent light, converts it into an electric signal, and outputs the electric signal to the computer 10.

 The computer 10 performs image processing and signal analysis such as contour enhancement, contrast correction, color correction, etc., and displays it on the monitor 13 as a fluorescent image.

 As described above, in the first embodiment, a plurality of LED light source units 11 (for example, four LED light source units) having the LED light sources 1102 having the same emission wavelength spectrum are used. The unit 1 1) was arranged side by side around the objective lens 5, and the excitation light was irradiated on the sample 202 simultaneously by the multiple LED light source units 1 1. Excitation light of sufficient intensity to excite the fluorescent substance in 2 can be efficiently obtained.

Also, the center axis of each light passes through the periphery of the sample 202 surface and the sample 202 so that the light from the plurality of LED light sources 1102 becomes uniform on the sample 202 surface. Since they are set to intersect at one point vertically above or below the space on the approximate center axis of the two surfaces, The shadow and excitation unevenness on the surface of the sample 202 can be eliminated, and more stable fluorescence can be generated than the fluorescent substance generated in the sample tank 201.

 In addition, since the light from the LED light source 1102 is radiated toward the surface of the sample 202 obliquely from the optical axis of the objective lens 5 at a predetermined angle, the sample 202 Specularly reflected light from the surface does not pass through the light receiving optical path of the objective lens 5 as it is and enters the photodetector 9 as noise light, so that noise light can be reduced. You.

 Furthermore, since the LED is used as an excitation light source for exciting a fluorescent substance, it is inexpensive, has a long life, generates little heat, and is safe. In addition, power consumption can be reduced. In addition, the device configuration can be made small and highly portable.

 Furthermore, since the LED light source unit 11 has only the band-pass filter 1103 as an optical element other than the LED light source 1102, the configuration is simple. Not only is it easy to assemble and adjust the optical axis, but it is also useful because it leads to cost reduction.

In addition, by disposing the LED light source unit 11 around the objective lens 5, the optical path of the excitation light and the optical path having the objective lens 5 for fluorescence detection are separated from each other. There is no need to install extra optical elements such as a Crook Mirror, and a simple device configuration can be achieved in this regard. In addition, since there is no optical element such as a die-cloth mirror, reflected light and scattered light generated on the surface of these optical elements, and light passes through these optical elements As a result, the loss of the light intensity detected by the photodetector 9 can be minimized because the fluorescence intensity does not attenuate. Furthermore, since the excitation light path through which the excitation light passes and the light path of the fluorescence detection means are completely separated, adjustment for aligning both optical axes is unnecessary, and the adjustment of the optical axis of the entire apparatus can be simplified. . Therefore, from these facts, highly reliable fluorescence detection can always be performed.

 (First modification)

 As the above-described LED light source unit 11, for example, an LED light source unit 16 as shown in FIG. 6 can be used. FIG. 6 shows a first embodiment of the first embodiment. FIG. 9 is a diagram showing a schematic configuration of an LED light source unit 16 according to a modification.

The LED light source unit 16 has a cylindrical LED light source holder 1601. The LED light source holder 1601 has an LED light source 1602 disposed therein. In addition, on the optical path of the light emitted from the LED light source 1602 disposed inside the LED light source holder 16-011, a non-pass filter 1603, a diffusion plate 1604, A light lens 1605 is arranged. Here, the Nordpass filter 1603 mainly transmits near the wavelength range of the light emitted from the LED light source 1602. The diffuser plate 164 is used to suppress the non-uniformity (intensity unevenness) of the light intensity from the LED light source 1602 that has passed through the non-pass finoletor 1603. As the diffusion plate 1604, for example, "frosted glass" or a translucent plastic plate is used. The condenser lens 1605 is the light diffused by the diffuser 1604. And focuses the diffused light at a position determined by the focal length. In this case, the positions of the non-pass filter 1603 and the diffuser 1604 are the same as the diffuser 1604, the nozzle 1601 and the condenser lens. The order may be 1 6 0 5.

The LED light source unit 16 according to the first modified example has a condensing lens 1605 on the optical path of light emitted from the LED light source 1602. It can irradiate a focused beam. Thus, for example, if a large condensing lens 1605 having a NA of 0.95 or more is used, a very small area on the sample 202 surface, for example, a diameter of 0.5. Light can be collected in the range of about μm, and a specific extremely narrow range of excitation can be achieved. In addition, since the excitation light from the LED light source 1602 becomes a condensed beam and the area of the irradiation cross section is small, a portion other than the sample 202 surface, such as the side wall of the sample tank 201, is used. Thus, it is possible to prevent the excitation light from irradiating the sample with the excitation light and generating the noise light, which may be a factor of mixing with the fluorescence from the sample 202. Further, if the direction of each LED light source 1602 is adjusted so that the excitation light from the LED light source 1602 is focused on one point in the plane of the sample 202, the LED light source 16 It is possible to irradiate the excitation light from O2 only to a desired specific portion in the surface of the sample 202. As a result, it is possible to efficiently irradiate the excitation light from the LED light source 1602 to a desired portion in the plane of the sample 202. In addition, since the excitation light from the LED light source 1602 is collected, it is possible to efficiently excite the fluorescent substance. In addition, since light is not diffused, a portion in the sample tank 201 other than the sample 202 surface, for example, a side wall Erroneous irradiation of light can be prevented. (Second modification)

And an LED light source uni Tsu sheet 1 1 described above, for example, _c 7 that can have that you use an LED light source Interview two Tsu sheet 1 7 Do you by 7 is a first embodiment FIG. 14 is a diagram showing a schematic configuration of an LED light source unit 17 according to a second modification.

 The LED light source unit 17 has a cylindrical LED light source holder 1701. The LED light source holder 1701 has an LED light source 1702 disposed therein. In addition, on the optical path of the light emitted from the LED light source 1702 disposed inside the LED light source holder 1701, a bandpass filter 1701, a diffusion plate 1704, and a There is a lens 1705. Here, Band No. The finoletor 1703 and the diffuser 1704 are the same as those described with reference to FIG. The collimating lens 1705 converts the light diffused by the diffusion plate 1704 into parallel light, and emits collimated light. In this case. The positions of the band-noise filter 1703 and the diffuser 1704 are the same as those of the diffuser 1704, the non-pass filter 170, and the coil. The order of the lenses 1705 can be used.

