Luminescence detection device

Abstract

The luminescence detection device of the present invention uses each focusing lens of the focusing lens array to individually focus the light from each light-emitting point of the luminescence point array to make it into a light beam and make each light beam enter the sensor in parallel for detection. The luminescence detection device is made compact while achieving high sensitivity and low crosstalk by ensuring that the diameter of each light-emitting point, the focal length and interval of each focusing lens, and the optical path length of each focusing lens and the sensor satisfy a predetermined relationship.

Description

[0001] This application is a divisional application; the application number of the parent application is “2016800812872”, and the name of the invention is “Luminescence Detection Device”. Technical Field

[0002] The present invention relates to a detection device that irradiates a plurality of channels provided inside a plurality of capillaries or a microchip with light such as a laser beam or a lamp, and detects with high sensitivity fluorescence, phosphorescence, scattered light, or transmitted light emitted by a substance present inside the capillaries or the channels. Background Art

[0003] The following capillary array DNA sequencer is widely used: by parallel processing of electrophoresis analysis of multiple capillaries (glass capillaries with an outer diameter of 100 to 400 μm and an inner diameter of 25 to 100 μm) filled with separation media, the base sequence of different DNA samples is read in each capillary at the same time. The mechanism is described below. Since the capillaries sold on the market are flexible, a polyimide covering film is formed on the outer surface. The portion of the electrophoresis path of each capillary with a constant length, such as the position of the capillary 30 cm away from the sample injection end, is aligned and arranged on the same plane in a state after the covering film is removed, and a laser beam is irradiated from the side along the above-mentioned capillary arrangement plane, thereby irradiating multiple capillaries at the same time. Hereinafter, in this specification, the above-mentioned capillary arrangement plane is sometimes simply referred to as the arrangement plane. When the laser beam passes through, the fluorescent-labeled DNA electrophoresed inside each of the above-mentioned capillaries is excited by the laser irradiation and emits fluorescence. Here, DNA is colored according to the types of terminal bases of A, C, G, and T and is divided into four colors of fluorescent bodies. As a result, the laser irradiation position of each capillary becomes a luminous point, and a plurality of luminous points are arranged in a straight line at intervals of p. Hereinafter, it is referred to as a luminous point array. When the number of luminous points (the number of capillaries) is set to M, the full width AW of the luminous point array is AW=p×(M-1). For example, when p=0.36mm and M=24, AW=8.28mm. The luminescence detection device detects each light emission from the luminous point array while splitting the light. The structure of the device is shown in Patent Document 1.

[0004] First, each light emission is made into a parallel beam by a common condenser lens. Hereinafter, the expression "shared" means that a plurality of (M) light-emitting points use one optical element (M:1 correspondence). On the contrary, the expression "individual" means that one light-emitting point uses one optical element (1:1 correspondence). Here, when the focal length of the common condenser lens is set to f1 and the effective diameter is set to D1, it is necessary to set AW<f, AW<D1. For example, the condenser lens is a camera lens with f1=50mm and D1=36mm. Next, the parallel beam is passed through a long pass filter to cut the wavelength of the laser beam, and is made to pass through a common transmission diffraction grating to disperse the wavelength in the long axis direction of each capillary, that is, the direction orthogonal to the arrangement direction of the light-emitting point array and the optical axis of the common condenser lens. Here, when the effective diameter of the common transmission diffraction grating is set to DG, in order not to reduce the detection efficiency, D1≤DG is required. For example, DG=50mm. Next, each parallel light beam is imaged on a two-dimensional sensor by a common imaging lens. Here, when the focal length of the common imaging lens is set to f2 and the effective diameter is set to D2, in order not to reduce the detection efficiency, D1≤D2 is required. For example, the imaging lens is a camera lens with f2=50mm and D2=36mm. In summary, the wavelength dispersion spectrum image of each light emitted from the light-emitting point array can be obtained at the same time. Finally, the time variation of the fluorescence intensity of the four colors is obtained by analyzing the time variation of each wavelength dispersion spectrum, thereby determining the order of the base types, that is, the base arrangement.

[0005] Non-patent document 1 shows another scheme for simultaneously detecting four colors of fluorescence. First, a focusing lens (here, an objective lens) is used to make the light emitted from a light-emitting area into a parallel beam. Here, when the full width of the light-emitting area is set to AW, the focal length of the objective lens is set to f, and the effective diameter is set to D, AW<f, AW<D. The objective lens used is Olympus's UPLSAPO 60×W, and AW=0.44mm, f1=3mm, D1=20mm. Next, a group of three color separation mirrors split the parallel beam into four parallel beams of four colors. Next, a group of four imaging lenses are used to image each parallel beam on four two-dimensional sensors. Here, when the effective diameter of each imaging lens is set to D2, in order not to reduce the detection efficiency, D1≤D2 is required. In summary, the four-color four-split image of the light-emitting area can be obtained at the same time.

[0006] On the other hand, Patent Document 2 shows another scheme for simultaneously detecting the light emitted from the light-emitting point array. First, the light emitted from the light-emitting point array is individually converted into parallel light beams by a focusing lens array. Here, when the interval between the light-emitting points is set to p and the number of light-emitting points is set to M, the full width of the light-emitting point array is AW=p×(M-1), and when the effective diameter of each focusing lens is set to D, D<AW, which is a different structure from the above-mentioned Patent Document 1 and Non-Patent Document 1. In addition, by setting D<p, each focusing lens can become an individual focusing lens array arranged in a straight line. Next, each parallel light beam is incident on the individual sensor of each individual sensor array. In summary, the light emission intensity from the light-emitting point array can be obtained at the same time.

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: Japanese Patent Application Publication No. 2007-171214

[0010] Patent Document 2: Japanese Patent Application Publication No. 2011-59095

[0011] Non-patent literature

[0012] Non-patent literature 1: Rev Sci Instrum., 2011 Feb; 82(2): 023701. Summary of the invention

[0013] Problems to be solved by the invention

[0014] The luminescence detection device of Patent Document 1 has high condensing efficiency (condensing efficiency of a common condensing lens) and detection efficiency (total utilization efficiency of luminescence that contributes to fluorescence detection by the sensor based on the condensing efficiency, transmittance of a long-pass filter, diffraction efficiency of a diffraction grating, etc.), and high spectral accuracy of the diffraction grating. The condensing efficiency can be represented by the F value of a common condensing lens, which is related to 1 / F. 2Proportional. For example, when a camera lens with f1=50mm and D1=36mm is used, F=f1 / D1=1.4, and high-sensitivity luminescence detection can be performed. In addition, since the luminescence from each luminous point is imaged by a common imaging lens and then detected by a two-dimensional sensor, the luminescence will not be mixed with each other, that is, each luminescence can be detected independently with low crosstalk. The above high sensitivity and low crosstalk performance are particularly important in the field of high-precision detection of trace measurement objects, or simultaneous and independent detection of multiple different measurement objects. However, the luminescence detection device includes two common lenses, and there is a relationship of AW<f, AW<D1≤D2. Therefore, if AW is set to be constant, there is a problem that the overall size of the device is very large and the manufacturing cost of the device is high. For example, when a camera lens with f1=f2=50mm and D1=D2=36mm is used, the overall size of the luminescence detection device is smaller than the volume of a cylinder with a diameter of 100mm and a height of 200mm (1.6×10 6 mm 3 ) is large. In this specification, the overall size of the light detection device is expressed by the volume occupied by the optical system from the light-emitting point to the sensor, and the volume occupied by the sensor itself is not included. In addition, since it is impossible to set AW<<f1 and AW<<D1 (a huge camera lens is required to achieve the above situation), the detection efficiency of the light-emitting point far from the optical axis (the light-emitting point located near the end of the light-emitting point array) is lower than the detection efficiency of the light-emitting point near the optical axis (the light-emitting point located near the center of the light-emitting point array), resulting in a problem of sensitivity deviation in each light-emitting point.

[0015] However, these problems have not been solved so far, that is, the device for simultaneously identifying and detecting the luminescence of four colors from the luminescent point array has not been made small and the cost has not been reduced, and the sensitivity deviation of each luminescence has not been reduced. If the luminescence detection device can be made small, the capillary array DNA sequencer can be set in a smaller area or can be carried, or the ease of use can be improved. In addition, the manufacturing cost can be reduced by reducing the number of components of the detection device or reducing the size of each component. In addition, by reducing the sensitivity deviation of each luminescent point, quantitative comparison of the analysis sample can be performed in each capillary, so that the total sensitivity and dynamic range can be improved in the luminescence detection of the luminescent point array. As the above result, the capillary array DNA sequencer is further popularized, so that it can further contribute to the world.

[0016] Using the luminescence detection device shown in non-patent document 1, it is possible to simultaneously perform fluorescence detection on four colors of luminescence from the same luminescent point array. However, in the objective lens used in non-patent document 1, since AW = 0.44 mm, for example, only a very small part of the full width of the luminescent point array of 8.28 mm can be detected. Therefore, a common focusing lens and four common imaging lenses that are the same as those of the capillary array DNA sequencer are used to replace the objective lens and the four imaging lenses. At this time, when the effective diameter of the three color separation mirrors is set to DM, since they are configured at a 45° tilt relative to the parallel light beam, in order not to reduce the detection efficiency, √2×D1≤DM is required. For example, DM = 71 mm. Therefore, even if the four two-dimensional sensors are not included, the overall size of the luminescence detection device is larger than that of patent document 1, and the manufacturing cost is correspondingly higher. In addition, the space occupied by the four two-dimensional sensors is large, and their cost is also very high. As a result, there is still the issue of sensitivity deviation of each luminescent point.

[0017] On the other hand, when using the light-emitting detection device shown in Patent Document 2, since D<AW, it is possible to reduce the size of the device. However, as described in the document, when individual focusing lens arrays are used to focus and detect the light emitted from the light-emitting point array, it is difficult to obtain both high sensitivity and low crosstalk performance, which are in a trade-off relationship. In the document, there is no record of such a problem and the discussion and consideration of the solution to the problem. Next, the above-mentioned problem is described in detail.

[0018] As described above, when the focal length of each condenser lens is f and the effective diameter is D, the condensing efficiency can be expressed by F=f / D (the condensing efficiency is equal to 1 / F). 2 Here, if D is set to be constant, the smaller f is, that is, the closer the light-emitting point is to the focusing lens, the higher the focusing efficiency will be. This is correct when the size of the light-emitting point is infinitely small, but not necessarily correct when it is finite. Here, when the diameter of the light-emitting point is set to d, infinitesimal refers to the case where d<<f or d≈0, and finite refers to the case where d>0. In fact, it can also be set to d≥0.01mm. Below, in the present invention, the case where the size of the light-emitting point is finite is considered. If the center of the light-emitting point is placed at the focal position of the lens, the light emitted from the center of the light-emitting point becomes a parallel light beam due to the lens, and the light beam travels along the optical axis of the lens in a direction parallel to the optical axis. On the other hand, the light emitted from the end of the light-emitting point becomes a parallel light beam due to the lens, and the light beam travels along an angle θ=tan -1(d / 2 / f) direction. That is, these light beams separate as they move away from the lens. Therefore, when a sensor (or a part of the area of ​​the allocated area sensor) of the size of the parallel light beam emitted from the center of the light-emitting point is arranged at a certain distance from the lens on the optical axis of the lens to detect light emission, all light emitted from the center of the light-emitting point can be detected, and only a part of the light emitted from the end of the light-emitting point can be detected. Since the larger f is, the smaller the above angle θ is, the greater the proportion of light emission that can be detected.

[0019] As described above, in order to improve the light-gathering efficiency and obtain high sensitivity, it is known that there is a trade-off relationship between preferably reducing f and preferably increasing f. However, including Patent Document 2, no research has been conducted so far on which f is the best for obtaining high sensitivity. In the present invention, as described in detail below, the results of this research under actual conditions found that the smaller f is, the higher the overall light-gathering efficiency is. This means that the effect of increasing the amount of light detected from the center of the light-emitting point by reducing f is greater than the effect of increasing the amount of light detected from the end of the light-emitting point by increasing f.

[0020] On the other hand, crosstalk requires a different discussion from the above-mentioned light-gathering efficiency. If f becomes smaller, the above-mentioned angle θ becomes larger, and thus the ratio of the light emitted from the end of the light-emitting point and the adjacent sensor (or the area to which a part of the area sensor is allocated) configured to detect the light from the adjacent light-emitting point increases, thereby increasing the crosstalk in detecting the light from the adjacent light-emitting point. That is, it can be seen that there is a trade-off between reducing f to improve the light-gathering efficiency and thus obtain high sensitivity, and increasing f to reduce crosstalk. However, including Patent Document 2, no research has been conducted so far on which f is the best in order to achieve both high sensitivity and low crosstalk.

[0021] Solutions to Solve Problems

[0022] The luminescence detection device of the present invention comprises: a focusing lens array, in which M focusing lenses are arranged, and the M focusing lenses individually focus the light from the luminescence point array in which M luminescence points are arranged to form light beams, wherein M≥2; and at least one sensor, which causes the M light beams to enter in parallel without focusing them, and sets the average value of the effective diameters of the M luminescence points to d, the average value of the focal lengths of the M focusing lenses to f, the average value of the intervals between the M focusing lenses to p, and the average value of the maximum optical path lengths between the M focusing lenses and the sensor to g. At this time, d, f, p, and g satisfy a predetermined prescribed relationship so that the M light beams can be detected with low crosstalk or high sensitivity.

[0023] The effects of the invention are as follows.

[0024] According to the present invention, a device for detecting light from a light-emitting point array with high sensitivity and low crosstalk can be made small, and the overall size of various devices using the detection device can be made small. Therefore, the space for placing the device can be reduced, and the device can be carried, thereby improving the convenience of using the device. In addition, by reducing the number of components constituting the device and making the components themselves small, the manufacturing cost can be reduced.

[0025] Other problems, structures, and effects than those described above will become apparent from the following description of the embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a schematic diagram of a structure in which a focusing lens is used to focus light from a light-emitting point and form it into a light beam.

[0027] Figure 2 This diagram shows the relationship between the optical path length g between the condenser lens and the sensor and the relative amount of detected light, using the focal length f of the condenser lens as a parameter.

[0028] Figure 3 This is a schematic diagram of a structure in which light emitted from two adjacent light-emitting points is individually condensed by condensing lenses to form separate light beams.

[0029] Figure 4 This is a schematic diagram of a structure in which light emitted from two adjacent light-emitting points is individually condensed by condensing lenses to form a mixed light beam.

[0030] Figure 5 1 is a diagram showing the relationship between the optical path length g between the condenser lens and the sensor and the focal length f of the condenser lens, which satisfies the conditions of high sensitivity and low crosstalk.

[0031] Figure 6 This is a schematic diagram of a light emission detection device that uses a condenser lens array to individually condense light emitted from a light-emitting point array so that the light is incident in parallel on a color sensor for detection.

[0032] Figure 7 This is a schematic diagram of a luminescence detection device that uses a focusing lens array to individually focus the light from a luminous point array, uses a dichroic mirror array to split the light beams into different wavelength bands in parallel, and makes them enter a sensor in parallel for detection.

[0033] Figure 8 It is a schematic diagram of a luminescence detection device that individually uses a condenser lens array to condense light from a light-emitting point array, uses a wavelength dispersion element to disperse the above light beams in parallel, and uses an imaging lens array to individually image and detect.