The LED light source unit 17 according to the second modified example has a collimated lens 105 disposed on the optical path of light emitted from the LED light source 1702, so that the sample It is possible to irradiate the flat collimated light onto the 202 surface. This makes it easy to identify the irradiation position in the plane of the sample 202, making it easy to program a program for automatically adjusting the irradiation position by computer control. In addition to this, it is easy and useful to predict the irradiation surface position and its illuminance. Furthermore, even when the position for irradiating the excitation light is manually set, the excitation light can be accurately operated at a desired irradiation position on the sample 202 surface. . In addition, since light is not diffused, it is possible to prevent erroneous irradiation of light in a portion of the sample tank 201 other than the sample 202 surface, for example, including a side wall. it can.

(Third Modification). As a LED light source unit 1 1 described above, for example, _c 8 that can have that you use an LED light source Interview two Tsu sheet 1 8 Do you by shown in FIG. 8, FIG. 14 is a diagram illustrating a schematic configuration of an LED light source unit 18 according to a third modification of the first embodiment.

 The LED light source unit 18 has a cylindrical LED light source holder 1801. The LED light source holder 1801 has an LED light source 1802 disposed therein. In addition, on the optical path of the light emitted from the LED light source 1802 disposed inside the LED light source holder 1801, a non-pass filter 1803 and a diffusion plate 1804 are disposed. ing. Here, the band pass filter 1803 and the diffusion 扳 1804 are the same as those described with reference to FIG. In this case, the position of the non-pass filter 1803 and the diffusion plate 1804 may be in the order of the diffusion plate 1804 and the bandpass filter 1803.

The LED light source unit 18 according to the third modification has the diffusion plate 1804 arranged on the optical path of the light emitted from the LED light source 1802. Irradiate diffuse light without unevenness You can do it. As a result, it is possible to irradiate almost uniformly the excitation light over the entire surface of the sample 202 without unevenness, and it is possible to receive stable and excellent fluorescence with excellent reproducibility. .

 Note that these LED light source units 11, 16, 16, 17 can be used properly according to the size and structure of the sample 202 and the sample tank 201. That is, the LED light source unit 1 1

Each of the LED light sources unit 16, 17, and 18 is composed of an LED light source and an optical element such as a non- ^zero filter, a lens, and a diffusion plate. The excitation light can be condensed, collimated, or diffused in units of light. This makes it possible to control the beam pattern of the excitation light to the sample 202 surface in various ways, and the sample tank 201 of various shapes has more unevenness. It can respond to a certain sample. Also, as the LED light source unit 16 (17), it can be used according to the application. Either the filter 1603 (1703) or the diffusion plate 1604 (1704) may be used, or neither of them may be used. In addition, LED light source units 11, 16,

17 and 18 nodes. Scanners 1103, 1603, 1703, and 1803 are different depending on the spectrum of the emission wavelength of the fluorescent dye used as the sample. ° Needless to say, it may be a filter or a low-pass filter. The condenser lens 1705 6 The collimating lens 1705 may be a glass lens such as BK7 used for ordinary lenses as a material, but the focusing lens 16 0 5 ゃ Collimate lens 1 7 0 5 is quartz glass or plastic lens or diffractive optical element Elements and materials that can condense ordinary visible light, such as liquid crystal lenses, can be used. In the first embodiment, the case where a plurality of LED light source units 11 are arranged around the objective lens 5 has been described. In 1, only one LED light source unit 11 disposed around the objective lens 5 may be used. Even in this case, the same effect as described above can be expected.

 (Second embodiment)

 The second embodiment is an embodiment in which the angle of light irradiation can be adjusted by changing the inclination angle of the LED light source unit with respect to the sample surface.

 FIG. 9 is a diagram illustrating a schematic configuration of the second embodiment. In FIG. 9, the objective lens 21 is held by an objective lens holding mechanism 22. The objective lens holding mechanism 22 has a cylindrical holding member 222. The objective lens 21 is fitted inside the holding member 222. Around the holding member 222, an LED light source unit holder 23 is provided as a semiconductor light source means holding member.

The LED light source unit holder 23 is provided with a plurality of (for example, about 8 to 12) LED light source unit storage holes 2301 at even intervals along the circumferential direction. . These LED light source cut-out holes 2301 are arranged to be inclined toward the surface of the sample 202 on the sample stage 1. The tilt angle is about 45 ° to 60 ° with respect to the optical axis 21a of the objective lens 21 with respect to the axis. In addition, the LED light source unit holder 23 has an L A screw hole 2302 penetrating the ED light source unit storage hole 2301 is provided. The two screw holes 2302 are formed at predetermined intervals in the direction along each LED light source unit accommodation hole 2301.

 The LED light source unit 24 is accommodated in the LED light source unit accommodating hole 2301. The LED light source unit 24 has a cylindrical holoreder 2401, and the LED illuminator 2402 is disposed inside the holoreder 2401. Then, such an LED light source unit 24 is housed in the LED light source unit housing hole 2301 via an O-ring 25. In this case, the O-ring 25 is located between the two screw holes 2302.

 The LED light source unit storage hole 2301 has a bandpass filter 240 on the optical path of the light emitted from the LED light source 2402 of the LED light source unit 24. 3 is located. Here, the non-phosphorescent light source 2403 mainly passes near the wavelength range of light emitted from the LED light source 2402 power.

 In addition, a spacer 26 is disposed between the hologram 2404 of the LED light source unit 24 and the finolators 2403 of the LED light source unit. The spacer 26 is composed of an LED light source 2402 and a band laser. Performs positioning of the fin locator 2403. As a material of the spacer 26, a metal such as aluminum or brass or a plastic is used.

In two screw holes 2302 passing through each LED light source cutout accommodation hole 2301, a tilting screw 27 as a position adjusting means is screwed. Fan screw 27 is for LED light source unit 2 Press the two points across the O-ring 25 on the side of the holder 2 4 0 1. The light irradiation angle can be adjusted by changing the tilt angle of the LED light source unit 24 with respect to the sample 202 surface by adjusting the pressing force of the lifting screw 27 according to the screwing amount. . In this case, the irradiation angle of the light emitted from the LED light source 2402 adjusted by these two tilting screws 27 is ± 2 to 3. It is on the order.

 On the other hand, a plurality of screw holes 222 are formed on the peripheral surface of the holding member 222 at even intervals along the circumferential direction. Further, a fixing member 222 is provided so that the peripheral surface of the holding member 222 contacts. The fixing member 2203 is fixed to the apparatus main body 28 side, and has an elongated hole '224' formed along the optical axis direction of the objective lens 21. Then, a positioning screw 222 is screwed into the screw hole 222 on the peripheral surface of the holding member 222 through the elongated hole 222. As a result, by loosening the screws of the position adjustment screws 222, the objective lens 21 is moved along with the holding members 222 along the elongated holes 222. Move in the optical axis 21a direction. This allows the focus to be adjusted.

 In the above configuration, first, the LED light sources 2402 of each LED light source unit 24 are turned on, and the excitation light emitted from these LED light sources 2402 is applied to the sample 202 surface on the sample stage 1. In this case, the light emission spectrum of the light emitted from the LED light source 2402 is all the same.

In this state, loosen the position adjusting screw 2205 of the objective lens holding mechanism 22 and set the objective lens 21 together with the holding member 222. Focus adjustment is performed by moving the optical axis in the 2 la direction (the direction of arrow A in the figure).

 Next, loosen the screw 27 of the LED light source unit holder 23, and move the LED light source unit 24 in the optical axis direction of the LED light source 2402 (the direction of arrow B in the drawing). Thus, the position of the LED light source unit holder 23 in the height direction is adjusted, and the brightness on the sample 202 surface is adjusted. Subsequently, by adjusting the amount of screwing in by the tilting screw 27 to adjust the inclination angle (in the direction of arrow C in the figure) of the LED light source unit 24 with respect to the surface of the sample 202, the sample 202 was obtained. Fine adjustment of the light irradiation position

 In this way, the brightness of the light on the sample 202 surface is adjusted and the light irradiation angle is finely adjusted, as described in FIGS. 4A and 4B. The central axis of the light emitted from each LED light source unit 24 passes through the periphery of the sample 202 surface and is vertically above or below the substantially central axis of the sample 202 surface. Set to intersect at one point of the position. This makes it possible to illuminate the sample 202 surface uniformly and uniformly.

 In addition, by doing so, it is possible to condense light to a desired specific portion of the sample 202 surface, so that efficient excitation of the fluorescent substance can be performed, and In addition, the excitation light hits the side wall of the sample tank 201 and the like, and it can be suppressed as much as possible the noise light.

In the second embodiment, the screwing amount of the tilting screw 27 is manually adjusted. For example, the screwing of the tilting screw 27 is performed. It is also possible to use a motor at the position corresponding to the adjustment of the control amount, and to operate this motor automatically by linking it with the computer. In addition, the LED light source unit 24 may be configured to be capable of adjusting the position in the height direction and adjusting at least one of the inclination angles.

 (Modification)

 However, in order to illuminate the sample surface uniformly and uniformly with light, it is necessary to check this condition in advance.

 FIG. 10 is a diagram showing a schematic configuration of an optical inspection device according to a modification of the second embodiment, and the same parts as those in FIG. 1 are denoted by the same reference numerals. In this modification, the block diagram of the optical inspection device described with reference to FIG. 5 is used.

 In this modification, a light detector 31 is disposed on the sample table 1 to which light from the plurality of LED light source units 11 is irradiated, instead of the sample tank 201 as light intensity detecting means. Have been. Here, a solid-state image sensor (CCD camera or CMOS sensor), an image pickup tube, or the like is used as the light detector 31.

 The photodetector 31 individually detects the intensity of light from each LED light source unit 11. The detection output of the photodetector 31 is taken into the computer 10 described in FIG.

The computer 10 analyzes the variation of the light intensity from the output of the photodetector 31 corresponding to the intensity of the light from each LED light source unit 11 and outputs the LED light source unit drive circuit 15. And individually controls the current supplied to the LED light source 1102 of each LED light unit 11. In this modification, the magnitude of the current supplied to each LED light source 1102 can be controlled based on the light intensity obtained near the sample surface. This makes it possible to illuminate the specimen surface with uniform brightness. As a result, illumination unevenness on the sample 202 surface can be suppressed, the fluorescent substance in the sample tank 201 can be excited almost uniformly, and highly reliable fluorescence detection can be performed. And can be.

 In this modified example, the magnitude of the current supplied to each LED light source 1102 was adjusted using the computer 10, but each LED light source unit 11 obtained by the photodetector 31 was adjusted. The LED light source unit drive circuit 15 is manually adjusted based on the intensity of light from the LED light source 1102 to adjust the light from the LED light source 1102 to the same brightness. The photodetector 31 may be removed, and the sample tank 201 may be re-installed at this position.

On the other hand, an imaging means (for example, a CCD camera or a CMOS sensor (both not shown)) is used in place of the photodetector 9, and the sample tank 201 is placed on the sample table 1 as it is, and The light from the LED light source 111 of the LED light source unit 11 is individually radiated to the sample tank 201, and the fluorescent light from the sample tank 201 is objective lens 5, filter 7, imaging lens 8 , Each of which is taken by the image pickup means, guided to the computer 10 and displayed on the monitor 13, and the brightness of each pixel of the fluorescent image on the monitor 13 is set to the computer 10. By analyzing the light intensity from each LED light source 1102, the variation in the light intensity is determined, and based on the result, the LED light source unit drive circuit 15 is adjusted, and each of the LED light source unit drive circuits 15 is adjusted. The current supplied to the LED light sources 11.02 can be individually controlled.

 Even in this case, the light from each LED light source 1102 can be adjusted to the same brightness, so that uneven illumination on the sample 202 surface can be suppressed, and the sample tank can be controlled. The fluorescent substance in 201 can be excited almost uniformly.

 (Third embodiment)

 The third embodiment is an embodiment in which a plurality of types of fluorescent dyes are excited using two or more LED light sources having different peak emission wavelengths, and a fluorescent signal can be detected.

 FIGS. 11A and 11B are diagrams showing a schematic configuration of the third embodiment, and the same parts as those in FIG. 9 are denoted by the same reference numerals. In the third embodiment, the block diagram of the optical inspection apparatus described with reference to FIG. 5 is used.

 The first to third LED light source units 41, 42, and 43 having three types of LED light sources having different peak emission wavelengths are mounted on the LED light source unit holder 23. In general, the emission spectrum of the LED light source has a mountain-shaped structure, and has one peak wavelength. In this case, for example, the first LED light source unit 41 uses an LED light source having a light emission peak wavelength of 490 nm, and the second LED light source unit 42 uses the light emission peak wavelength. An LED light source having a wavelength of 52 O nm is used, and an LED light source having a peak emission wavelength of 63 O nm is used as the third LED light source unit 43.