[0034] Fig. 9 This is a schematic diagram of a light emission detection device that individually uses a focusing lens array to focus light from a light-emitting point array, uses an optical element to deflect the light beams in parallel, and causes them to enter a sensor in parallel for detection.

[0035] Fig.10 This is a schematic diagram of a luminescence detection device that individually uses a focusing lens array to focus light from a light-emitting point array, uses an optical fiber array to deflect the light beams in parallel, and causes the light beams emitted from the optical fiber array to enter a sensor in parallel for detection.

[0036] Fig.11 It is a schematic diagram showing an example of the device structure of a capillary array DNA sequencer.

[0037] Fig.12 This is a diagram showing the result of calculating the maximum width of parallel light beams that can be split in an example of a dichroic mirror array that splits light beams incident parallel to the dichroic mirror array in the vertical direction.

[0038] Fig.13 This is a diagram showing the result of calculating the maximum width of parallel beams that can be split in an example of a dichroic mirror array that splits beams incident perpendicularly to the dichroic mirror array in the same direction.

[0039] Fig.14 This is a diagram showing the result of calculating the maximum width of parallel beams that can be split in an example of a dichroic mirror array that splits beams incident perpendicularly to the dichroic mirror array in the same direction and becomes compact.

[0040] Fig.15 The diagram shows the result of calculating the maximum width of parallel beams that can be split in an example of a dichroic mirror array that is compact and has a step difference and splits beams incident perpendicularly to the dichroic mirror array in the same direction.

[0041] Fig.16 This is a diagram showing the structure of a dichroic mirror array in an optimal mode for shortening the optical path length and expanding the opening width.

[0042] Fig.17 is the optical path length of the dichroic mirror array, L max Below, the opening diameter is W min A diagram showing the relationship between the thickness β of the dichroic mirror and the interval x of the dichroic mirror array in the above case.

[0043] Fig.18 3 is a diagram showing the relationship between the interval x of the dichroic mirror array, the opening width W, and the optical path length change ΔL.

[0044] Fig.19 1 is a diagram showing the relationship between the step differences y and z and the opening width W of the dichroic mirror array.

[0045] Fig. 20 The diagram shows the result of calculating the maximum splittable width of parallel beams in an example in which a dichroic mirror array is split in the same direction, incident perpendicularly to the dichroic mirror array and the dichroic mirrors are tilted more than 45 degrees with respect to the same direction.

[0046] Fig.21 1 is a diagram showing the relationship between the inclination θ0 of the dichroic mirror and the opening width W of the dichroic mirror array.

[0047] Fig. 22 This is a schematic diagram of a light emission detection device that allows part of the light emitted from a light-emitting point array to individually transmit through a pinhole array, and the transmitted light is individually condensed by a condenser lens array and incident in parallel on a sensor for detection.

[0048] Fig.23 This is a schematic diagram of a luminescence detection device that allows part of the light emitted from an array of light-emitting points arranged at intervals smaller than a focusing lens array to individually transmit a pinhole array, and the transmitted light is individually focused by a focusing lens array and incident in parallel on a sensor for detection.

[0049] Fig.24 This is a schematic diagram of a luminescence detection device that allows part of the light emitted from a luminous point array to individually pass through pinholes and individually focus the transmitted light using a focusing lens array, uses a color dichroic mirror array to split the resulting light beams into different wavelength bands in parallel, and allows them to be incident on a sensor in parallel through an optical path length adjustment element for detection.

[0050] Fig.25 It is a schematic diagram of forming an image of a light emitting area of ​​a light emitting detection device including a pinhole array, a focusing lens array, and a scanning mechanism.

[0051] Fig.26 This is a schematic cross-sectional view showing a structural example of a device in which a V-groove array in which a plurality of capillaries are arranged and an individual condenser lens array are integrated.

[0052] Fig. 27 This is a schematic cross-sectional view showing a structural example of a device in which a V-groove array in which a plurality of capillaries are arranged, a pinhole array, and an individual condenser lens array are integrated.

[0053] Fig.28 The present invention is a schematic cross-sectional view showing a structural example of an opaque device in which a V-groove array in which a plurality of capillaries are arranged and a pinhole array are integrated, and a device in which a separate condenser lens array is bonded and integrated. DETAILED DESCRIPTION

[0054] Figure 1This is a cross-sectional schematic diagram including an optical axis of an optical system that uses a condenser lens 2 to condense the light emitted from a light-emitting point 1 and detects it using a sensor arranged at a certain distance from the condenser lens. In the present invention, the terms such as light-emitting point and light emission are often used, but they are not limited to the case where the substance as the detection object itself emits fluorescence, phosphorescence, etc., and there are also cases where scattered light generated from the detection object as a result of irradiating light to the detection object and transmitted light after passing through the detection object are expressed together as light emission from the light-emitting point.

[0055] The diameter of the light-emitting point 1 is d, the focal length of the condenser lens 2 is f, the effective diameter of the condenser lens 2 is D, the diameter of the detection area of ​​the sensor is D, and the optical distance (optical path length) between the condenser lens 2 and the sensor is g. If the center of the light-emitting point 1 is arranged at a distance f between the light-emitting point 1 and the condenser lens 2, that is, at the focal point of the condenser lens 2, the light emitted from the center of the light-emitting point 1 becomes a parallel light beam 3 with a diameter of D due to the condenser lens 2 and travels along the optical axis. In the sensor at a position away from the condenser lens 2 by an optical path length of g, the parallel light beam 3 forms a light spot 4 with a diameter of D. Figure 1 The lower side of the optical system shows a light spot 1 observed from the optical axis direction, and the upper side shows a light spot 4 observed from the optical axis direction and a light spot 5 described later. On the other hand, the light emitted from the left end of the light spot 1 becomes a parallel light beam 3' with a diameter D due to the condenser lens 2, and travels in a direction forming an angle θ with the optical axis. In the sensor, the parallel light beam 3' forms a light spot 5 with a diameter D.

[0056] The condensing efficiency of the light emitted from the center of the light emitting point 1 is calculated by using the F value of the condensing lens 2 and summing it with 1 / F 2 Since F = f / D, if D is set to be constant, the smaller f is, the greater the light collection rate is. On the other hand, the light spot 5 from the left end of the light spot 1 deviates from the detection area of ​​the sensor to the right. In other words, the light spot 4 is detected by the sensor in its entirety, but the light spot 5 is detected only at a ratio that overlaps with the light spot 4. The greater the overlap, the greater the amount of light detected in the entire area of ​​the light spot. For this reason, the angle θ formed with the optical axis of the parallel light beam 3' can be smaller, since θ = tan -1 (d / 2 / f), so if d is set constant, the larger f is, the better. As described above, in order to increase the detection light amount of the light-emitting point 1, there is a trade-off relationship between reducing f and increasing f, but no research has been conducted so far on which f is the best. Therefore, the conditions of f and g for increasing the detection light amount of the light-emitting point 1 are described below.

[0057] In order to evaluate the detection light quantity, the Figure 3The fluorescence detection device shown in FIG. 1 is used as a reference. In a typical example of the fluorescence detection device, the focal length of the common condenser lens is f1=50mm, and the effective diameter D1≥25mm. The brightness of the lens is F=f1 / D1≤2.0. Therefore, when a condenser lens with F0=2.0 is used, the light from the infinitesimal-sized light-emitting point located at the focal point becomes a parallel light beam due to the lens, and when all its light is detected by the sensor without loss, the detected light amount is used as the reference (100%). In the following, the detected light amount of any infinitesimal-sized light-emitting point is evaluated according to the relative detected light amount with respect to the above-mentioned reference. In addition, it is considered that the finite-sized light-emitting point with an average effective diameter d is composed of a plurality of infinitesimal-sized light-emitting points. In this specification, "finite-sized light-emitting point" is simply referred to as "light-emitting point", and "infinitely small-sized light-emitting point" is referred to as "infinitely small-sized light-emitting point" each time. The relative detected light amount of a light-emitting point is the average value of the relative detected light amounts of the plurality of infinitesimal-sized light-emitting points. For example, in the above example, if the focusing lens is replaced from F = 2.0 to F = 1.4, the focusing efficiency becomes (F0 / F) 2 =2.0 times, so that the relative detection light amount of the infinitesimal size light point becomes 200%. However, it is assumed that the total light amount emitted from the light point in all directions is constant, and the light density inside the light point is spatially uniform. In addition, in a typical example of the present fluorescence detection device, the interval p of the light point array is 0.36 mm, the number of light points is M = 24, and the full width AW of the light point array is p × (M-1) = 8.28 mm. Since the light point located in the center of the light point array is located near the focus of the lens, the relative detection light amount is approximately 100%, but since the light point located at the end of the light point array is far from the focus of the lens, the relative detection light amount is reduced to about 50%. Therefore, in the present invention, the goal is as follows: the relative detection light amount of each light point becomes more than 50%, and the detection sensitivity of each light point is the same as or higher than before.

[0058] Figure 2 is Figure 1 The figure shows the result of calculating the relationship between g and the relative detection light quantity using f as a parameter in the structure shown. Here, the average effective diameter of the light-emitting point 1 is set to d = 0.05 mm. In addition, the effective diameter of the individual focusing lens 2 is set to D = 0.5 mm. The relative detection light quantity is calculated by considering the brightness of the lens F = f / 0.5. The light-emitting point 1 with an effective diameter of d = 0.05 mm is composed of about 500 infinitesimal light-emitting points with an interval of 0.1 μm. In each infinitesimal light-emitting point, according to Figure 1 The relative detection light quantity is calculated based on the same idea as the overlapping area ratio of light spot 4 and light spot 5, and the relative detection light quantity of light spot 1 is obtained based on the average value of the above values. Figure 2As shown, it was found for the first time that the smaller f and the smaller g are, the larger the relative detection light quantity is. This shows that the effect of increasing the relative detection light quantity of the infinitesimal light emitting point located at the center of the light emitting point 1 by reducing f is greater than the effect of increasing the above-mentioned overlapping area ratio by increasing f. In addition, it is shown that for any f, the effect of increasing the above-mentioned overlapping area ratio by reducing g is greater.

[0059] Figure 1 as well as Figure 2 In the embodiment, the center of the light-emitting point 1 is arranged at the focal point of the condenser lens 2, so that the light is emitted as a parallel light beam. The present invention also functions well under this condition, but as described in detail below, if the center of the light-emitting point 1 is slightly away from the focal point of the condenser lens 2, so that the light is slightly reduced as a light beam 6, both high sensitivity and low crosstalk can be achieved, and it can be known that it becomes a more suitable condition. More specifically, when the image of the light-emitting point 1 is connected at the sensor position, that is, at the position of the optical path length g from the condenser lens 2, the diameter of the light-emitting point 1 can be minimized, so it can be known that it is the best condition.

[0060] Figure 3 This is a cross-sectional view including the optical axis of an optical system in which the light emitted from two adjacent light-emitting points 1 is respectively concentrated by individual condensing lenses 2 and light-emitting point images 7 as images of the light-emitting points 1 are obtained at the sensor position. In the present invention, the expression "light-emitting point image" does not necessarily refer to an image in which the light emitted from the light-emitting point is imaged, but generally refers to a cross section at a predetermined position in which the light beam emitted from the light-emitting point is concentrated. The diameter of the light-emitting point 1 is d, the focal length of the condensing lens 2 is f, the effective diameter of the condensing lens 2 is D, the interval between the light-emitting points 1 and the interval between the condensing lenses 2 are p, the diameter of the detection area of ​​the sensor is D, the optical distance (optical path length) between the condensing lens 2 and the sensor is g, and the diameter of the light-emitting point image 7 at the sensor position is d'. When the distance between the light-emitting point 1 and the condensing lens 2 is adjusted and the light emitted from the light-emitting point 1 is imaged at the sensor position by the condensing lens 2, the diameter d' of the light-emitting point image 7 becomes minimum.

[0061] At this time, the image magnification is expressed as follows,

[0062] [Formula 1]

[0063] m=(g-f) / f

[0064] The diameter d' of the luminous point image 7 is expressed as follows.

[0065] [Formula 2]

[0066] d'≥m×d=(g-f) / f×d

[0067] Here, the equation (2) holds true when the light emitted from the light emitting point 1 is imaged at the sensor position by the condenser lens 2 , and holds true otherwise. Figure 3 The lower side of the optical system shows the light-emitting point 1 observed from the optical axis direction, and the upper side shows the light-emitting point image 7 observed from the optical axis direction. The present invention works well when g ≥ 2 × f, that is, when m ≥ 1. More preferably, m ≥ 5, and a further good condition is m ≥ 10. This is because, from Figure 2 It is clear that in order to increase the relative detection light quantity, f needs to be reduced to the mm level, but it is physically difficult to reduce g in this way.

[0068] In the various figures of this specification, the light-emitting point 1 and the light-emitting point image 7 are depicted by circles, but in reality they are not limited to circles and may have other shapes. Generally, the diameter d of the light-emitting point 1 and the diameter d' of the light-emitting point image 7 are the widths of the light-emitting point 1 and the light-emitting point image 7 in the arrangement direction, respectively. In addition, as described below, in the same light-emitting detection device, there are sometimes multiple optical distances (optical path lengths) between the condenser lens 2 and the sensor. In this case, the optical distance between the condenser lens 2 and the sensor is generally taken as the optical path length s, and the maximum value of s is taken as the optical path length g, so that the above-mentioned equations (1), (2) and the following equations (3) to (18) are established. The reason for this is described below. If the optical distance between the condenser lens 2 and the sensor is expressed by the variable s, g is taken as the maximum value, and 0≤s≤g. At this time, the diameter d'(s) of the light-emitting point image 7 at any s is smaller than the larger one of d'(s=0)=D and d'(s=g)=d'. That is, when D≥d', d'(s)≤D, and when D≤d', d'(s)≤d'. In the former case, no crosstalk occurs, and in the latter case, if the crosstalk reference can be cleared when s=g, it means that the reference can also be cleared when 0≤s≤g. On the other hand, the relative detection light amount is constant and does not change with s.

[0069] Furthermore, in this specification, the focusing lens 2 is basically a circle whose diameter is the effective diameter D, but this is not necessarily the case. Generally, the effective diameter D of the focusing lens 2 represents the width in the arrangement direction of the light-emitting points 1 and the arrangement direction of the focusing lens 1, and the width in the direction perpendicular to the arrangement direction is not the same. The focusing lens 2 can be circular, elliptical, square, rectangular, or can be other shapes. In the above discussion, it is considered that the diameter d'(s=0)=D of the light-emitting point image 7 is the diameter in the arrangement direction of the light-emitting points 1 and the arrangement direction of the focusing lens 1. No matter how large the diameter of the light-emitting point image 7 in the direction perpendicular to the arrangement direction is, it will not be conducive to crosstalk. Moreover, if g is sufficiently large, d'(s=g)=d' has nothing to do with D. Therefore, the conditions of the following equations (13) to (18) related to crosstalk are independent of the width of the focusing lens 2 in the direction perpendicular to the arrangement direction and remain unchanged. On the other hand, if the width of the focusing lens 2 in the direction perpendicular to the arrangement direction is larger than the effective diameter D, the F value may be smaller than F=f / D, that is, the focusing efficiency may be higher. In this case, the following conditions of formula (3) to formula (12) related to sensitivity can bring about a further higher relative detection light amount and sensitivity.