The first to third LED light source units 41, 42, and 43 are L Four ED light source unit holders 23 are arranged symmetrically around the objective lens 21 along the periphery. In other words, these first to third LED light source units 41, 42, and 43 have LED light sources having different emission wavelength spectral characteristics, respectively, and these different spectra The first to third LED light source units 41, 42, and 43 having the same characteristics are arranged at regular intervals along the periphery of the LED light source unit holder 23. Here, the first LED light source unit 41, the second LED light source unit 41, and the third LED light source unit 43 are repeatedly arranged in this order. 'The first to third LED light source units 41, 42, and 43 have the same spectral characteristics at positions facing each other across the objective lens 21. It is arranged as follows.

The first to third LED light source units 41, 42, and 43 are mounted in front of the respective LED light sources (in the drawing, the first LED light source unit). 4) The LED light source 4101 in front of the LED light source 4101 (only the filter 4102 is shown) transmits the light near the peak wavelength of the emission of each LED light source best. _t that is made as having the property of, Roh down de path full I filter 4 1 0 2 of the first LED light source Interview two Tsu DOO 4 1, the wavelength to be the most good rather permeation 4 9 O n ra The second LED light source unit is set to a nearby position. The filter has the best transmission wavelength set around 520 nm, and the third LED light source unit 43 has a non-pass filter with the best transmission wavelength around 630 nm. Set Reply

 Fig. 12 shows the schematic configuration of the photodetection optical system of the device configured as described above. Here, the LED light source unit having the above three different wavelengths at the peak emission wavelength is shown. 4 1, 4 2,

Figure 4 shows the case where the sample is irradiated with excitation light. In FIG. 12, two dichroic mirrors 45 and 46 are arranged on the optical axis above the objective lens 21.

 The dichroic mirror 45 is set so that the reflected light travels in a direction of approximately 45 ° with respect to the optical axis of the objective lens 21, and the peak emission of the fluorescent dye FITC is performed. A transflective spectral characteristic that reflects light having a wavelength slightly longer than the wavelength of 520 nm, for example, light having a wavelength of 550 nm or less, and transmits light having a wavelength longer than that. have. The dichroic mirror 46 is installed so that the reflected light travels in a direction of approximately 45 ° with respect to the optical axis of the objective lens 21, and the fluorescent dye Cy 3 Peak emission wavelength of 5 6

It has a transflective spectral characteristic that reflects light having a wavelength slightly longer than 5 nm, for example, light having a wavelength of 62 nm or less, and transmits light having a wavelength longer than that. I have.

 Further, a CCD camera 48 is disposed on the reflection optical path of the die mirror 45 through a condenser lens 47. In addition, a CCD camera 50 is arranged via a condenser lens 49 in the reflected light path of the die-cloth mirror 46, and a condensing lens 51 is arranged in the transmitted light path. CCD camera 52

The outputs of the CCD cameras 48, 50, 52 are sent to the computer 10. In the above configuration, three types of target DNA are now set for one sample, and different fluorescent dyes, FITC (Forescein-isothiocyanate), Cy3, and Cy5, are used for each of them. Label. Next, a sample solution containing DNA labeled with these fluorescent dyes is dropped into the sample tank, DNA hybridization is performed, and the labeling substance that has not contributed to the reaction is removed with a buffer solution (PBS (PBS)). Wash with a mixture of phosphoric acid buffer), EDTA (ethylene sodium acetate sodium nitrate), and NaCl (pH 7.4).

 The sample obtained in this way is set on the sample stage 1 shown in FIGS. 11A and 11B. Then, the sample tank 201 is irradiated with excitation light from each LED light source unit 41, 42, 43. At this time, the light from the LED light source having three different wavelengths as the peak emission wavelengths is simultaneously irradiated onto the sample surface.

Then, fluorescence is emitted from the fluorescent dye labeled on the sample that has reacted with the DNA probe by the DNA hybridization, and the light passes through the objective lens 21 shown in Fig. 12 and is diced. Reutz Miller 4 5 is reached. In this case, the fluorescence from the fluorescent dye FITC is reflected by the dichroic mirror 45, passes through the lens 47, enters the CCD camera 48, and is guided to the computer 10. It is obtained as a green fluorescent image by FITC. The fluorescence of the fluorescent dye Cy 3 passes through the dichroic mirror 45, is reflected by the dichroic mirror 46, passes through the lens 49, and passes through the lens 49. Enters 0, is led to computer 10 and emits orange fluorescence due to .Cy3 Obtained as an image. Further, the fluorescence of the fluorescent dye Cy 5 passes through the dichroic mirror 45 and the dichroic mirror 46, respectively, passes through the lens 51, and passes through the CCD 51. After entering the camera 52, it is guided to the computer 10 and a red fluorescent image by Cy5 is obtained. The computer 10 synthesizes the fluorescent images from the three types of fluorescent dyes by the CCD cameras 48, 50, and 52 and outputs them to the monitor 13 or outputs them from the three types of fluorescent dyes. Display the fluorescent images separately on monitor 13.

 FIG. 13 shows a block diagram of the drive circuits of the first to third LED light source units 41, 42, and 43.

 In this case, as the driving circuits of the first to third LED light source units 41, 42, and 43, an LED light source unit driving circuit 54 and an LED light source unit driving circuit 55 are respectively provided. An LED light source unit driving circuit 56 is provided. The LED light source unit drive circuit 54 is connected to the first LED light source unit 41 and the LED light source unit drive circuit 55 is connected to the second LED light source unit 42. An LED light source unit drive circuit 56 is connected to the third LED light source unit 43. A common power supply 53 is connected to the LED light source unit driving circuit 54, the LED light source unit driving circuit 55, and the LED light source unit driving circuit 56, and as described in FIG. Combiner 10 is connected.

The LED light source unit drive circuit 54, the LED light source unit drive circuit 55, and the LED light source unit drive circuit 56 receive power from a common power supply device 53. Also, LED light source- The LED drive circuit 54, the LED light source unit drive circuit 55, and the LED light source unit drive circuit 56 are controlled based on a command from the computer 10, and are controlled by the first LED light source unit. A driving current is supplied to each of the LED light sources of the light source 41, the second LED light source unit 42, and the third LED light source unit 43 to generate excitation light of three kinds of fluorescent dyes.