[0070] First, the conditions for obtaining high sensitivity are studied. The focusing efficiency of the focusing lens 2 of the light from the light emitting point 1 can be expressed by the F value of the focusing lens 2, F=f / D. In order to make the relative detection light amount more than 50%, it is set to F≤2.8, that is, f≤2.8×D. On the other hand, in order to form a focusing lens array, it is necessary to set p≥D, so the following formula (3) is the condition for the relative detection light amount to be more than 50%.

[0071] [Formula 3]

[0072] f≤2.8×p

[0073] Similarly, in order to make the relative detection light amount greater than 100%, greater than 200%, greater than 400% and greater than 800%, F≤2.0, 1.4, 1.0 and 0.7 respectively, that is, the following equations (4), (5), (6) and (7) are the conditions.

[0074] [Formula 4]

[0075] f≤2.0×p

[0076] [Formula 5]

[0077] f≤1.4×p

[0078] [Formula 6]

[0079] f≤1.0×p

[0080] [Formula 7]

[0081] f≤0.7×p

[0082] The above equations (3) to (7) are correct when the distance between the light-emitting point 1 and the focusing lens 2 can be approximated as f, but more strictly speaking, they can be expressed as follows. The distance between the light-emitting point 1 and the focusing lens 2 is f when the light from the light-emitting point 1 is imaged at the optical distance g by the focusing lens 2. 2 / (g-f)+f, so the effective F value of the condenser lens 2 can be expressed as F'=(f 2 / (g-f)+f) / D. Therefore, in order to make the relative detection light amount 50%, 100% or more, 200% or more, 400% or more, and 800% or more, the following equations (8), (9), (10), (11), and (12) are strict conditions.

[0083] [Formula 8]

[0084] f≤(1 / (2.8×p)+1 / g) -1

[0085] [Formula 9]

[0086] f≤(1 / (2.0×p)+1 / g) -1

[0087] [Formula 10]

[0088] f≤(1 / (1.4×p)+1 / g) -1

[0089] [Formula 11]

[0090] f≤(1 / (1.0×p)+1 / g) -1

[0091] [Formula 12]

[0092] f≤(1 / (0.7×p)+1 / g) -1

[0093] Next, we examine the conditions for achieving low crosstalk. Figure 3 As shown, when the luminous point images 7 of adjacent luminous points 1 do not overlap each other, there is no crosstalk. Figure 4 As shown in FIG. 1 , the luminous point images 7 of adjacent luminous points 1 overlap with each other, thus generating crosstalk. Figure 4 The crosstalk is expressed as a ratio X of the overlapping area of ​​adjacent luminous point images 7 to the area of ​​the luminous point images 7. To make the crosstalk equal to or less than X, the following equation (13) is used, and the following equation (14) becomes a condition.

[0094] [Formula 13]

[0095] X=1 / π×(cos -1 (V 2 / 2-1)-sin(cos -1 (V 2 / 2-1)))

[0096] [Formula 14]

[0097] V≤2×p / d'

[0098] If equation (2) is used to transform equation (14), it can be expressed as follows.

[0099] [Formula 15]

[0100] f≥1 / ((2×p) / (V×d)+1)×g

[0101] In order to detect the light emitted from the light emitting point 1 to be detected without being affected by the light emitted from the light emitting points 1 on both sides, Figure 4 In the equation (13), the distance between the two luminous point images 7 needs to be at least greater than the radius (or half of the diameter) of the luminous point image. If expressed by equations (13) and (14), X = 0.39 (39%), V = 1, and equation (15) can be expressed as follows.

[0102] [Formula 16]

[0103] f≥1 / (2×p / d+1)×g

[0104] In order to more effectively and independently detect the light emission from multiple light-emitting points 1, it is preferred to make the total proportion of crosstalk from both sides less than 50%. Therefore, if expressed by formula (13) and formula (14), X = 0.25 (25%), V = 1.27. For formula (15), the following formula (17) is the condition.

[0105] [Formula 17]

[0106] f≥1 / ((2×p) / (1.27×d)+1)×g

[0107] It is further preferred to make the crosstalk 0%. Therefore, if it is expressed by equations (13) and (14), X = 0 (0%), V = 2. For equation (15), the following equation (18) is the condition:

[0108] [Formula 18]

[0109] f≥1 / (p / d+1)×g

[0110] As described above, for given p and d, by selecting g and f that satisfy any one of equations (3) to (12), the desired relative detection light quantity and sensitivity can be obtained. On the other hand, for given p and d, by selecting g and f that satisfy any one of equations (16) to (18), the desired crosstalk can be obtained. That is, by selecting g and f that satisfy any one of equations (3) to (12) and any one of equations (16) to (18), the relative detection light quantity and crosstalk in a compromise relationship can be obtained at a desired level.

[0111] Figure 5 As a typical example, the horizontal axis g and the vertical axis f show equations (3) to (12) and equations (16) to (18) when p = 1 mm and d = 0.05 mm. The numbers shown in the curves or straight lines show the boundary of the equations with the corresponding numbers, ↓ shows the area below the boundary, and ↑ shows the area above the boundary. For example, in order to satisfy the condition that the relative detection light amount is 50% or more in equation (3), Figure 5 On the other hand, in order to satisfy the condition of making the crosstalk less than 25% in equation (17), the g and f in the area below the straight line ↓(3) are sufficient. Figure 5 That is, in order to make the relative detection light amount more than 50% and the crosstalk less than 25%, it is necessary to Figure 5 The g and f in the area below the straight line ↓(3) and above the straight line ↑(17) are sufficient.

[0112] For those who use Figure 5 As for the luminescence detection device of g and f shown in the figure, it can be clearly seen from the sizes of g and f that it not only has the performance of high sensitivity and low crosstalk, but also has the feature of being able to reduce the size of the device by one to three digits compared with the detection devices of Patent Document 1 and Non-Patent Document 1. It can be clearly seen from the above content that the smaller p is and the larger d is, the narrower the range of g and f that meet the conditions of high sensitivity and low crosstalk is. On the other hand, the miniaturization of the luminescence detection device becomes an inevitable structure. In other words, the smaller p is and the larger d is, the more significant the effect is when the features of the present invention are flexibly utilized compared with the existing methods. Specifically, the features of the present invention are particularly flexibly utilized when p≤20mm and more preferably p≤10mm. Moreover, the features of the present invention are particularly flexibly utilized when d≥0.005mm and more preferably d≥0.01mm.

[0113] Next, based on the above conditions, the method of performing multicolor detection is further described. Figure 3 or Figure 4The color sensor is configured in such a manner that the sensor surface is perpendicular to the optical axis of the focusing lens 2, that is, the sensor surface is parallel to the arrangement plane of the light-emitting points 1 and the arrangement plane of the focusing lens 2, according to the position of the light-emitting point image 7 in the light-emitting point image 7. Here, the color sensor is arranged with two or more pixels that can respectively identify and detect at least two or more different wavelengths of light. The most common color sensor is a color sensor used in a digital camera for ordinary consumers, in which a plurality of three pixels that identify the three colors of R, G, and B, namely red, green, and blue, are arranged in two dimensions. In the present invention, such an ordinary color sensor can also be used. The sensitivity of ordinary color sensors in recent years is relatively high, so they can also be used in the technical field. The above-mentioned color sensor is most suitable for the identification of three colors, but as performed in a digital camera, it is also possible to identify four or more colors based on the intensity ratio of the three pixels. Therefore, the detection device using the above-mentioned color camera can also be applied to a DNA sequencer that performs four-color detection.

[0114] However, in order to accurately identify the color of the light emitted from the light-emitting point 1, it is effective to make the diameter d' of the light-emitting point image 7 larger than the size of various pixels. This is because, for one light-emitting point 1 and various pixels, by using multiple pixels to detect light emission, the influence of the deviation of the relative position of the light-emitting point image 7 and the pixel can be avoided. Among the suitable conditions of the present invention shown in equations (3) to (12) and equations (16) to (18), there are many cases where m>1 in equation (1) and d'>d in equation (2), so that the above conditions are easily satisfied. In addition, since the arrangement plane of the light-emitting point 1 and the arrangement plane of the focusing lens 2 are parallel to the sensor surface, the sensor surface can be made close to the arrangement plane of the focusing lens 2, so that it is easy to satisfy the conditions based on equations (3) to (12) and equations (16) to (18).

[0115] Recently, color sensors with four kinds of pixels, in addition to R, G, and B, are sold on the market. It is advantageous to use such color sensors for the detection of four colors of DNA sequencers. A variety of pixels can be arranged on the same plane or in a direction perpendicular to the sensor surface. As described above, applying a color sensor that has been actually used to the present invention is effective in suppressing development costs. Of course, the number of types of pixels of the color sensor and the characteristics of the colors recognized by the various pixels can also be customized according to the purpose.

[0116] Figure 6 An example of a multi-color detection device using a color sensor is shown. Figure 6 (a) shows the multicolor detection device viewed from a direction perpendicular to a plane including the optical axes of the condenser lens 2, Figure 6(b) shows an image 12 detected by the two-dimensional color sensor 11. Here, an example of performing four-color detection is shown.

[0117] like Figure 6 As shown in (a), the light from each light emitting point 1 is individually condensed by a condenser lens 2 to form a light beam 9, which is then transmitted in parallel through a common long-pass filter 10 and then incident in parallel on a common two-dimensional color sensor 11. The long-pass filter 10 is configured to block light of a wavelength that hinders multi-color detection, such as excitation light. Figure 6 As shown in (b), a luminous point image 7 of the light emitted from each luminous point 1 is formed on the image 12 of the two-dimensional color sensor 11. The two-dimensional color sensor 11 is configured to regularly arrange a plurality of four types of pixels, for example, A pixels 13 for mainly detecting A light, B pixels 14 for mainly detecting B light, C pixels 15 for mainly detecting C light, and D pixels 16 for mainly detecting D light. The size of each pixel 13, 14, 15, 16 is S = 0.05 mm. On the other hand, if d = 0.05 mm, f = 1 mm, g = 10 mm, and the luminous point 1 is imaged on the two-dimensional color sensor, then m = 9 is obtained from equation (1), and d' = 0.45 mm is obtained from equation (2). That is, if S < d' is satisfied, the luminous point image 7 is detected by about 64 pixels, and each type of pixel is detected by about 16 pixels.

[0118] In this way, for each pixel, by detecting each luminous point image 7 with multiple pixels, it is possible to detect the four colors of light emitted from the luminous point 1 with high accuracy. For example, even if the relative position of each pixel and the luminous point image 7 changes, there is no problem. Or, even if the light intensity distribution in the luminous point image 7 is uneven, each color can be detected evenly and stably. Figure 6 In the multi-color detection device shown, the diameter d of each light-emitting point 1, the interval p between each light-emitting point 1 and each focusing lens 2, the focal length f of each focusing lens 2, the effective diameter D, and the optical distance g between each focusing lens 2 and the sensor 11 satisfy any one of Formulas (3) to (7), (8) to (12), and (16) to (18), thereby achieving the predetermined high sensitivity and low crosstalk, and realizing the miniaturization and low cost of the detection device.

[0119] On the other hand, in Figure 6 When a color sensor having multiple types of pixels arranged on the same plane is used as shown, there is a problem of low efficiency in utilizing incident light. Figure 6As shown in FIG. 1 , when a color sensor having four types of pixels arranged therein is used to identify four colors, the utilization efficiency of the incident light is less than 1 / 4. This sometimes becomes an obstacle to the case of performing luminescence detection with higher sensitivity. It is possible to improve the utilization efficiency of the incident light by using a color sensor having multiple types of pixels arranged in a direction perpendicular to the sensor surface, but such a color sensor is not yet commonly used. Therefore, next, another method of multi-color detection with higher utilization efficiency of the incident light is proposed.

[0120] One of the schemes for improving the utilization efficiency of incident light is to use more than one color separation mirror (hereinafter referred to as color separation mirror). The color separation mirror forms a multilayer film on the front of at least one side of a transparent substrate such as glass to make the reflected light and the transmitted light of the light generally incident at 45° into light of different wavelength bands. By flexibly utilizing both the reflected light and the transmitted light, the utilization efficiency of the incident light can be improved. Generally, if one color separation mirror is used, a maximum of two colors can be detected. If two color separation mirrors are combined, a maximum of three colors can be detected. Similarly, if N color separation mirrors are combined, a maximum of N+1 colors can be detected. In addition to the above-mentioned color separation mirrors, there are also many cases where bandpass filters, colored glass filters, or total reflection mirrors are used at the same time. In digital cameras for ordinary consumers, there is a device that combines three color separation mirrors and uses three CCDs to perform three color detections. In addition, non-patent document 1 is an example of combining three color separation mirrors and using four CCDs to perform four color detections. In this way, in the method of using the color separation mirror, since the forward directions of the reflected light and the transmitted light are different, there are many cases where multiple sensors are used. This goes against the purpose of the present invention, which is to reduce the size and cost of the detection device.

[0121] Figure 7 An example of a multicolor detection device that solves these problems is shown. Figure 7 (a) shows the multicolor detection device viewed from a direction perpendicular to a plane including the optical axes of the condenser lens 2, Figure 7 (b) shows a cross section of a multicolor detection device including the optical axis of a condenser lens 2 and perpendicular to the arrangement direction of the condenser lens array, Figure 7 (c) shows the image 29 detected by the two-dimensional sensor 30. Here, an example of performing four-color detection is shown.

[0122] As an example, the light from each light point 1 of the four light point arrays is focused by a separate focusing lens 2 to form a light beam 9, which is then transmitted in parallel through a common long pass filter 10. Figure 6 Here, use Figure 7(b) shows a dichroic mirror array in which four common dichroic mirrors 17, 18, 19 and 20 are arranged. Each light beam 9 is incident on each dichroic mirror 17 to 20 in parallel, and is split into four light beams 21, 22, 23 and 24 in a direction perpendicular to the light-emitting point array direction, and these light beams are respectively directed in the same direction as the light beam 9, that is, in the direction of the optical axis of the condenser lens 2, and are incident on a common two-dimensional sensor 30 in parallel, thereby forming light-emitting point images 25, 26, 27 and 28. Here, the dichroic mirror 20 may be replaced by a total reflection mirror, but is hereinafter referred to as the dichroic mirror 28 for simplicity.

[0123] like Figure 7 As shown in (c), 16 light-emitting point images 25 to 28, which are obtained by dividing the light from the four light-emitting points 1 into four parts, are observed together on the image 29 of the two-dimensional sensor 30. Here, in the light beam 9, the light beam that passes through the dichroic mirror 17 is the light beam 21, the light beam that is reflected by the dichroic mirrors 17 and 18 is the light beam 26, the light beam that is reflected by the dichroic mirror 17, passes through the dichroic mirror 18, and then is reflected by the dichroic mirror 19 is the light beam 23, and the light beam that is reflected by the dichroic mirror 17, passes through the dichroic mirrors 18 and 19, and then is reflected by the dichroic mirror 20 is the light beam 24. By designing and controlling the transmission characteristics and reflection characteristics of the long-pass filter 10 and the dichroic mirrors 17 to 20, the light beam 21 mainly has the A fluorescence component, the light beam 22 mainly has the B fluorescence component, the light beam 23 mainly has the C fluorescence component, and the light beam 24 mainly has the D fluorescence component, and by detecting the intensities of the light-emitting point images 25, 26, 27, and 28, the A, B, C, and D fluorescence can be detected.