 In this case, as shown in FIG. 12, the three types of fluorescence signals obtained by these excitation lights are respectively transmitted by the CCD cameras 48 and 46 by the dichroic mirrors 45 and 46, respectively. , 50 and 52 separately, and the image output signals from these CCD cameras 48, 50 and 52 are guided to the computer 10, where image analysis is performed, and three types of images are obtained. A composite image using the fluorescent dye is generated and output.

 FIG. 14 shows a specific circuit configuration of the power supply unit 53 and the LED light source unit drive circuits 54, 55, and 56.

In this case, the power supply device 53 guides the output from the AC power supply 57 of 100 V to the transformer circuit 58 composed of a transformer to adjust the voltage. The output from the transformer circuit 58 is sent to a bridge rectifier circuit 59 composed of four diodes to perform full-wave rectification, and then smoothed by a smoothing capacitor 60. The smoothed output is led to a constant voltage circuit 6 4 composed of two power transistors 6 16 2 connected in Darlington and an OP amplifier 63, and a constant voltage is output. Is output. In this case, the terminal voltage of the Zener diode 65 is used as a reference voltage, and a variable reference voltage is generated by a variable reference voltage generation circuit 67 composed of circuit elements such as a P-amplifier 66 and a resistor. are doing. And the OP W

By changing the value of the variable reference resistor 68 connected to the amplifier 63, the voltage output of the constant voltage circuit 64 is adjusted in relation to the variable reference voltage of the variable reference voltage generation circuit 67. Then, a drive current is supplied to the LED light source unit drive circuit 54, the LED light source unit drive circuit 55, and the LED light source unit drive circuit 56. In other words, by changing the value of the variable reference resistor 68, the driving current to the LED light source unit driving circuit 54, the LED light source unit driving circuit 55, and the LED light source unit driving circuit 56 is increased. Can be adjusted at the same time. In this case, a FET (field effect transistor) 70 generates an abrupt current in the event of a short-circuit when the current is short-circuited, and flows through the OP amplifier 63. It plays a role in preventing accidental damage. The power can be turned on and off by the power switch 69. The power supply stitch 69 is connected to the computer 10, and the power input is also adjusted by the computer 10. The power supply switch 69 may be manually switched.

To such a power supply 53, LED light source unit driving circuits 54, 55, 56 are connected in parallel. The LED light source unit drive circuit 54 serves as a drive circuit for the four first LED light source units 41 described in FIGS. 11A and 11B, and includes a variable resistor 71 and a variable resistor 71. scan I Tutsi 7 2 _t Similarly the series circuit are provided separately to each, LED light source unit drive circuit 5 5 also as the second LED light source Interview two Tsu DOO 4 2 of the driving circuit of the four partial A series circuit of a variable resistor 71 and a switch 72 is separately prepared, and the LED light source unit driving circuit 56 is also equipped with a third LED light source unit for four. As a drive circuit for the unit 43, a series circuit of a variable resistor 71 and a switch 72 is separately provided.

 In these LED light source unit driving circuits 54, 55, and 56, the first to third LED light source units 4 are controlled by adjusting the resistance values of the respective variable resistors 71, 73, and 75. The drive current supplied to each of the LED light sources 1, 42, and 43 can be adjusted, and the output light intensity can be controlled. The adjustment of the output light intensity may be performed manually, or may be automatically performed automatically by a computer. This makes it possible to individually control the drive current of each of the first to third LED light source units 41, 42, and 43.

 The LED light source unit driving circuits 54, 55, and 56 control the on / off of the switches 72, 74, and 76 to control the first to third LED light source units 4. The LED light sources 1, 42, and 43 can all be turned on and off independently. These on / off controls may be performed automatically by computer control or manually.

When adjusting the resistance values of the variable resistors 71, 73, and 75 by computer control, the first to third LED light source units 4 as described with reference to FIG. The light intensity from each LED light source of 1, 42, 43 is detected, the detection signal is led to a computer, and the light intensity is analyzed by a computer, and the variable resistors 71, 73, 7 Adjust the resistance of 5. Alternatively, the resistance values of the variable resistors 71, 73, and 75 may be manually adjusted based on the light intensities from the individual LED light sources obtained by this detection. This Thereby, it is possible to adjust so that the light from the LED light source is almost uniformly irradiated to a part of the reaction tank of the DNA microarray. Further, it is possible to partially adjust the light intensity, for example, by concentrating the excitation light at a desired position in the sample tank 201. Further, the variable resistances 71, 73, and 75 can control the magnitude of the drive current flowing through all LED light sources, so that the intensity of the excitation light can be increased or reduced all at once. You can do it. As a result, it is possible to irradiate each sample with excitation light of an appropriate intensity, and it is possible to efficiently detect fluorescence.

 Therefore, according to this method, three types of fluorescent dyes can be formed by using the first to third LED light source units 41, 42, and 43 having three types of LED light sources having different peak emission wavelengths. Since simultaneous excitation is possible, fluorescence images from these three types of fluorescent dyes can be acquired, and these can be synthesized or displayed individually.

 In addition, the individual LED light sources constituting the first to third LED light source units 41, 42, and 43 can easily be turned on and off according to the fluorescent dye to be excited. In addition, the drive current can be individually adjusted by the variable operation of the variable resistance, so that each fluorescent dye can be irradiated with excitation light of the correct intensity, and the efficiency can be improved. Fluorescence can be detected well.

In the above-described third embodiment, three types of fluorescent dyes are used. However, the present invention is not limited to this, and two or more types of fluorescent dyes may be obtained in the same manner. In Wear. That is, it is only necessary to add dichroic mirrors, lenses, and CCD force lenses for each type of fluorescent dye. Also, the fluorescent dye is not limited to FITC, Cy3, and Cy5 shown in the examples, but may be a load-min green (Rhodamine G), a Texas-red (Texas). Red), RITC (Rhodamine B-isothiocya), etc. may be used.The number of LED light source units is not particularly limited, and a plurality of LED light source units are individually controlled by a plurality of drive circuits. I can do it. Further, the present invention can be applied even if the output wavelengths of a plurality of LED light source units are the same or different. It is also possible to irradiate the sample with one type of light from LED light sources having different peak emission wavelengths, and to repeat the light several times at different wavelengths. For example, it is possible to use a turret in which filters with different transmission wavelengths are set and automatically switch by computer in conjunction with LED light sources of different wavelengths. . .

 (Fourth embodiment)

 In the fourth embodiment, a reflecting member is arranged on the optical path of light emitted from the LED light source unit, and the light in the optical path is reflected, so that the optical axis angle of the excitation light can be increased and the light of the objective lens can be increased. This is an embodiment that approaches an axis angle.

 FIG. 15 is a diagram showing a schematic configuration of the fourth embodiment, and the same parts as those in FIG. 1 are denoted by the same reference numerals.

In this case, around the objective lens 5, a plurality of LED light source units 81 as semiconductor light source means (four LED light units in the illustrated example) are used. D light source units 81, one of which is not shown), are arranged radially around the optical axis of the objective lens 5.

 The LED light source unit 81 has a cylindrical LED light source holder 8101. The LED light source holder 8101 has an LED light source 8102 disposed therein. In addition, on the optical path of light emitted from the LED light source 8102 inside the LED light source holder 8101, a bandpass filter 8103, a diffusion plate 8104, a condenser lens 8 1 0 5 is arranged. Here, the non-noise filter 8103 mainly passes near the wavelength range of light emitted from the LED light source 8102.

 On the optical path of the light emitted from the LED light source unit 81, reflectors 82 are provided as reflecting members. The reflecting plate 82 reflects light from the LED light source unit 81 and irradiates the sample 202 in the sample tank 201 on the sample stage 1 as excitation light. At the same time, the reflector 82 reduces the angle す formed by the optical axis of the excitation light with respect to the optical axis of the objective lens 5 so that the optical axis angle of the excitation light approaches the optical axis angle of the objective lens. I'm doing it.

 Here, the respective mounting positions of the reflectors 8 2 reflect the light from the LED light source unit 81 and irradiate the sample 20 ′ 2 surface with uniform brightness and irradiate the light. In order to prevent excitation unevenness from occurring on the sample 202 surface due to the incident angle of light, shadows and excitation unevenness also occur on the sample 202 having a three-dimensional part on part or the entire surface. It is adjusted so that it does not occur.

 Others are the same as in Fig. 1.

Therefore, even in this case, the same effect as in the first embodiment is obtained. You can expect results. In addition, a reflector 82 is arranged on the optical path of the light emitted from the LED light source unit 81 to reflect the light in the optical path, so that the optical axis angle of the excitation light can be adjusted by the objective lens 5. It can approach the optical axis angle. This means that the excitation light can be radiated from a position close to the top of the sample surface 202 (the limit that does not enter the observation field of view of the objective lens 5), so that the LED light source unit The difference between the shortest distance a and the longest distance b of the optical path from 81 to the sample 202 surface through the reflector 82 can be reduced. As a result, since the intensity of the excitation light is inversely proportional to the distance of the optical path, the difference in the intensity of the excitation light on the surface of the sample 202 can be reduced. The two surfaces can be excited efficiently and uniformly.

 It should be noted that also in the fourth embodiment, a control method for illuminating a sample uniformly and uniformly with light as described in the modification of the second embodiment and the third embodiment. Can be adopted.

 (First modification)

 In the fourth embodiment, the reflection plate 82 is arranged as a reflection member on the optical path of the light emitted from the LED light source unit 81 to reflect the light in the optical path. If such a method is used, the degree of freedom in arranging the LED light source units 81 increases, so that a large number of LED light source units 81 are arranged around the objective lens 5. It can be.

FIG. 16 is a diagram showing a schematic configuration of an optical inspection apparatus according to a first modification of the fourth embodiment. The same parts as those in FIG. The label is attached.

 Around the objective lens 5, a plurality of LED light source units 81 as semiconductor light source means are arranged in multiple layers (two layers in the illustrated example). In addition, on the optical path of the light emitted from these LED light source units 81, reflecting plates 82, 82 '(corresponding to the LED light source units 81 of each layer) as reflecting members. (Corresponding to 2 layers in the example shown). The reflectors 82 and 82 'reflect light from the LED light source unit 81 of each layer and serve as excitation light for the sample 202 in the sample tank 201 on the sample stage 1. And irradiate.

 In this case, the light from the LED light source unit 81 is reflected to irradiate the sample 202 with uniform brightness even at the respective mounting positions of the reflectors 82 and 82 '. In addition, in order to prevent excitation unevenness from occurring on the sample 202 surface due to the incident angle of the irradiated light, the sample 202 having a three-dimensional portion on part or the entire surface is also shaded. Fig. 15 is the same as Fig. 15 for other adjustments so that no excitation unevenness occurs.

According to the first modified example of the fourth embodiment, the light from the light source unit 81 of each layer is reflected by using the reflectors 82, 82 '. In addition, it is possible to determine the degree of freedom of the position where the LED light source unit 81 of each layer is arranged. Since the degree of freedom of the position where these LED light source units 81 are arranged is increased, more LED light source units 8 than in the fourth embodiment are provided. It becomes possible to use 1. This makes it easier to obtain the amount of light required to irradiate the sample 202 surface. In addition, it is possible to illuminate the sample 202 surface without unevenness in light quantity.

 In addition, since the number of LED light source units 81 can be greatly increased, these LED light source units 81 are divided into a plurality of sets, and the emission wavelength spectrum is increased for each set. By using LED light sources with different torque characteristics, it is possible to excite a plurality of types of fluorescent dyes and detect fluorescent signals in the same manner as described in the third embodiment. And

 (Second modification)

 In the fourth embodiment, the reflection plate 82 is disposed as a reflection member on the optical path of the light emitted from the LED light source unit 81 to reflect the light in the optical path. A die crack mirror can be used as a member.

 FIG. 17 is a diagram showing a schematic configuration of the second modification, and the same parts as those in FIG. 15 are denoted by the same reference numerals.

 The LED light source unit 8 1 has an LED light source 8 1 0 2 disposed inside a cylindrical LED light source holder 8 1 0 1 and an LED light source 8 1 0 inside the LED light source holder 8 1 0 1. A diffusing plate 8104 and a condensing lens 8105 are arranged on the optical path of the light emitted from 2, and the above-described bandpass finoleta 8103 is omitted.

On the optical path of the light emitted from the LED light source unit 81, a die-cloth mirror 83 is arranged as a reflecting member. The dichroic mirror 8 3 mainly reflects near the wavelength range of the light emitted from the LED light source 8 102 of the LED light source unit 81. It has such characteristics. The die-cloth mirror 83 reflects the light from the LED light source unit 81 and applies excitation light to the sample 202 in the sample tank 201 on the sample stage 1. And irradiate. Also in this case, the angle between the optical axis of the excitation light and the optical axis of the objective lens 5 is reduced so that the optical axis angle of the excitation light approaches the optical axis angle of the objective lens.