[0124] The wavelengths of the light beams 21, 22, 23, and 24 can be arbitrarily designed, but if they are arranged in order of wavelength, the design of the dichroic mirrors 17 to 20 will be easy. That is, the central wavelength of the A fluorescence can be set to be greater than the central wavelength of the B fluorescence, greater than the central wavelength of the C fluorescence, and greater than the central wavelength of the D fluorescence, or the central wavelength of the A fluorescence can be set to be less than the central wavelength of the B fluorescence, less than the central wavelength of the C fluorescence, and less than the central wavelength of the D fluorescence. Figure 7 Although not shown in the figure, it is effective to arrange a bandpass filter or a colored glass filter having different spectral characteristics at at least one position of the light beams 21, 22, 23 and 24 to compensate for or improve the spectral characteristics of the dichroic mirrors 17 to 20. Figure 7Although not shown, there may also be irradiation light such as excitation light for causing the light-emitting point 1 to emit light. If such irradiation light is irradiated from a direction perpendicular to the optical axis of the condensing lens 2 without using the condensing lens 2, the ratio of the irradiation light entering the sensor through the condensing lens 2 is reduced, which is beneficial to sensitivity. In addition, a so-called incident luminescence detection structure may be formed as follows to replace the long-pass filter 10: a dichroic mirror independent of the dichroic mirrors 17 to 20 is configured, the irradiation light is reflected by the above-mentioned dichroic mirror, and then focused by the condensing lens 2 and irradiated to the light-emitting point 1, the light from the light-emitting point 1 is focused by the condensing lens 2, and then passes through the above-mentioned dichroic mirror, and is detected by the condensing lens 2. Figure 7 The same multi-color detection device was used for detection.

[0125] exist Figure 7 In the multicolor detection device shown, the diameter d of each light-emitting point 1, the interval p between each light-emitting point 1 and each focusing lens 2, the focal length f of each focusing lens 2, the effective diameter D, and the optical distance g between each focusing lens 2 and the sensor 30 satisfy any one of equations (3) to (7), (8) to (12), and (16) to (18), thereby achieving the specified high sensitivity and low crosstalk, and achieving miniaturization and low cost of the detection device. Here, the following (1) to (10) summarize the implementation of the use Figure 7 The dichroic mirror array shown has the characteristics of miniaturization and cost reduction of the multi-color detection device. It is not necessary to satisfy all of these characteristics, but satisfying any one of them is effective.

[0126] (1) For the M light-emitting points of the light-emitting point array, the M light beams emitted from each light-emitting point are focused by the focusing lens array and split into N light beams having different wavelength components, and these light beams are made to travel in the same direction.

[0127] (2) For the M light-emitting points of the light-emitting point array, the M light beams emitted from each light-emitting point are focused by the focusing lens array and split into N light beams with different wavelength components, and these light beams are made to travel along the optical axis direction of each focusing lens.

[0128] (3) For the M light-emitting points of the light-emitting point array, the direction in which the M light beams emitted from each light-emitting point are focused by the focusing lens array into different wavelength components is perpendicular to the arrangement direction of the light-emitting point array and the focusing lens array.

[0129] (4) For the M light-emitting points of the light-emitting point array, the directions in which the M light beams emitted from each light-emitting point are focused by the focusing lens array into different wavelength components are perpendicular to the optical axes of the focusing lenses.

[0130] (5) For the M light-emitting points of the light-emitting point array, N color dichroic mirrors divide the M light beams emitted from each light-emitting point into different wavelength components after being focused by the focusing lens array. The N color dichroic mirrors are arranged in a direction perpendicular to the arrangement direction of the light-emitting point array and the focusing lens array.

[0131] (6) For the M light-emitting points of the light-emitting point array, N dichroic mirrors separate the M light beams emitted from each light-emitting point into different wavelength components after being focused by the focusing lens array. The N dichroic mirrors are arranged in a direction perpendicular to the optical axis of each focusing lens.

[0132] (7) With respect to the M light-emitting points of the light-emitting point array, the M light beams obtained by focusing the light from each light-emitting point by the focusing lens array are respectively divided into N different wavelength components, thereby obtaining M×N light beams, so that the M×N light beams are no longer focused but directly incident on the sensor.

[0133] (8) With respect to the M light-emitting points of the light-emitting point array, the optical axis of each condenser lens of the condenser lens array for condensing light emitted from each light-emitting point is perpendicular to the sensor surface.

[0134] (9) The system is composed of N different types of dichroic mirrors, each of which is composed of a single component, so that M light beams obtained by individually focusing light emitted from M light-emitting points of the light-emitting point array are incident in parallel on each dichroic mirror.

[0135] (10) M light beams obtained by individually focusing light emitted from M light-emitting points of a light-emitting point array are split into N different wavelength components to obtain M×N light beams, which are then incident in parallel on a single sensor.

[0136] In the above, the following case is described: for each light beam formed by each light from the light-emitting point array being focused by each condenser lens of the condenser lens array, it is not re-condensed by other lenses other than the condenser lens, but directly incident on the sensor. In the following, the following case is described: for each light beam formed by each light from the light-emitting point array being focused by each condenser lens of the condenser lens array, it is re-condensed by other lenses other than the condenser lens, and then incident on the sensor. In the following, the lens for re-condensing each light beam is referred to as a re-condenser lens.

[0137] When deriving equations (1) to (18), the optical distance between the focusing lens and the sensor is set to g. However, when using a re-focusing lens, if the optical distance between the focusing lens and the re-focusing lens is set to g, equations (1) to (18) can remain unchanged to achieve high sensitivity and low crosstalk conditions. In other words, Figure 3 or Figure 4In the figure, it is considered that the position of the luminous point image 7 is not the position of the sensor but the position of the re-focusing lens. This is because, assuming that the conditions of high sensitivity and low crosstalk are not met at the position of the re-focusing lens, for example, if the overlap of adjacent light beams is too large, the performance of high sensitivity and low crosstalk will not be improved regardless of the structure after the re-focusing lens. At the position of the optical path length g from the focusing lens, the optical axes of the paired focusing lenses and the re-focusing lens are aligned and the same number of re-focusing lenses as the focusing lenses are arranged at intervals p. By setting the effective diameter of the re-focusing lens to be equal to the effective diameter of the focusing lens, that is, to D, the loss of the amount of detection light performed by the re-focusing lens can be suppressed, and a re-focusing lens array can be constructed. The sensor is arranged at a position where the optical path length from the focusing lens is larger than g in the rear section of the re-focusing lens. If considered in this way, it can be seen that even when using a re-focusing lens, Figure 3 or Figure 4 The conditions for achieving high sensitivity and low crosstalk also remain unchanged.

[0138] Figure 8 1 is a schematic diagram showing an example of a multicolor detection device using a wavelength dispersion element and a re-focusing lens array. Figure 8 (a) is a schematic diagram of the multicolor detection device viewed from a direction perpendicular to a plane including the optical axes of the condenser lens 2, Figure 8 (b) is a cross-sectional schematic diagram showing a multi-color detection device including a condenser lens 2 and a pair of re-condenser lenses 33, and the optical axes thereof are perpendicular to the arrangement direction of the condenser lens array and the re-condenser lens array. Figure 8 (c) is a diagram showing a shadow 42 detected by the two-dimensional sensor 37. Here, an example of performing three-color detection is shown.

[0139] like Figure 8 As shown in (a), the light from the light emitting point 1 is focused by the focusing lens 2 to form a light beam 9, which is then transmitted through the long pass filter 10. Figure 6 Same. Then, Figure 8 As shown in (b), each light beam 9 is incident in parallel on a common transmission diffraction grating 31 as a wavelength dispersion element, and is wavelength dispersed in a direction perpendicular to the light-emitting point array direction, and is refocused by a refocusing lens 33, and then incident on a two-dimensional sensor 37. Here, light beams 34, 35, and 36 represent light beams of the center wavelengths of A, B, and C fluorescence, respectively. Figure 8 As shown in (c), a wavelength dispersion image 41 of light emitted from each light emitting point 1 is obtained on an image 42 of the two-dimensional sensor 37. Here, images 38, 39 and 40 in the wavelength dispersion image 41 represent images of the central wavelengths of the A, B and C fluorescence, respectively.

[0140] exist Figure 8 In the multi-color detection device shown, the diameter d of each light-emitting point 1, the interval p between each light-emitting point 1 and each focusing lens 2, the focal length f of each focusing lens 2, the effective diameter D, and the optical distance g between each focusing lens 2 and each re-focusing lens 33 satisfy any one of Formulas (3) to (7), (8) to (12), and (16) to (18), thereby achieving the predetermined high sensitivity and low crosstalk, and realizing the miniaturization and low cost of the detection device.

[0141] Figure 8 In FIG. 1 , it is shown that when a re-focusing lens is used, the optical distance between the focusing lens and the re-focusing lens is set to g, so as to satisfy any one of the equations (3) to (7), (8) to (12), and (16) to (18). Similarly, an optical element that causes each light beam to advance in different directions or an optical element that expands the interval between each light beam may be arranged at a position at an optical distance g from a focusing lens that individually focuses the light from each light-emitting point array into a light beam, so that the light beam satisfies any one of the equations (3) to (7), (8) to (12), and (16) to (18).

[0142] For example, Fig. 9 In, through Figure 6 A prism 43 is arranged as an optical element at the rear stage of the long pass filter 10 to make the light beams 9 advance in different directions from each other, and then make them enter the common two-dimensional color sensor 11 in parallel for detection. Fig. 9 (a) shows the multicolor detection device viewed from a direction perpendicular to a plane including the optical axes of the condenser lens 2, Fig. 9 (b) shows the image 12 detected by the two-dimensional color sensor 11. The prism 43 changes the direction and angle of refraction according to the different incident positions of the light beams 9, thereby widening the interval between the refracted light beams 44. If the optical distance between the focusing lens 2 and the prism 43 is set to g, equations (1) to (18) remain unchanged and become the conditions for high sensitivity and low crosstalk. In addition, Figure 6 Compared with the case of , the intervals of the light-emitting point images 45 on the image 12 of the two-dimensional color sensor 11 are enlarged, thereby avoiding the increase of crosstalk after the prism 43, and making it easier to independently detect the light from each light-emitting point 1. In addition, since the intervals of the light-emitting point images 45 are large, it is not necessary to Fig. 9 The sensor shown is a common two-dimensional color sensor, but each light beam 44 can also be detected by a separate color sensor. Figure 7In the case of the structure, it is preferred to arrange the prism 43 between the long pass filter 10 and the dichroic mirror array. Since the optical path length between the focusing lens 2 and the prism 43 can be smaller than the optical path length between the focusing lens 2 and the two-dimensional sensor 30, it is easier to meet the low crosstalk condition. Figure 7 In (c), the intervals of the light-emitting point images 25 to 28 in the light beam splitting direction do not change, but the intervals in the light-emitting point array direction are enlarged, so that crosstalk can be easily reduced.

[0143] As described above, by sharing the dichroic mirror, filter, total reflection mirror, diffraction grating, and sensor in multiple light-emitting points, the device structure can be simplified and installation can be facilitated. In addition, the overall size of the detection device becomes small. In addition, according to the above structure, the detection efficiency and spectral accuracy are the same in each light-emitting point, thereby reducing the deviation of sensitivity and color recognition.

[0144] Fig.10 is Figure 6 An example of an optical element is to configure an optical fiber 46 at the rear stage of the long pass filter 10. Each light beam 9 is input from the input end of the individual optical fiber 46 and emitted from the output end with the interval expanded, and the emitted light beams 47 are input in parallel to the common two-dimensional color sensor 11 for detection. Fig.10 (a) shows the multicolor detection device viewed from a direction perpendicular to a plane including the optical axes of the condenser lens 2, Fig.10 (b) shows the image 12 detected by the two-dimensional color sensor 11.

[0145] If the optical distance between the focusing lens 2 and the incident end of the optical fiber 46 is set to g, then equations (1) to (18) remain unchanged and become the conditions for high sensitivity and low crosstalk. Figure 6 Compared with the case of , the intervals of the light-emitting point images 48 on the image 12 of the two-dimensional color sensor 11 are enlarged, thereby avoiding the increase of crosstalk after the optical fiber 46, and making it easier to independently detect the light from each light-emitting point 1. In addition, since the intervals of the light-emitting point images 48 are large, it is not necessary to Fig.10 The sensor shown is a common two-dimensional color sensor, but individual color sensors can also be used to detect each light beam 47. Figure 7 In the case of a structure of Figure 7 The optical fiber 46 can be connected to the position of the two-dimensional sensor 30, that is, the rear section of the dichroic mirror array. Figure 7Thus, the intervals and arrangements of the luminous point images 25 to 28 can be arbitrarily set, thereby making it easy to detect each luminous point image by a separate sensor, or to detect each luminous point image by a desired one-dimensional sensor or two-dimensional sensor.

[0146] Hereinafter, examples of the present invention will be described.

[0147] [Example 1]

[0148] Fig.11 Schematic diagram showing an example of the device structure of a capillary array DNA sequencer. Fig.11 The analysis sequence is described. First, a plurality of capillaries 49 ( Fig.11 The sample injection end 50 of the four capillaries 49 (shown in the figure) is immersed in the cathode side buffer 60, and the sample elution end 51 is immersed in the anode side buffer 61 through the polymer block 55. The valve 57 of the pump block 55 is closed, and the internal polymer solution is pressurized by the syringe 56 connected to the pump block 55, so that the inside of each capillary 49 is filled with the polymer solution from the sample elution end 51 toward the sample injection end 50. Next, the valve 57 is opened, and different samples are injected into each capillary from the sample injection end 50, and then a high voltage is applied between the cathode 58 and the anode 59 by the power supply 62, thereby starting capillary electrophoresis. DNA marked by four colors of fluorescent substances is electrophoresed from the sample injection end 50 to the sample elution end 51.

[0149] The coating of each capillary 49 at the position (laser irradiation position 52) after electrophoresis for a certain distance from the sample injection end 50 is removed and each capillary 49 is arranged on the same plane. The laser beam 54 oscillated by the laser source 53 is focused and then introduced along the arrangement plane from the side, thereby irradiating the laser irradiation position 52 of each capillary 49 at the same time. DNA marked with four colors of fluorescent substances is electrophoresed inside each capillary 49 and excited when passing through the laser irradiation position 52, thereby emitting fluorescence. The light from the inside of each capillary 49 forms a light-emitting point array, and is formed by Figure 6 to Figure 10 The multicolor detection device represented in FIG. Fig.11 Each light emission is detected in a direction perpendicular to the paper surface).

[0150] In this embodiment, specifically Fig.11The case where a capillary DNA sequencer uses a multicolor detection device based on a dichroic mirror array is described. The laser irradiation positions 52 of four capillaries 49 with an outer diameter of 0.36 mm and an inner diameter of 0.05 mm are arranged on the same plane with an interval of p = 1 mm, and a laser beam 54 focused to a diameter of 0.05 mm is irradiated from the side of the arrangement plane, thereby obtaining a luminous point array with a number of M = 4 luminous points 1 and an effective diameter d = 0.05 mm arranged at an interval of p = 1 mm. Here, the effective diameter of the luminous point is set to be consistent with the inner diameter of the capillary. The wavelength of the laser beam 54 is 505 nm, and the four colors of fluorescence (luminescence maximum wavelength) are A fluorescence (540 nm), B fluorescence (570 nm), C fluorescence (600 nm), and D fluorescence (630 nm). The full width of the luminous point array is set to AW = p × (M-1) = 3 mm. Each condenser lens 2 of the condenser lens array 8, in which four condenser lenses 2 with a focal length of f = 1.5 mm and an effective diameter of D = 1 mm are arranged at intervals of p = 1 mm, condenses the light from each light-emitting point 1. The above d, p, f and D are basically equal in each light-emitting point and each condenser lens, but they are not necessarily equal. In this case, d, p, f and D are average values ​​of multiple light-emitting points and condenser lenses.

[0151] Fig.12 is a cross section of a multicolor detection device including the optical axis of a condenser lens 2 and perpendicular to the arrangement direction of the condenser lens array 8, but Figure 7 Differently, the optical axis of each condensing lens 2 is perpendicular to the advancing direction of the light beam generated by the dichroic mirror array after being split, that is, the optical axis of each condensing lens 2 is parallel to the sensor surface of the two-dimensional sensor 30. Figure 7 .in, Fig.12 The illustration of the light emitting point 1 is omitted.

[0152] A long pass filter 10 with a width of α = 5 mm, a thickness of β = 1 mm, and a depth of γ = 5 mm is arranged at a distance of 3 mm from the condenser lens 2 so that the optical axis of the condenser lens 2 is parallel to the normal line. Furthermore, dichroic mirrors 17, 18, 19, and 20 formed with a multilayer film or a single layer film are arranged at intervals of 5 mm on the lower right front side of a quartz substrate (refractive index n0 = 1.46) with a width of α = 5 mm, a thickness of β = 1 mm, and a depth of γ = 5 mm so that the normal line is inclined at 45° with respect to the optical axis of the condenser lens 2. Figure 7 Similarly, the dichroic mirror 20 can also be replaced by a total reflection mirror. Fig.12In the figure, the long-pass filter 10 and the dichroic mirrors 17 to 20 all show the side of α×β, and γ is the direction perpendicular to the paper surface. In addition, an anti-reflection film for reducing reflection loss is formed on the upper left front of the dichroic mirrors 17, 18, and 19. In addition, a light-shielding film for preventing the transmission of light is formed on the entire side of the dichroic mirrors 17, 18, 19, and 20 to prevent unexpected stray light. The right end of the dichroic mirror 17 is arranged at a position 5 mm away from the focusing lens 2 (i.e., a position 2 mm away from the long-pass filter 10). The upper and lower ends of the dichroic mirrors 17, 18, 19, and 20 are arranged on the same plane. A two-dimensional sensor 30 is arranged at a position 5 mm away from the lower ends of the dichroic mirrors 17, 18, 19, and 20. The above optical system elements are all arranged in the air. The air arrangement is to improve the light separation performance of the dichroic mirrors.

[0153] Fig.12 The 11 light beam elements 65 shown can divide the parallel light beam incident through the above-mentioned dichroic mirror array into four parts as designed, and show the light beam when the width of the light beam is the largest, and show the result of calculating the above-mentioned light path using the law of reflection and the law of refraction. Hereinafter, the above-mentioned maximum light beam width is referred to as the opening width 63 of the dichroic mirror array, and its size is represented by W. The opening width refers to the maximum width of the light beam that can well divide the light beam incident by the dichroic mirror array as designed. Generally, the opening width W is different from the width d'(s) of the light beam formed by the condenser lens 2 of the light path length s from the condenser lens 2, which is formed by the light from the light point 1. d'(s) is the actual width of the light beam, and in contrast, W represents the maximum value of the width of the light beam that the multi-color detection device can accept under the given conditions. That is, in order to split the light beam without causing loss of the light amount of the light beam after being condensed by the condenser lens 2, it is preferably set to W≥d'(s). The larger W is, the greater the tolerance for the deviation between the central axis of the light beam focused by the focusing lens 2 and the central axis of the opening width 63 is, and thus it is preferred. The 11 light beam elements 65 advancing leftward from the focusing lens 2 are set to be equally spaced and parallel to each other within the opening width 63 of the light beam.

[0154] like Fig.12 As shown in FIG. 1 , each light beam element 65 moves in parallel to the upper side in sequence due to internal refraction when passing through each dichroic mirror 17 to 19. In order to reduce this effect, the optical axis of the condenser lens 2 and the center of the light beam element emitted from the condenser lens are arranged to be lower than the center of the lower right front side of the dichroic mirror 17. Fig.12It is clear that the optical distance between the condenser lens 2 and the two-dimensional sensor 30, that is, the optical path length s, is different for each of the four divided light beams, and the light beam that passes through the dichroic mirrors 17 to 19 and is reflected by the dichroic mirror 20 has the largest optical path length 64. Hereinafter, when there are multiple optical paths between the condenser lens and the sensor, the optical path length 64 of the longest optical path is used as the optical path length of the light detection device, and its size is represented by g. Fig.12 The multi-color detection device is calculated to have an opening width 63 of W = 2.1 mm and an optical path length 64 of g = 29 mm. The above W and g are basically equal in each light-emitting point and each focusing lens, but they are not necessarily equal. In this case, W and g are the average values ​​of multiple light-emitting points and focusing lenses.

[0155] By setting the optical distance between the light-emitting point 1 and the focusing lens 2 with a focal length of f=1.5mm to about 1.58mm, the light from the light-emitting point 1 is formed at a position with an optical distance of g=29mm from the focusing lens 2, with an image magnification m=18.3 obtained from equation (1) and a diameter d'=0.92mm of the light-emitting point image 7 of the light-emitting point 1 with a diameter d=0.05mm obtained from equation (2). Since the effective diameter D=1mm of the focusing lens is larger than the above d', the optical path length s from the focusing lens 2 is 0mm≤s≤29mm, and in contrast, d'(s)≤1mm, and thus d'(s)≤W=2.1mm. Therefore, the light beams divided into four parts by the dichroic mirrors 17 to 20 can all reach the two-dimensional sensor 30 without loss.

[0156] In the above multi-color detection device, it can be seen that equations (4), (9) and (18) are satisfied, thereby obtaining high sensitivity and low crosstalk conditions of a relative detection light quantity of more than 100%, a strict relative detection light quantity of more than 100% and a crosstalk of 0%. The size of the multi-color detection device is the full width AW of the light-emitting point array = 3 mm, as shown in FIG. Fig.12 As shown, the volume of a rectangular parallelepiped (668 mm) defined by a width of 24.2 mm in the optical axis direction of the condenser lens 2 and a width of 9.2 mm in a direction perpendicular to the optical axis of the condenser lens 2 and the light-emitting point array can be reduced. 2 ) is small. That is, compared with the case of Patent Document 1, the overall size of the fluorescence detection device can be miniaturized to 1 / 2400 times. In addition, the optical elements used are all tiny, which can greatly reduce the cost. The above m and d' are basically equal in each light-emitting point and each focusing lens, but they are not necessarily equal. In this case, m and d' are average values ​​of multiple light-emitting points and focusing lenses.

[0157] [Example 2]

[0158] In Example 1 Fig.12In the multicolor detection device, the arrangement directions of the dichroic mirror array, the condenser lens array, and the light-emitting point array are arranged parallel to the sensor surface of the two-dimensional sensor 30, so that no three-dimensional obstruction occurs between them. However, since the arrangement plane of each capillary 49 of the capillary array is perpendicular to the sensor surface of the two-dimensional sensor 30, there is a situation where three-dimensional obstruction occurs between them, which becomes a problem in the device structure. Therefore, in this embodiment, a multicolor detection device that solves this problem is proposed.

[0159] The structure of the capillary DNA sequencer from the capillary array to the focusing lens array is the same as that of Example 1, except that Fig.12 The structure is replaced by Fig.13 structure. Fig.13 is a cross-sectional schematic diagram of a multicolor detection device including an optical axis of a condenser lens 2 and perpendicular to the arrangement direction of the condenser lens array 8, and Figure 7 Similarly, the optical axis of each condenser lens 2 is parallel to the advancing direction of the divided light beam, that is, the optical axis of each condenser lens 2 is perpendicular to the sensor surface of the two-dimensional sensor 30. A long pass filter 10 having a width α=5 mm, a thickness β=1 mm, and a depth γ=5 mm is arranged at a distance of 3 mm from the condenser lens 2 so that the optical axis of the condenser lens 2 is parallel to the normal line. In addition, a dichroic mirror 17 having a multilayer film is arranged at intervals of 5 mm on the upper left front side of a quartz substrate (refractive index n0=1.46) having a width α=5 mm, a thickness β=1 mm, and a depth γ=5 mm so that the normal line is inclined at 45° with respect to the optical axis of the condenser lens 2, and dichroic mirrors 18, 19, and 20 having a multilayer film or a single layer film are arranged on the lower right front side. An anti-reflection film for reducing reflection loss is formed on the lower right front side of the dichroic mirror 17 and the upper left front sides of the dichroic mirrors 18 and 19. Furthermore, light-shielding films for preventing transmission of light are formed on the entire side surfaces of the dichroic mirrors 17 , 18 , 19 , and 20 , thereby preventing unexpected stray light.

[0160] The upper end of the dichroic mirror 17 is arranged at a distance of 5 mm from the condenser lens 2. The upper and lower ends of the dichroic mirrors 17, 18, 19, and 20 are arranged on the same plane. A two-dimensional sensor 30 is arranged at a distance of 5 mm from the lower ends of the dichroic mirrors 17, 18, 19, and 20. The above optical system elements are arranged in the air. Fig.12 Similarly, 11 beam elements 65 are shown. As a result, for Fig.13 The multi-color detection device is used to calculate the opening width 63 as W = 1.7 mm, the optical path length 64 as g = 28 mm, and the same Fig.12 Equal performance.

[0161] By setting the optical distance between the light-emitting point 1 and the focusing lens 2 to about 1.58 mm, the light from the light-emitting point 1 is imaged at an image magnification of m=17.7 and a diameter d'=0.88 mm of the light-emitting point image 7 at an optical distance g=28 mm from the focusing lens 2. Since the effective diameter D=1 mm of the focusing lens is larger than the above d', the optical distance s from the focusing lens is 0 mm≤s≤28 mm, and in contrast, d'(s)≤1 mm, and thus d'(s)≤W=1.7 mm. Therefore, the light beams divided into four parts by the dichroic mirrors 17 to 20 can all reach the two-dimensional sensor 30 without loss.

[0162] In the above multi-color detection device, it can be seen that equations (4), (9) and (18) are satisfied, thereby obtaining high sensitivity and low crosstalk conditions of a relative detection light quantity of more than 100%, a strict relative detection light quantity of more than 100% and a crosstalk of 0%. The size of the multi-color detection device is the full width AW of the light-emitting point array = 3 mm, as shown in FIG. Fig.13 As shown, the volume of the rectangular parallelepiped (818mm) defined by the width 14.2mm in the optical axis direction of the condenser lens 2 and the width 19.2mm in the direction perpendicular to the optical axis of the condenser lens 2 and the arrangement direction of the light-emitting point array 2 ) is small. That is, compared with the case of Patent Document 1, the overall size of the fluorescence detection device can be miniaturized to 1 / 2000 times. In addition, the optical elements used are all tiny, which can significantly reduce costs.

[0163] exist Fig.13 In the multi-color detection device, the arrangement directions of the color separation mirror array, the focusing lens array, the light-emitting point array and the arrangement plane of the capillary array are all arranged to be parallel to the sensor surface of the two-dimensional sensor 30, so as to avoid three-dimensional obstruction and thus facilitate their installation.

[0164] [Example 3]

[0165] The conditions of Example 1 and Example 2 were studied in which the inner diameter of each capillary 49, that is, the diameter of the light emitting point 1, which was d=0.05 mm, was enlarged to d=0.075 mm. Fig.13 The multi-color detection device, at the position of optical path length g = 28mm, makes the light from the light point 1 imaged with image magnification m = 17.7 and diameter d' = 1.33mm of the light point image 7. Since the above d' is larger than the effective diameter D = 1mm of the focusing lens, the optical distance s from the focusing lens is 0mm≤s≤28mm, and d'(s)≤1.33mm, so d'(s)≤W=1.7mm holds. Therefore, Fig.13 Similarly, the light beams divided into four parts by the dichroic mirrors 17 to 20 can all reach the two-dimensional sensor 30 without any loss.

[0166] However, in the above multicolor detection device, Fig.13 Similarly, equations (4) and (9) are satisfied, but equation (18) is not satisfied, while equation (17) is satisfied. Therefore, it can be seen that the relative detection light quantity is 100% or more, and the strict relative detection light quantity is 100% or more and the crosstalk is 25% or less, which is a high sensitivity and low crosstalk condition. Fig.13 Therefore, in this embodiment, the image magnification m and the diameter d' of the light-emitting point image 7 are reduced by reducing the optical path length g, thereby reducing crosstalk. In order to reduce the optical path length g, it is considered that the size and arrangement interval of each dichroic mirror 17 to 20 can be reduced.

[0167] Fig.14 Show that Fig.13 The result is that the long-pass filter 10 and the dichroic mirrors 17 to 20 under the conditions are reduced from width α=5mm, thickness β=1mm, and depth γ=5mm to width α=2.5mm, thickness β=1mm, and depth γ=5mm, and the arrangement interval of the dichroic mirrors 17 to 20 is reduced from x=5mm to x=2.5mm.

[0168] at this time, Fig.14 , the left end of the dichroic mirror 17 and the right end of the dichroic mirror 18 are aligned in the lateral direction (direction perpendicular to the optical axis of the condenser lens 2). Similarly, the left end of the dichroic mirror 18 and the right end of the dichroic mirror 19, and the left end of the dichroic mirror 19 and the right end of the dichroic mirror 20 are aligned in the lateral direction, respectively. Other conditions are the same as Fig.13 Equal. Fig.14 With Fig.13 The same scale is used to show the

[0169] As a result, the optical path length 64 can be reduced from g = 28 mm to g = 19 mm. Therefore, the image magnification m = 11.7 is obtained from equation (1), and the diameter of the light-emitting point image d' = 0.88 mm is obtained from equation (2). In addition to equations (4) and (9), equation (18) is also satisfied. In other words, the relative detection light quantity is 100% or more, and the strict relative detection light quantity is 100% or more and the crosstalk is 0% or less. High sensitivity and low crosstalk conditions are achieved. However, it can be seen that the opening width 63 is greatly reduced from W = 1.7 mm to W = 0.03 mm, that is, the light beam condensed by the condenser lens 2 cannot reach the two-dimensional sensor 30 without loss. This is because, as the size and interval of each dichroic mirror are reduced, the relative ratio relative to the thickness β = 1 mm increases, so that when the light beam passes through each dichroic mirror, the influence of the sequential upward parallel movement due to internal refraction cannot be ignored.

[0170] Therefore, in order to eliminate the above effects, Fig.15As shown in FIG. 1 , the dichroic mirrors 17 to 20 are changed from being arranged in the same plane to being arranged with a difference. That is, according to the above parallel movement, the relative positions of the dichroic mirrors 17 to 20 in the optical axis direction of the condenser lens 2 are changed from being in the same plane. Fig.14 First, the upper end of the dichroic mirror 17 is arranged at a distance of 6.3 mm from the focusing lens 2. Next, compared with the lower end of the dichroic mirror 17, the lower end of the dichroic mirror 18 is directed to the side opposite to the advancing direction of the light beam after passing through the dichroic mirror 17, that is, Fig.15 Next, compared with the lower end of the dichroic mirror 18, the lower end of the dichroic mirror 19 is opposite to the direction of travel of the light beam reflected by the dichroic mirror 18, that is, Fig.15 Finally, compared with the lower end of the dichroic mirror 19, the lower end of the dichroic mirror 20 is directed to the side opposite to the advancing direction of the light beam reflected by the dichroic mirror 19, that is, Fig.15 The upper side deviation in z=0.3mm.