 Others are the same as in Fig. 15.

 In this way, the same effect as that of the fourth embodiment can be expected even if the die mirror mirror 83 is used instead of the reflection plate as the reflection member. You. Furthermore, the die-cloth mirror 83 has a characteristic of mainly reflecting near the wavelength range of light emitted from the LED light source 8102 of the LED light source unit 81. By using the LED, it is possible to omit the bandpass finolators built in the LED light source unit 81. As a result, the cost of the LED light source unit 81 can be reduced and the size can be reduced. (Fifth embodiment)

 In the fifth embodiment, the light guide member is arranged on the optical path of the light emitted from the LED light source unit, and the optical path is bent, so that the optical axis angle of the excitation light is increased and the optical axis angle of the objective lens is increased. This is an embodiment in which the distance is approached.

 FIG. 18 shows a schematic configuration of the fifth embodiment, and the same parts as those in FIGS. 1 and 15 are denoted by the same reference numerals.

In this case, around the objective lens 5, a plurality of LEDs (four in the illustrated example, one of which is not shown) as semiconductor light source means are provided. The light source unit 81 is arranged.

 The LED light source unit 81 has a cylindrical LED light source holder 8101. The LED light source holder 8101 has an LED light source 8102 disposed therein. In addition, on the optical path of the light emitted from the LED light source 8102 inside the LED light source HONOLEDA 8101, a bandpass filter 8103, a diffusion plate 8104, 810 5 are arranged. Here, the non-pass filter 8103 mainly transmits light near the wavelength range of the light emitted from the LED light source 8102.

 On the optical path of the light emitted from the LED light source unit 81, a light incident end 84a of an optical fiber 84 is arranged separately as a light guide member. The optical fiber 84 can bend the optical path of the light from the LED light source unit 81 freely. As a result, the light from the light emitting end 84b is irradiated as the excitation light onto the sample 202 in the sample tank 201 on the sample stage 1, and at the same time, the light from the objective lens 5 The angle formed by the optical axis of the excitation light with respect to the optical axis can be reduced so that the optical axis angle of the excitation light approaches the optical axis angle of the objective lens.

 In this case, the position of the light emitting end 84b of these optical fibers 84 is caused by the fact that the emitted light is applied to the sample 202 surface with uniform brightness and the incident angle of the applied light. Adjustment so that excitation unevenness does not occur on the sample 202 surface, and also so that shadows and excitation unevenness do not occur on the sample 202 having a three-dimensional part on all or part of the surface. Have been.

Others are the same as in FIGS. 1 and 15. Therefore, in the fifth embodiment, the same effects as in the first embodiment can be expected. Further, since the optical path is bent by the optical fiber 84 arranged on the optical path of the light emitted from the LED light source unit 81, the optical axis angle of the excitation light is adjusted by the objective lens. It can approach the optical axis angle. As a result, similarly to the fourth embodiment, the difference in the intensity of the excitation light on the sample 202 surface can be reduced, and the sample 202 surface can be efficiently used. It can be excited uniformly.

 In the present embodiment, an example in which the optical guide member 84 is used as the light guide member has been described. However, for example, a material obtained by molding glass rod-shaped acrylic material is applied. You can also do it. It should be noted that also in the fifth embodiment, a control method for illuminating a sample uniformly and uniformly with light as described in the modification of the second embodiment and the third embodiment. Can be applied.

 (Modification)

 In the fifth embodiment, the optical fiber 84 is arranged as a light guide member on the optical path of the light emitted from the LED light source unit 81, and the optical path is bent. If such a method is used, the degree of freedom in arranging the LED light source units 81 increases, so that it is possible to arrange a large number of LED light source units 81 around the objective lens 5. it can.

FIG. 19 is a diagram showing a schematic configuration of an optical inspection device according to a modification of the fifth embodiment, and the same parts as those in FIG. 18 are denoted by the same reference numerals. In FIG. 19, around the objective lens 5, a plurality (two in the illustrated example) of a plurality of LED light source units 81 as semiconductor light source means are arranged. In addition, on the optical path of the light emitted from the LED light source unit 81, a light incident end 84a of an optical canopy refino 84 as a light guide member is separately arranged. The optical fiber 84 allows the light path of the light from the LED light source unit 81 to be freely bent, and the light from the light emitting end 84b is transferred to the sample on the sample stage 1. The sample 202 in the tank 201 is irradiated as excitation light.

 Also in the case of this modified example, the position of the light emitting end 84b of the optical fiber 84 is such that the emitted light is applied to the surface of the sample 202 with uniform brightness, and the light to be applied is In order to prevent excitation unevenness from occurring on the sample 202 surface due to the incident angle, no shadow or excitation unevenness occurs on the sample 202 having a three-dimensional part on all or part of the surface. It has been adjusted accordingly.

 Others are the same as in Fig. 18.

In this modification, the optical path of the light from the light source unit 81 is bent by using the optical fin 84, so that the position of each LED light source unit 81 is automatically determined. The degree of freedom can be increased. By increasing the degree of freedom in the location of the LED light source units 81, it is possible to use more LED light source units 81 than in the fifth embodiment. And are possible. This makes it possible to easily obtain the amount of light required to irradiate the sample 202 surface, and to illuminate the sample 202 surface without unevenness in the light amount. In addition, since the number of LED light source units 81 can be greatly increased, the LED light source units 81 are divided into a plurality of sets, and the spectral characteristics of the emission wavelength are set for each set. By using different LED light sources, it is also possible to excite a plurality of types of fluorescent dyes and detect fluorescent signals as in the third embodiment.

 In this modification, an example in which the optical fiber 84 is used as the light guide member has been described. However, for example, a material obtained by molding a glass rod material may be used. it can.

 The present invention is not limited to the above-described embodiment, and can be variously modified in the implementation stage without departing from the spirit of the invention. For example, the sample applied in each of the above-described embodiments is not limited to an example of a DNA microarray, but can be applied to all reaction vessels called so-called DNA microarrays. Further, the present invention is widely applicable not only to DNA but also to detection and measurement dealing with various other biological materials.

 Furthermore, the above-described embodiment includes various stages of the invention, and various inventions can be extracted by appropriately combining a plurality of disclosed constituent elements. For example, even if some components are deleted from all the components shown in the embodiments, the problem described in the section of the problem to be solved by the invention can be solved, and the problem described in the section of the effect of the invention can be solved. If the effect obtained is obtained, a configuration from which this component is deleted can be extracted as an invention.

According to the embodiment of the present invention, light is applied to the labeled substance of the sample. Since the optical path for irradiation and the optical path of the fluorescence detecting means having the focusing lens are separated, optical elements such as dichroic mirrors that attenuate light can be omitted, and the detected light intensity Loss can be minimized.

 Further, according to the embodiment of the present invention, a plurality of semiconductor light source means are arranged along the periphery of the converging lens and light is irradiated simultaneously from these, so that the labeling substance is irradiated with light. It is possible to efficiently obtain light of sufficient intensity to carry out.

 According to the embodiment of the present invention, the direction of the light emitted from the semiconductor light source means and the direction of inclination of the condenser lens with respect to the optical axis can be adjusted, respectively, so that the desired specific Light can be condensed at the location, and efficient irradiation of the labeling substance with light can be performed.

 Furthermore, according to the embodiment of the present invention, since the drive current to the semiconductor light source means can be individually controlled based on the light intensity obtained near the sample, the light from the semiconductor light source means can be controlled by the same light. Brightness can be adjusted, and the sample can be illuminated with uniform brightness. '

 As described above, according to the embodiment of the present invention, it is possible to provide a photodetector capable of efficiently irradiating sufficient light and performing highly reliable photodetection. .

Industrial applicability

 The present invention relates to a photodetection device that can efficiently irradiate sufficient light and can perform highly reliable photodetection.

Claims

The scope of the claims

 1. The photodetector is

 A semiconductor light source means for emitting light for irradiating the sample with a labeled substance with light,

 A condensing lens for condensing the light emitted by irradiating the labeling substance with light by the irradiation light, and a light emitted by irradiating the labeling substance with light by the irradiation light; And a photodetector having a filter that transmits the light, and a photodetector that detects light that has passed through the focusing lens.

 The optical path of the irradiation light is an optical path different from the optical path of the light detecting means.

 2. The photodetector according to claim 1, wherein the semiconductor light source means includes an LED light source that emits irradiation light and a light source from the LED light source that is arranged on an optical path of light emitted from the LED light source. It has at least one of a filter that selectively transmits wavelengths, a diffuser, and a lens.

 3. The photodetector according to claim 1 or 2, wherein the semiconductor light source is disposed around the focusing lens, and the semiconductor light source is held around the focusing lens. Semiconductor light source means holding means for holding the semiconductor light source means at a predetermined angle with respect to the optical axis of the focusing lens so that light emitted from the means is irradiated on the sample;

The semiconductor light source holding means is provided at a holding portion of the semiconductor light source means, and at least one of a direction of light emitted from the semiconductor light source means and an inclination direction of the condenser lens with respect to an optical axis is provided. And a position adjusting means for adjusting the direction.

 4. In the photodetector according to claim 1 or 2, the semiconductor light source means has a plurality of semiconductor light source units arranged along the periphery of the condenser lens.

 5. The photodetector according to claim 4, wherein the plurality of semiconductor light source units have spectral characteristics of substantially the same emission wavelength.

 6. The photodetector according to claim 4, wherein the plurality of semiconductor light source units have spectral characteristics of different emission wavelengths.

 7. The light detection device according to any one of claims 4 to 6, wherein

 The plurality of semiconductor light source units are arranged around the condenser lens, and the plurality of semiconductor light source units are held around the condenser lens, and light emitted from the plurality of semiconductor light source units is emitted from the plurality of semiconductor light source units. Semiconductor light source means holding means for holding the plurality of semiconductor light source units at a predetermined angle with respect to the optical axis of the focusing lens so as to irradiate the sample;

 The semiconductor light source holding means is provided at a holding portion of the plurality of semiconductor light source units, and has a small direction of light emitted from the plurality of semiconductor light source units and a small inclination direction with respect to an optical axis of the focusing lens. In addition, it further has a position adjusting means for adjusting one of them.

8. The photodetector according to any one of claims 4 to 7, wherein the semiconductor light source means comprises: The central axis of the light emitted from the step is arranged so as to pass through the periphery of the sample and intersect at one point of a vertical position above or below the substantially central axis of the sample.

 9. The photodetector according to any one of claims 4 to 7, wherein the semiconductor light source means is configured such that a center axis of light emitted from the semiconductor light source means is shifted from a center axis of the sample. At a point above the space position vertically above or below the sample.

 10. The photodetector according to claim 1, wherein the photodetector is disposed on an optical path of light emitted from the plurality of semiconductor light source units, reflects light in the optical path, and emits light to the sample. And a reflecting member for irradiating light.

 11. The photodetector according to claim 1 or 2, wherein the photodetector has a characteristic of reflecting near a wavelength range of light emitted from the semiconductor light source means, and detects light emitted from the semiconductor light source means. The apparatus further includes a dichroic mirror disposed on the optical path for reflecting light in the optical path and irradiating the sample with light.

 12. The photodetector according to claim 1 or 2, wherein the light source is arranged in an optical path of light emitted from the semiconductor light source means, and the optical path is bent to irradiate the sample with light in the optical path. The light guide member is further provided.

 13. The photodetector according to any one of claims 10 to 12, wherein the semiconductor light source means has a different spectral characteristic of an emission wavelength.

14. The light according to any one of claims 1 to 13. In the detection device,

 Light intensity detection means for detecting the intensity of light applied to the sample in the vicinity of the sample;

 A driving means for supplying a driving current for driving the semiconductor light source means,

 The driving unit variably controls a driving current to the semiconductor light source unit based on the intensity of light detected by the light intensity detecting unit.

15. The photodetector according to any one of claims 4 to 9, wherein:

 Light intensity detecting means for detecting the intensity of light applied to the sample in the vicinity of the sample;

 A driving means for supplying a driving current for driving the semiconductor light source means,

 The driving means is capable of individually controlling a driving current to the semiconductor light source means based on the intensity of light detected by the light intensity detecting means.

 1 6. The photodetector is

 A semiconductor light source means for irradiating light to the labeled substance of the sample, the semiconductor light source means includes: a semiconductor light emitting element; and an optical element,

A condenser lens for condensing light emitted by irradiating the labeling substance with the light by the irradiation light; and light emitted by irradiating the labeling substance with light by the irradiation light. And a photodetector having a photodetector that detects light that has passed through the condenser lens.

17. The photodetector according to claim 16, wherein the optical element includes at least one of a band filter, a diffuser, and a condenser lens.