[0171] Based on the above results, it is clear that the opening width 63 can be changed from Fig.14 The W=0.03mm of the optical path is greatly expanded to W=1.3mm. On the other hand, the optical path length 64 is g=21mm, which is Fig.14 The situation is slightly increased, so the image magnification m = 13 is obtained from formula (1), and the diameter of the light-emitting point image is d' = 0.98mm from formula (2). Since the effective diameter D = 1mm of the focusing lens is larger than the above d', the optical path length s from the focusing lens is 0mm≤s≤21mm, and in contrast, d'(s)≤1mm, so d'(s)≤W=1.3mm holds. Therefore, the light beams divided into four parts by the dichroic mirrors 17 to 20 can all reach the two-dimensional sensor 30 without loss.

[0172] In the above multicolor detection device, Fig.14 The same, it can be known that equations (4), (9) and (18) are satisfied, thereby obtaining the high sensitivity and low crosstalk conditions of a relative detection light quantity of more than 100%, a strict relative detection light quantity of more than 100% and a crosstalk of 0%. The size of the multi-color detection device is the full width AW of the light-emitting point array = 3 mm, such as Fig.15 As shown, the volume of a rectangular parallelepiped (414 mm) defined by a width of 13.8 mm in the optical axis direction of the condenser lens 2 and a width of 10 mm in a direction perpendicular to the optical axis of the condenser lens 2 and the arrangement direction of the light-emitting point array can be reduced. 2 ) is small. That is, compared with the case of Patent Document 1, the overall size of the fluorescence detection device can be miniaturized to 1 / 3900 times. In addition, the optical elements used are all tiny, which can significantly reduce costs.

[0173] [Example 4]

[0174] It is clear from the above embodiments that in a multi-color detection device using a dichroic mirror array, it is important to achieve both the expansion of the opening width W and the reduction of the optical path length g, which are in a compromise relationship. This becomes difficult as the device becomes smaller. The adjustment of the arrangement spacing x, the height difference y and z of the dichroic mirror array shown in Example 3 is an effective solution to solve this problem. In this embodiment, the above-mentioned arrangement spacing and height difference configuration are generalized to derive a general solution that achieves both the expansion of the opening width W and the reduction of the optical path length g. The end of the configuration of the dichroic mirror of the present invention is characterized in that the thickness β of the dichroic mirror is taken into account. In the past, since the size of the dichroic mirror (α and γ) was sufficiently large, it would not be a problem even if β was not considered, but when they become small, it is important to consider the configuration of β. In particular, when each dichroic mirror is not set in the air but is set inside a glass material, it is considered that β=0 is not a problem.

[0175] Fig.16 It is a cross-sectional schematic diagram of a multi-color detection device including an optical axis of a condenser lens 2 and perpendicular to the arrangement direction of the condenser lens array 8. When the width α and thickness β of the dichroic mirror are set, the structure of the multi-color detection device is shown in which the opening width W is set to the maximum and the optical path length g is set to the minimum. Fig.15 FIG. 2 is a diagram of a condenser lens 2, a long pass filter 10, and a two-dimensional sensor 30.

[0176] Fig.16 In the example, the light beam 70 with an opening width W incident from top to bottom is repeatedly reflected and transmitted by the dichroic mirrors M(1), M(2), M(3), ..., and M(N) in sequence, thereby obtaining the light beams F(1), F(2), F(3), ..., and light beam F(N) emitted from top to bottom. The number of dichroic mirrors, i.e., the number of divisions of the light beam 70, is Fig.16 As an example, four are provided, but in this embodiment, N (N ≥ 2) are generally provided. Of course, the Nth color separation mirror M(N) can also be replaced by a total reflection mirror. Figure 12 to Figure 15 As shown in FIG. 1 , the optical path length g of the luminescence detection device is defined by the optical path length of the maximum length of the optical path from the focusing lens 2 to the sensor 30. In the following, as a part of the optical path length g, Fig.16 As shown, the optical path length L of the dichroic mirror array is defined by the optical path length from a position at the same height as the uppermost end of the dichroic mirror array on the optical axis of the light beam 70 (the upper end of the dichroic mirror M(N)) to a position at the same height as the lowermost end of the dichroic mirror array on the optical axis of the light beam F(N) (the lower end of the dichroic mirror M(1)).

[0177] Each dichroic mirror has an optical film formed on at least one front surface of a transparent substrate having a refractive index n0, and is arranged in air at intervals x. For each dichroic mirror, define Fig.16 The normal vector in the direction from the upper left front to the upper left. Each dichroic mirror is used to make each normal vector relative to the direction opposite to the forward direction of the light beam 70 ( Fig.16 The surface is tilted at an angle θ0 (0≤θ0≤90°) in a direction from bottom to top. Fig.16 In the figure, θ0=45° is used for depiction, but in the present embodiment, an arbitrary θ0 of 0°≤θ0≤90° is used. In summary, the dichroic mirrors M(1) to M(N) are arranged substantially parallel to each other and substantially at equal intervals. Furthermore, the lower end of the dichroic mirror M(2) is arranged so as to deviate upward by y relative to the lower end of the dichroic mirror M(1), that is, deviate by y in the direction opposite to the advancing direction of the light beam F(1). In addition, the lower end of the dichroic mirror M(3) is arranged so as to deviate upward by z relative to the lower end of the dichroic mirror M(2), that is, deviate by z in the direction opposite to the advancing direction of the light beam F(2). Similarly, assuming that 3≤n≤N, the lower end of the dichroic mirror M(n) is arranged so as to deviate upward by z relative to the lower end of the dichroic mirror M(n-1), that is, deviate by z in the direction opposite to the advancing direction of the light beam F(n-1).

[0178] At this time, the incident angle of the light beam on the incident surface of the dichroic mirror M(1) is θ0, and the refraction angle θ1 of the light beam on the incident surface (upper left front side) of the dichroic mirror M(1) is as follows.

[0179] [Formula 19]

[0180] θ1=sin -1 (1 / n0×sinθ0)

[0181] Furthermore, the incident angle of the light beam on the incident surface (lower right front) of the dichroic mirrors M(2) to M(N) is 90°-θ0, and the refraction angle θ2 of the light beam on the incident surface of the dichroic mirrors M(2) to M(N) is as follows.

[0182] [Formula 20]

[0183] θ2=sin -1 (1 / n0×sin(90°-θ0))

[0184] In the light beam 70, the right end is indicated by a dotted line as the light beam right end 66, and the left end is indicated by a dashed line as the light beam left end 67, and the diagram is traced to the right end and left end of each light beam F(1), F(2), F(3), ..., and light beam F(N).

[0185] according to Fig.16, the following are two conditions for the optimal mode that sets the opening width W to the maximum and the optical path length L to the minimum. First, the right end 66 of the light beam passes through or passes by the corner 69 at the left end of the dichroic mirrors M(1), M(2), ..., and M(N-1) shown in △. Second, the left end 67 of the light beam passes through or passes by the corner 68 at the lower end of the dichroic mirror M(1) shown in ○ and the corner 69 at the left end of the dichroic mirrors M(2), ..., and M(N-1). According to the above conditions, by Fig.16 First, the interval x between the dichroic mirrors M(1) to M(N) is expressed as follows in the best mode.

[0186] [Formula 21]

[0187] x=x0=cosθ0×α+sinθ0×β

[0188] And, the opening width W is expressed as follows in the best mode.

[0189] [Formula 22]

[0190] W=W0=a W ×α+b W ×β

[0191] Here, it is expressed as follows.

[0192] [Formula 23]

[0193] a W ≡cosθ0

[0194] [Equation 24]

[0195] b W ≡-cosθ0×tanθ1

[0196] In addition, the optical path length L is expressed as follows in the best mode.

[0197] [Formula 25]

[0198] L=L0=a L ×α+b L ×β

[0199] Here, it is expressed as follows.

[0200] [Equation 26]

[0201] a L ≡(N-1)×cosθ0+sinθ0

[0202] [Formula 27]

[0203] b L≡(N-2) / cosθ0×(2×sin(90°-θ0-θ2)+1-sin(θ0+θ2))+(N-2)×sinθ0+2×cosθ0

[0204] On the other hand, the step differences y and z of the dichroic mirrors M(1) to M(N) are expressed as follows in the best mode.

[0205] [Equation 28]

[0206] y=y0=cosθ0×β

[0207] [Formula 29]

[0208] z=z0=sin(90°-θ0-θ2) / cosθ2×β

[0209] As described above, x0, W0, L0, y0, and z0 are all associated with α and β.

[0210] The above α, β, n0, θ0, x and z are basically the same in each dichroic mirror and the total reflection mirror, but they are not necessarily the same. In this case, α, β, n0, θ0, x and z are the average values ​​of the multiple dichroic mirrors.

[0211] By reversing the above equation, the minimum value W for obtaining the target opening width can be derived: min α, β and x. According to W0≥W min and formula (22) are expressed as follows (the equal sign is the best mode),

[0212] [Formula 30]

[0213] α≥-b W / a W ×β+1 / a W ×W min

[0214] And according to formula (21), it is expressed as follows (the equal sign is the best mode).

[0215] [Equation 31]

[0216] x≥(sinθ0-b W / a W ×cosθ0)×β+1 / a W ×cosθ0×W min

[0217] Similarly, the maximum value L for obtaining the target optical path length can be derived: max α, β and x. According to L0≤L max and formula (25) are expressed as follows (the equal sign is the best mode),

[0218] [Formula 32]

[0219] α≤-b L / a L ×β+1 / a L ×L max

[0220] And according to formula (21), it is expressed as follows (the equal sign is the best mode).

[0221] [Formula 33]

[0222] x≤(sinθ0-b L / a L ×cosθ0)×β+1 / a L ×cosθ0×L max

[0223] Above W min and L max Basically, W is equal in each light-emitting point and each focusing lens, but not necessarily. min and L max It is the average value among multiple light-emitting points and focusing lenses.

[0224] Fig.17 As an example, when N = 4, n0 = 1.46, θ0 = 45°, the horizontal axis β and the vertical axis x show the range that satisfies equations (31) and (33). As a parameter, W min = 0.5, 1, 2, 3 and 4 mm, and L max = 5, 10, 20, 30 and 40 mm, ↑ represents the range above the straight line, ↓ represents the range below the straight line. min =0.5mm and L max =20mm, it can be seen that the selected Fig.17 Middle↑W min = 0.5 and ↓L max = 20.

[0225] On the other hand, with respect to the given diameter d of the light-emitting point and the interval p of the light-emitting point array, the focal length f of the focusing lens and the optical path length g between the focusing lens and the sensor are selected to satisfy the high sensitivity condition of any one of equations (3) to (7) or (8) to (12), and are set to W min =d',L max=g, if equations (31) and (33) are satisfied, a small multi-color detection device using a dichroic mirror array with high sensitivity conditions can be constructed. Here, according to equation (2), d' is set to (g-f) / f×d. Similarly, with respect to the given diameter d of the light-emitting point and the spacing p of the light-emitting point array, the focal length f of the focusing lens that satisfies any low crosstalk condition in equations (16) to (18) and the optical path length g between the focusing lens and the sensor are selected and set to W min =d',L max =g, if equations (31) and (33) are satisfied, a small multi-color detection device using a dichroic mirror array with low crosstalk conditions can be constructed. Of course, the focal length f of the condenser lens and the optical path length g between the condenser lens and the sensor are selected to satisfy both the high sensitivity condition of any one of equations (3) to (7) or equations (8) to (12) and the low crosstalk condition of any one of equations (16) to (18), and are set to W min =d',L max =g, if equations (31) and (33) are satisfied, a small multi-color detection device using a dichroic mirror array with high sensitivity and low crosstalk conditions can be constructed.

[0226] For example, according to Example 1, when N=4, n0=1.46, θ0=45°, β=1mm, x=5mm relative to d=0.05mm, p=1mm, f=1.5mm, D=1mm, g=29mm, all of Formulas (4), (9), (18), (31) and (33) are satisfied, and it can be seen that a small multi-color detection device using a high-sensitivity and low-crosstalk color separation mirror is obtained.

[0227] Next, the interval x of each dichroic mirror M(1) to M(N) will be studied in detail. As described above, in the optimum mode, x0 in equation (21) is the best. The following will examine in detail how much the effect can be obtained by deviating from the optimum mode. Fig.18 The solid line shown is the calculated interval x and Fig.15 The result is the relationship between the opening width W obtained by the dichroic mirrors 17 and 18. Generally, as the total number N of dichroic mirrors increases, the total opening width may be smaller than the above result. Here, the case of N=2 is used as an indicator for evaluation. Fig.15 The condition x=x0=2.5mm is calculated by equation (21) when θ0=45° and β=1mm. However, Fig.18 As shown, the opening width W = 1.3mm becomes the maximum. When x < x0, W decreases in proportion to |x-x0|, x = 1.6mm and W = 0mm. On the other hand, when x > x0, W = 1.3mm and remains constant. On the other hand, Fig.18 The dashed line shows the interval x and Fig.15Here, when x=x0=2.5mm, ΔL=0mm is set, and it is displayed at the same height as W=1.3mm. In addition, the scales of the vertical axis of W (left side) and the vertical axis of ΔL (right side) are aligned, and the vertical axis of ΔL is reversed up and down. Generally, as the total number N of dichroic mirrors increases, ΔL may be larger than the above result. Here, the case of N=2 is used as an indicator for evaluation. ΔL increases in proportion to x, of course.

[0228] according to Fig.18 , the rate of increase of W relative to x when 1.6mm≤x≤2.5mm and the rate of increase of ΔL relative to x when 2.5mm≤x are both inclined and approximately equal to 1. In other words, it can be seen that the performance decreases in proportion to |x-x0|. In contrast, in the past, β was not considered, which was equivalent to β=0mm. At this time, assuming that the interval x0 in the case of the same configuration is x=1.8mm in formula (21), at this time from Fig.18 It can be seen that W = 0.4mm. In summary, in order to obtain the same or higher performance as before, it can be seen that 1.8mm≤x≤3.2mm is sufficient. Generally, Fig.16 In the equation ( ), 2≤n≤N is assumed, and by setting the arrangement interval x between the dichroic mirrors M(n) and M(n-1) as follows, the opening width W can be enlarged and the optical path length L can be shortened.

[0229] [Equation 34]

[0230] cosθ0×α≤x≤cosθ0×α+2×sinθ0×β

[0231] Next, the step differences y and z of the dichroic mirrors M(1) to M(N) are studied in depth. As described above, in the optimal mode, y0 and z0 of equations (28) and (29) are the best. The following is a detailed study of the extent to which the effect of the step difference configuration can be obtained by deviating from the optimal mode.

[0232] Fig.19 (a) is to calculate the order difference y and Fig.15 The result is the relationship between the opening width W obtained by the dichroic mirrors 17 and 18. Generally, as the total number N of dichroic mirrors increases, the total opening width W may be smaller than the above result. Here, the case of N=2 is used as an indicator for evaluation. Fig.15 The condition y=y0=0.7mm is calculated by equation (28) when θ0=45° and β=1mm. However, Fig.19 As shown in (a), the opening width W = 1.3 mm is the maximum. Moreover, W decreases in proportion to |y-y0|, and W = 0.6 mm when y = 0 mm and 1.4 mm, and W = 0 mm when y = -0.7 mm and 2.1 mm. Here, negative y indicates Fig.15Therefore, it can be seen that the effect of the step difference can be obtained by setting 0mm≤y≤1.4mm.

[0233] same, Fig.19 (b) is to calculate the order difference z and Fig.15 The result is the relationship between the opening width W obtained by the dichroic mirrors 18 and 19. Fig.15 The condition z=z0=0.3mm is calculated by equation (29) when θ0=45° and β=1mm. However, Fig.19 As shown in (b), the opening width W = 1.3 mm, which is the maximum. Moreover, W decreases in proportion to |z-z0|, and W = 1 mm when z = 0 mm and 0.6 mm, and W = 0 mm when y = -1.1 mm and 1.7 mm. Here, negative z indicates Fig.15 Therefore, it can be seen that the effect of the step difference can be obtained by setting 0mm≤z≤0.6mm. If the above content is generalized, it can be described as follows. Fig.16 In the embodiment, the end of the split light beam advancing side of the dichroic mirror M(2) is deviated by an angle y to the side opposite to the advancing direction of the split light beam relative to the end of the split light beam advancing side of the dichroic mirror M(1), and the opening width W can be enlarged and the optical path length L can be shortened by the following arrangement.

[0234] [Formula 35]

[0235] 0≤y≤2×cosθ0×β

[0236] Furthermore, 3≤n≤N is set, and the end of the split light beam on the advancing side of the dichroic mirror M(n) is deviated by z to the side opposite to the advancing direction of the split light beam relative to the end of the split light beam on the advancing side of the dichroic mirror M(n-1), and by the following setting, the opening width W can be expanded and the optical path length L can be shortened.

[0237] [Equation 36]

[0238] 0≤z≤2×sin(90°-θ0-θ2) / cosθ2×β

[0239] Above, such as Fig.15 as well as Fig.16 As shown in FIG. 1 , the structure in which the optical axis of the condenser lens is parallel to the advancing direction of the split light beam has been studied, but in the case of Fig.12 When they are perpendicular as shown, it is set to 2≤n≤N, so that the end of the split light beam on the advancing side of the dichroic mirror M(n) deviates from the end of the split light beam on the advancing side of the dichroic mirror M(n-1) to the side opposite to the advancing direction of the split light beam by z, and as shown in formula (36), the opening width W can be expanded and the optical path length L can be shortened.

[0240] [Example 5]

[0241] In the third and fourth embodiments, the dichroic mirror array has an enlarged opening width W and a reduced optical path length L by configuring the multiple dichroic mirrors with a step difference. In the present embodiment, the following scheme is proposed: when there is no step difference configuration, that is, when the multiple dichroic mirrors are configured on the same plane, more specifically, when the ends of the split light beams on the advancing direction of the dichroic mirrors are arranged side by side on the same plane, the enlarged opening width W and the reduced optical path length L are achieved.

[0242] Example 3 Fig.14 This is the result for the case where θ0 = 45°. Fig. 20 The results are shown for the case where θ0 = 50°. Fig.14 and Fig. 20 In any case, the dichroic mirrors 17 to 20 are arranged on the same plane. Fig.14 In the case of W = 0.03 mm, Fig. 20 can be greatly expanded to W = 0.9 mm. The maximum optical path length does not change in the two cases, L = 19 mm. Fig.14 Similarly, the image magnification m = 11.7, the diameter of the light-emitting point image d' = 0.88 mm, in addition to satisfying equations (4) and (9), it also satisfies equation (18). Therefore, it becomes a high sensitivity and low crosstalk condition with a relative detection light amount of 100% or more, a strict relative detection light amount of 100% or more and a crosstalk of 0% or less.

[0243] The reasons for achieving such an effect are discussed below. Fig.14 As shown in FIG. 1 , when θ0=45°, the light beam moves horizontally to the left in the space between different dichroic mirrors, and moves to the upper left inside each dichroic mirror, so that the light beam gradually moves upward each time it passes through the dichroic mirror, which limits the opening width W. In contrast, Fig. 20 As shown in FIG. 1 , by setting θ0=50°≥45°, the light beam advances toward the lower left direction in the space between different dichroic mirrors, and advances toward the upper left direction inside each dichroic mirror, so that the two cancel each other out, suppressing the vertical movement of the light beam each time it passes through the dichroic mirror, which causes the expansion of the opening width W. Therefore, θ0 is preferably greater than 45°, and there should be an optimal value that maximizes the opening width W.

[0244] Fig.21 is Fig.14 as well as Fig. 20The result of W after changing θ0 is calculated under the condition of θ0=45°. It can be seen that W increases from θ0=45°, reaches a maximum value of 0.92 mm at θ0=52°, and decays to almost zero at θ0=57°. In other words, it can be seen that W can be expanded by setting 45°≤θ0≤57°.

[0245] Next, let's generalize the above. Fig.16 The same discussion as Fig. 20 The following contents are derived from the geometric relationship. The refraction angle θ1 of the light beam at the incident surface of the dichroic mirror M(1) is shown in equation (19), and the refraction angle θ2 of the light beam at the incident surface of the dichroic mirrors M(2) to M(N-1) is shown in equation (20). The downward movement distance S↓ of the light beam moving toward the lower left in the space between different dichroic mirrors is solved by the following equation (37).

[0246] [Formula 37]

[0247] S↓=tan(2×θ0-90°)×tanθ0 / (tanθ0-tan(2×θ0-90°))×(x-β / cos(90°-θ0)

[0248] On the other hand, the upward movement distance S↑ of the light beam advancing toward the upper left direction inside each dichroic mirror is solved by the following formula (38).

[0249] [Equation 38]

[0250] S↑=1 / cosθ2×β×sin(90°-θ0-θ2)

[0251] Here, β represents the thickness of each dichroic mirror, and x represents the interval between each dichroic mirror. Fig. 20 As shown, in order to cancel out S↓ and S↑, it is preferable to set S↓=S↑. Therefore, θ0 in this optimal mode is set to θ0(BM).

[0252] When equations (37) and (38) are applied to Fig. 20 When the conditions are β = 1 mm and x = 2.5 mm, θ0(BM) = 50° is obtained. That is, Fig. 20 The structure of is the best model structure. However, according to Fig.21 , W becomes maximum when θ0=52°, which is 2° larger than the above θ0(BM). This shows that W can be slightly increased by making θ0 slightly larger than θ0(BM), that is, by making S↓ slightly larger than S↑ to gradually move the light beam to the lower left.

[0253] In summary, the conditions for intentionally increasing W compared to the conventional standard case of θ0=45° are as follows.

[0254] [Equation 39]

[0255] 45°≤θ0≤2×θ0(BM)-45°

[0256] Furthermore, if the above-mentioned deviation of 2° is taken into consideration, the more correct conditions are as follows.

[0257] [Formula 40]

[0258] 45°≤θ0≤2×θ0(BM)-43°

[0259] [Example 6]

[0260] Fig. 22 It is shown in Figure 6 Schematic diagram of a luminescence detection device in a case where the size of the luminous point is larger in the structure. The diameter of the luminous point 71 is d=0.5mm and the interval is p=1mm. Compared with Example 1, the size of the luminous point is one size larger. The luminous points 71 are each composed of a cubic reaction monomer of 0.5mm×0.5mm×0.5mm, and chemiluminescence is generated due to the internal chemical reaction. By investigating the temporal changes in the wavelength and intensity of the chemiluminescence for each luminous point 71, the sample after the introduction of each reaction monomer was analyzed. The focal length of the focusing lens 2 is f=1mm, the effective diameter is D=1mm, the interval is p=1mm, and the optical distance between the focusing lens 2 and the two-dimensional color sensor 11 is g=10mm. The arrangement direction of the luminous point 71 and the focusing lens 2 can also be not only in Fig. 22 (a) horizontally, in relation to Fig. 22 (a) is also arranged at equal intervals in the direction perpendicular to the paper surface. In this embodiment, no excitation light source is required, thus omitting Figure 6 The long pass filter 10.

[0261] like Fig. 22 As shown in (a), if the light-emitting point 71 is imaged on the sensor surface of the two-dimensional color sensor 11, then m = 9 is obtained from equation (1), and d' = 4.5 mm is obtained from equation (2). At this time, equations (6) and (10) are satisfied, so that the relative detection light quantity is 400% or more, and the strict relative detection light quantity is 200% or more. On the other hand, Fig. 22 As shown in the light beam 9 of (a), the crosstalk between different light-emitting points 71 is very large, and none of them satisfies equations (16) to (18).

[0262] Therefore, if Fig. 22 As shown in (b), a pinhole array 73 having pinholes 72 is arranged between each light emitting point 71 and the corresponding focusing lens 2. The diameter d0 of each pinhole 72 is d0≤d, and d0=0.1mm is set here. The interval between each pinhole is set to p=1mm. The arrangement direction of the pinholes can also be not only in Fig. 22 (b) horizontally, in relation to Fig. 22Here, not only the light-emitting point 71 but also the pinhole 72 is regarded as the light-emitting point. Fig. 22 As shown in (b), the pinhole 72 is imaged on the sensor surface of the two-dimensional color sensor 11 to form a pinhole image 74. The diameter of the pinhole image 74 is obtained from equation (2) as d'=0.9 mm. At this time, equation (18) is satisfied, and the crosstalk can be reduced to 0%. Fig. 22 (a) is the same, and equations (6) and (10) are satisfied, so that the relative detection light quantity is 400% or more, and the strict relative detection light quantity is 200% or more. However, this is based on the total light quantity passing through the pinhole 72, which is more Fig. 22 The relative detection light amount based on the total light amount emitted by the light emitting point 71 in (a) is small.

[0263] Fig.23 It shows the application and Fig. 22 (b) Schematic diagram of another example of the same detection device. Fig.23 As shown in (a), the diameter of the light-emitting point 75 is d = 0.01 mm, the interval p' = 0.1 mm, and the focal length of the condenser lens 2 is f = 1 mm, the effective diameter D = 1 mm, the interval p = 1 mm, and the optical path length g = 10 mm between the condenser lens 2 and the two-dimensional color sensor 11. In other words, unlike the above embodiment, the interval between the light-emitting points and the interval between the condenser lenses are different, which is an example of the case where p' < p. The light-emitting point 75 and the condenser lens 2 can also be Fig.23 If the light emitting points 75 are imaged on the sensor surface of the two-dimensional color sensor 11, the light from the comparison Fig. 22 In the case of (a), the light emitted from a plurality of light emitting points 75 in a wider range is condensed by each condenser lens 2, so that the crosstalk is further increased.

[0264] Therefore, if Fig.23 As shown in (b), a pinhole array 73 having pinholes 72 is arranged between each light emitting point 75 and the corresponding focusing lens 2. Each pinhole 72 is arranged to coincide with the position of the corresponding focusing lens 2. The diameter of each pinhole 72 is d0 = 0.1 mm, the interval p = 1 mm, and d0 ≥ d. The pinholes can also be arranged at the same position as the corresponding focusing lens 2. Fig.23 Here, by making the light-emitting point array and the pinhole array 73 sufficiently close to each other, each light-emitting point 75 and each pinhole 72 are imaged on the sensor surface of the two-dimensional color sensor 11, thereby forming a light-emitting point image 76 and a pinhole image 74 respectively. From equation (2), it can be concluded that the diameter of the pinhole image 74 is 0.9 mm, and the crosstalk between different pinhole images 74 is 0%, which is consistent with Fig. 22(b) is the same. On the other hand, each condensing lens 2 condenses the light emitted from the average of two light-emitting points 75 through each pinhole 72, thereby forming each light-emitting point image 76 inside each pinhole image 74 on the sensor surface of the two-dimensional color sensor 11. The diameter of each light-emitting point image 76 is 0.09 mm, and the interval is 0.9 mm, so there is no crosstalk between different light-emitting point images 76.

[0265] Fig.23 (b) The structure is capable of performing multi-color detection of light from light-emitting points arranged at intervals narrower than the intervals of the focusing lens with high sensitivity and low crosstalk, but only a part of the light-emitting points can be detected. Fig.23 In the case of (b), only an average of two luminous points out of 10 luminous points are detected. Fig.23 In (b), by shifting the relative positions of the light-emitting point array and the detection device after the pinhole array 73 in sequence, for example, in the direction of the arrow, that is, by scanning the light-emitting points to be detected among a plurality of light-emitting points, all light-emitting points can be detected.

[0266] In the above, a multicolor detection device using a two-dimensional color sensor is used, but of course, a detection device using a two-dimensional monochrome sensor instead of a two-dimensional color sensor may also be used. Figure 7 A multicolor detection device using a dichroic mirror array is shown.

[0267] Fig.24 (a) is a schematic diagram showing an example of a multicolor detection device using a dichroic mirror array used in this embodiment, Figure 7 (b) and Fig.23 (b) Structure. Fig.24 In (a), the light beams 21, 22, 23 and 24 have different optical path lengths. Therefore, if any one of the corresponding luminous point images 25, 26, 27 and 28 is imaged on the sensor surface of the two-dimensional sensor 30, the other luminous point images will not be imaged on the above-mentioned sensor surface, but will become a slightly blurred state. Figure 7 In the embodiment, a condenser lens 2 focuses a light point 75 to obtain four light point images 25, 26, 27 and 28, so that the above-mentioned blur will not become a cause of crosstalk. Fig.24 In (a), a focusing lens 2 focuses light on a plurality of light-emitting points 75, and each light-emitting point is divided into four light-emitting point images 25, 26, 27 and 28. As a result, due to the above-mentioned blur, the plurality of light-emitting point images 76 inside the pinhole image 74 overlap with each other, which becomes the cause of crosstalk.

[0268] Therefore, if Fig.24As shown in (b), in this embodiment, optical path length adjustment elements 77, 78 and 79 of different lengths are respectively inserted into the optical paths of the light beams 21, 22 and 23, so that the optical path lengths of the light beams 21, 22, 23 and 24 defined by the optical distance between the focusing lens 2 and the two-dimensional sensor 30 are adjusted to be approximately equal. The optical path length adjustment element is made of a transparent material with a refractive index greater than 1. For example, inside a material with a refractive index of 2, even if the distance is the same in space, the optical distance is twice that in the air. By having such a structure, the light-emitting point images 25, 26, 27 and 28 can be simultaneously imaged on the sensor surface of the two-dimensional sensor 30, and the above-mentioned blur and the resulting crosstalk can be avoided.

[0269] In the above, the detection object is an array of luminous points arranged at approximately equal intervals, but the detection object can be any one-dimensional, two-dimensional, or three-dimensional luminous distribution, and can also be used in conjunction with Fig.23 (b) The same luminescence detection device reconstructs the original luminescence distribution based on the detection results and forms an image. Fig.25 An example of such image formation is schematically shown.

[0270] like Fig.25 As shown in (a), the light emission distribution 80 is distributed two-dimensionally. Here, the light emission distribution 80 depicts the character "α". On the other hand, the pinhole array 73 is arranged parallel to and close to the light emission point distribution 80. Fig.25 (a) schematically shows the positional relationship and size relationship between the light-emitting point distribution 80 and the plurality of pinholes 72 included in the pinhole array 73. Here, 3×3=9 pinholes 72 are arranged at equal intervals in a two-dimensional manner. Since the light-emitting point distribution 80 is sufficiently close to each pinhole 72, Fig.25 In (a), the light from the portion of the light-emitting point distribution 80 that overlaps with each pinhole 72 passes through each pinhole 72 and is Fig.23 (b) Detection using the same luminescence detection device.

[0271] Fig.25 (b) shows the images of the nine pinholes 72 captured by the two-dimensional color sensor 11, namely, nine pinhole images 74, and the partial images of the light-emitting point distribution 80 detected through the nine pinholes 72, namely, nine light-emitting point distribution partial images 81. As in the embodiment so far, each pinhole 72 is enlarged and imaged. On the other hand, since the intervals between the pinholes 72 and the intervals between the pinhole images 74 are equal, Fig.25 As shown in (b), the gap between adjacent pinhole images 74 becomes narrower, but the mutual crosstalk is 0%. In addition, each luminous point distribution partial image 81 is confined inside the corresponding pinhole image 74, so the mutual crosstalk is also 0%.

[0272] Fig.25 (c) Show that Fig.25 (a) is an example in which the relative positions of the light-emitting point distribution 80 and the pinhole array 73 are shifted in the lateral direction. Fig.25 (a) shows that each pinhole 72 overlaps with a different portion of the light-emitting point distribution 80. Fig.25 As shown in (d), the light emitted from the overlapping portion is imaged and detected. As described above, by sliding the relative position of the light-emitting point distribution 80 and the pinhole array 73 in the horizontal and vertical directions in sequence and repeatedly photographing, it is possible to form an overall image of the light-emitting point distribution 80. At this time, if the relative position of the light-emitting point distribution 80 and the two-dimensional color sensor 11 is fixed, the overall image of the light-emitting point distribution 80 can be formed even without image processing.

[0273] [Example 7]

[0274] One of the problems in the installation of the present invention is how to align each light emitting point with each condenser lens with high precision and in a simple manner. This embodiment shows a method for achieving the alignment of a plurality of capillaries.

[0275] Fig.26 1 is a schematic cross-sectional view showing a structural example of a device 86 in which a plurality of capillaries 49 , a V-groove array in which the plurality of capillaries 49 are arranged, and a condenser lens array 8 are integrated. Fig.26 (a) shows a cross section perpendicular to the long axis of each capillary 49 at the irradiation position of the laser beam 54, Fig.26 (b) shows a cross section perpendicular to the long axis of each capillary 49 at a location other than the irradiation position of the laser beam 54, Fig.26 (c) shows a cross section including the major axis of any capillary. Fig.26 (a) Equivalent to Fig.26 (c) AA section, Fig.26 (b) equivalent to Fig.26 (c) BB cross section.

[0276] Fig.26 The device 86 shown includes a capillary array composed of a plurality of capillaries 49 and a sub-device 85. The sub-device 85 integrates a V-groove array device 84, which includes a V-groove array having a plurality of V-grooves 82 arranged at intervals p, and a condenser lens array device 83, which includes a condenser lens array 8 having a plurality of condenser lenses 2 arranged at intervals p. Fig.26In (a), each light-emitting point 1 is aligned with each V-groove 82 and the central axis of each condenser lens 2. By pushing the plurality of capillaries 49 to the V-groove 82, the plurality of capillaries 49 can be easily arranged on the same plane at a predetermined interval p. In addition, the structure of the sub-device 85 is preliminarily adjusted so that the irradiation position of the laser beam 54 of each capillary 49, that is, each light-emitting point 1 and each condenser lens 2 are at a desired distance. As a result, the luminescence from the light-emitting point 1 is condensed by the condenser lens 2 as desired.

[0277] like Fig.26 As shown in (a), in the cross section of the capillary 49 at the light emitting point 1, the focusing lens 2 of the sub-device 85 exists, but the V-groove 82 does not exist. Fig.26 As shown in (b), in the cross section of the capillary 49 on both sides of the light-emitting point 1, there is no focusing lens of the sub-device 83, but there is a V-groove 82. Fig.26 (c) shows a cross section of the capillary 49 in the long axis direction, and there is a condenser lens 2 in the center of the sub-device 85, and there are V-grooves 82 on both sides thereof. This realizes high-precision alignment of the capillary 49 using the V-grooves 82, and at the same time, it is ensured that the V-grooves 82 do not hinder the detection of light from the light-emitting point 1. If the above sub-device 85 is prepared in advance, it is possible to easily perform high-precision alignment of each light-emitting point 1 and each condenser lens 2 by simply pushing a plurality of capillaries 49 into each V-groove 82.

[0278] This embodiment can also be combined with any structure of the above embodiments. The sub-device 85 formed by integrating the V-groove array device 84 and the condenser lens array device 83 can be integrally formed by a processing method such as injection molding and imprinting, and can also be mass-produced at low cost. Of course, the sub-device 85 can also be completed by separately manufacturing the V-groove array device 84 and the condenser lens array device 83 and then combining them.

[0279] The sub-device may also be provided without a V-groove array. For example, the surface of the sub-device on the side where the capillaries are arranged may be formed into a plane without forming a V-groove array. Other mechanisms are required to adjust the arrangement interval of the plurality of capillaries, but by pushing each capillary to the above-mentioned plane of the sub-device, the distance between each capillary and each focusing lens, that is, the distance between each light-emitting point and each focusing lens, can be controlled. Alternatively, even if a V-groove is not provided, a structure for controlling the position of the capillary may be provided in the sub-device.

[0280] When the focal length of each condenser lens 2 in a direction parallel to the arrangement direction of the light-emitting point array is set to f1, and the focal length in a perpendicular direction is set to f2, in the above embodiment, it is set to f=f1=f2, but it can also be set to f1≠f2. For example, as in this embodiment, it is effective when the light-emitting point 1 exists inside the capillary 49. Since the capillary 49 is cylindrical, it has a lens effect in the arrangement direction of the light-emitting point array, but does not have a lens effect in the long axis direction of each capillary 49. Therefore, in order to efficiently condense the light from the light-emitting point 1 using the condenser lens 2, it is effective to cancel the difference caused by the direction of the lens effect of the capillary 49 mentioned above, and for this purpose, it is set to f1≠f2, specifically, f1<f2. This can be easily achieved by forming the surface of each condenser lens 2 into an aspherical shape. In addition, by making each condenser lens 2 a Fresnel lens, the thickness of the lens can be reduced, and the fluorescence detection device can also be made more compact. Of course, the use of the Fresnel lens is also effective when f1=f2.

[0281] Fig. 27 Shown in Fig.26 The pinhole array 73 is added between the light-emitting point array and the focusing lens array 8 in the embodiment. More specifically, the pinhole array 73 is sandwiched between the V-groove array device 84 and the focusing lens array device 83, and the whole device 86 is used as a sub-device. The amount of light that the focusing lens 2 focuses on the light emitted from the light-emitting point 71 varies depending on the light intensity. Fig. 22 (b) is limited by the presence of the pinhole 72. Fig. 27 As shown in (a), the diameter d0 of each pinhole 72 is larger than the diameter d of the light emitting point 1 (d0 ≥ d) so that the amount of light condensed by the condensing lens 2 from the light emitting point 1 will not be affected by Fig. 27 The presence of the pinhole 72 shown is limited. Fig. 22 The pinhole 72 in (b) actually effectively reduces the diameter of the light emitting point 71. Fig. 27 The pinhole 72 prevents the focusing lens 2 from focusing light other than the light emitted from the corresponding light emitting point 1. For example, the scattered light of the laser beam 54 on the outer surface of each capillary 49 generated when the laser beam 54 is irradiated to the capillary array is focused by the focusing lens 2, and its arrival at the sensor can be avoided or reduced. Alternatively, the light emitted from the adjacent light emitting point 1 is focused by the focusing lens 2, and its arrival at the sensor can be avoided or reduced. In summary, the light emitted from the light emitting point 1 can be detected with high sensitivity.

[0282] In order to prevent the above-mentioned unnecessary light from reaching the sensor, it is also effective to arrange a colored glass filter at an arbitrary position between the light-emitting point and the sensor. The colored glass filter may be used simultaneously with the above-mentioned pinhole, or only one of them may be used. The light beam formed by the above-mentioned unnecessary light being focused by the focusing lens 2 advances obliquely relative to the optical axis of the focusing lens 2 (that is, the optical axis of the light beam formed by the light from the light-emitting point 1 being focused by the focusing lens 2), and is thus difficult to be blocked by the above-mentioned long-pass filter and dichroic mirror (this is because they are designed for the light beam formed by the light from the light-emitting point 1 being focused by the focusing lens 2). In contrast, even if the incident angle of the light is different, the colored glass filter can exert the same filtering performance, thereby achieving the above-mentioned effect.

[0283] Fig.28 Shows the generation and Fig. 27 Other structures with the same effect. The focusing lens array device 83 as a component of the sub-device 85 is made of transparent materials such as glass and resin as described above, while the V-groove array device 84 is made of opaque materials. By forming pinholes 87 as through holes at the positions where the V-groove array device 84 intersects with the optical axes of each focusing lens 2, the V-groove array device 84 acts as a pinhole array. By adopting such a structure, it is possible to Fig. 27 The sub-device 85 can be made more simply in this case.

[0284] In addition, the present invention is not limited to the above-mentioned embodiments, and includes various modified examples. For example, the above-mentioned embodiments are described in detail in order to explain the present invention in an easy-to-understand manner, and are not limited to having all the structures described. In addition, a part of the structure of a certain embodiment can be replaced with the structure of another embodiment, and the structure of another embodiment can also be added to the structure of a certain embodiment. In addition, other structures can be added, deleted, or replaced with respect to a part of the structure of each embodiment.

[0285] Explanation of symbols

[0286] 1—light point, 2—condensing lens, 3, 3'—parallel light beam, 4, 5—light point, 7—light point image, 8—condensing lens array, 11—two-dimensional color sensor, 17-20—color separation mirror, 30—two-dimensional sensor, 31—transmission diffraction grating, 33—recondensing lens, 37—two-dimensional sensor, 41—wavelength dispersion image, 43—prism, 45, 48—light point image, 49—capillary, 53—laser source, 71—light point, 72—pinhole, 74—pinhole image, 75—light point, 76—light point image, 77-79—optical path length adjustment element, 80—light distribution, 83—condensing lens array device, 84—V-groove array device, 87—pinhole.

Claims

1. A luminescence detection device, characterized in that: have: A capillary array formed by arranging a plurality of capillaries on a plane; A device supporting the capillary array; A condenser lens array, which is arranged with M condenser lenses and is integrated with the above device, wherein the M condenser lenses individually condense the light emitted from a light-emitting point array having M light-emitting points arranged therein to form a light beam, wherein the light-emitting point array is formed by the light-emitting points from the capillary array, and M≥2; and at least one sensor that causes M of the above light beams to be incident in parallel, The average value of the effective diameters of the M light-emitting points is set as d. The average focal length of the M focusing lenses is set to f. The average value of the intervals between the M focusing lenses is set to p. The average value of the maximum optical path length between the M condenser lenses and the at least one sensor is set to g. At this time, At least d, f, p, and g associated with any one of the above M light-emitting points satisfy the following relationship: f≥1 / ((2×p) / (1.27×d)+1)×g.

2. The luminescence detection device according to claim 1, characterized in that: The device further comprises a pinhole array, which is disposed between the capillary array and the condensing lens array and is formed by arranging M pinholes so as to coincide with the positions of the M condensing lenses.

3. The luminescence detection device according to claim 1 or 2, characterized in that: Satisfies f≥1 / (p / d+1)×g.

4. The luminescence detection device according to claim 1 or 2, characterized in that: Satisfies f≤2×p.

5. The luminescence detection device according to claim 1 or 2, characterized in that: Satisfies f≤p.

6. The luminescence detection device according to claim 1 or 2, characterized in that: A color separation mirror array is provided, wherein the color separation mirror array is arranged in the optical middle of the above-mentioned focusing lens array and the above-mentioned sensor, and N color separation mirrors numbered 1, 2, ..., N are arranged in a numbered order in a first direction, wherein N ≥ 2, The normal vectors of the front surfaces of the N dichroic reflectors are composed of the sum of the positive component in the first direction and the negative component in the second direction perpendicular to the first direction. The above N normal vectors are roughly parallel to each other. The optical axes of the M focusing lenses are substantially parallel to the second direction. The arrangement direction of the M condensing lenses is substantially parallel to a third direction perpendicular to both the first direction and the second direction.

7. The luminescence detection device according to claim 6, characterized in that: The M light beams are incident in parallel along the second direction into the dichroic reflector array. The dichroic reflector array emits M×N split light beams along the second direction by splitting the M light beams into N different light beams in the first direction, The M×N divided light beams are incident on the sensor in parallel and are detected collectively.

8. The luminescence detection device according to claim 6, characterized in that: The average effective diameter of the M focusing lenses is set as D. The average value of the angles formed by the N normal vectors relative to the direction opposite to the second direction is set to θ0, where 0≤θ0≤90°. The average refractive index of the base material of the N dichroic reflectors is set to n0. The average value of the width of the base material of the N dichroic reflectors is set to α. The average thickness of the base material of the N dichroic reflectors is set to β. Let the average value of the intervals between the N dichroic reflectors be x, The average value of the distance by which the end of the dichroic reflector numbered n in the second direction deviates in the opposite direction to the end of the dichroic reflector numbered n-1 in the second direction is yz, where 2≤n≤N. In this case, d, f, D, p, g, θ0, N, n0, α, β, x, and yz satisfy a predetermined relationship so that the M light emissions can be detected using the color separation mirror.

9. The luminescence detection device according to claim 8, characterized in that: Let θ1 = sin -1 (1 / n0×sinθ0), θ2=sin -1 (1 / n0×sin(90°-θ0)), a W =cosθ0, b W =-cosθ0×tanθ1, a L =(N-1)×cosθ0+sinθ0, b L =(N-2) / cosθ0×(2×sin(90°-θ0-θ2)+1-sin(θ0+θ2))+(N-2)×sinθ0+2×cosθ0, d'=(g-f) / f×d, Satisfies (sinθ0-b W / a W × cosθ0) × β + cosθ0 / a W ×D≤x, and (sinθ0-b W / a W ×cosθ0)×β+cosθ0 / a W ×d’≤x≤(sinθ0-b L / a L ×cosθ0)×β+cosθ0 / a L ×g。 10. The luminescence detection device according to claim 8, characterized in that: Let θ2 = sin -1 (1 / n0×sin(90°-θ0)), Satisfies 0≤yz≤2×sin(90°-θ0-θ2) / cosθ2×β.

11. The luminescence detection device according to claim 8, characterized in that: Let θ2 = sin -1 (1 / n0×sin(90°-θ0)), When n = 2, 0 ≤ yz ≤ 2 × cos θ0 × β is satisfied. And when 3≤n≤N, 0≤yz≤2×sin(90°-θ0-θ2) / cosθ2×β is satisfied.

12. The luminescence detection device according to claim 8, characterized in that: It satisfies cosθ0×α≤x≤cosθ0×α+2×sinθ0×β.

13. The luminescence detection device according to claim 7, characterized in that: An optical path length adjusting element is provided on at least a part of the optical paths of the M×N divided light beams.

14. The luminescence detection device according to claim 1 or 2, characterized in that: The M condenser lenses are two-dimensionally arranged in a plane parallel to the sensor surface of the sensor.

15. The luminescence detection device according to claim 1 or 2, characterized in that: The sensor is a color sensor in which a plurality of types of pixels for recognizing different wavelength bands are arranged.

16. The luminescence detection device according to claim 1 or 2, characterized in that: A mechanism is provided for moving at least the relative position of the condenser lens array with respect to the M light-emitting points.

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