JP2019074536A - Multi-color detection device - Google Patents

Abstract

To provide a small size device for detecting multiple colors of light emitted from an array of light emission points capable of detecting multiple colors with high sensitivity and low crosstalk at lower manufacturing cost.SOLUTION: The device is for detecting multiple colors of light emitted from an array of light emission points using an individual condenser lens and a common wavelength dispersion element. The multi-color detection device comprises: a condenser lens array 17 in which multiple condenser lenses 18 are arranged for independently forming parallel luminous flux respectively from light 16 emitted from respective emission points of the array of light emission points where multiple light emission points 15 are arranged; at least one common spectroscopic element (common transmission grating 21) on which the parallel luminous fluxes 19 are incident in parallel; and at least one common sensor surface 27 on which light fluxes 22 separated by the spectroscopic element are incident in parallel.SELECTED DRAWING: Figure 7

Description

  The present invention irradiates light such as a laser beam to a plurality of channels provided inside a plurality of capillaries or microchips, and highly sensitive to fluorescence, phosphorescence, scattered light, etc. emitted from substances present inside the capillaries or channels. Relates to a multicolor detection device for detecting

  Capillary performing batchwise decipherment of base sequences of different DNA samples by individual capillaries by parallel processing of electrophoresis analysis by a plurality of capillaries (glass capillaries with an outer diameter of 100 to 400 μm and an inner diameter of 25 to 100 μm) filled with a separation medium Array DNA sequencers are widely used. This mechanism is described next. A commercially available capillary forms a polyimide coating film on the outer surface to give flexibility. A portion where the electrophoresis path length of each capillary is constant, for example, a position near a distance of 30 cm from the sample injection end of the capillary is aligned on the same plane with the coating film removed, and the laser beam is arranged in the above capillary arrangement By irradiating from the side direction along the plane, multiple capillaries are irradiated simultaneously. Hereinafter, in the present specification, the above-mentioned capillary array plane may be simply referred to as an array plane. The fluorescence labeled DNA which electrophoreses the inside of each of the above-mentioned capillaries receives fluorescence excitation upon receiving a laser beam and emits fluorescence. Here, the DNA is dyed into four-color phosphors according to the A, C, G, T base species. As a result, the laser irradiation position of each capillary becomes a light emitting point, and a plurality of light emitting points are arranged on a straight line at an interval p. Hereinafter, this is called a light emitting point array. Assuming that the number of light emitting points (the number of capillaries) is n, the total width W of the light emitting point array is W = p * (n−1). For example, when p = 0.36 mm and n = 24, W = 8.28 mm. The fluorescence detection device collectively detects each light emission from the light emission point array while dispersing it. This device configuration is shown in FIG. 3 of Patent Document 1.

  First, each light emission is collimated by a common condenser lens. Hereinafter, the expression “common” is used in the sense that one optical element is used for a plurality of (n) light emitting points (corresponding to n: 1). On the contrary, the expression "individual" is used in the sense that one optical element is used for one light emitting point (1: 1 correspondence). Here, assuming that the focal length of the common condenser lens is f and the effective diameter is D1, W <f, W <D1. For example, f = 50 mm and D1 = 36 mm. Next, a parallel beam is passed through a long pass filter to cut the wavelength of the laser beam, and the common transmission type diffraction grating is further transmitted to make the major axis direction of each capillary, ie, the arrangement direction of the light emission point array and the light of the common condenser lens Wavelength dispersion is performed in the direction orthogonal to both axes. Here, assuming that the effective diameter of the common transmission type diffraction grating is DG, in order not to reduce the detection efficiency, it is necessary to satisfy D1 ≦ DG. For example, DG = 50 mm. Subsequently, each collimated light beam is imaged on a two-dimensional sensor by a common imaging lens. Here, assuming that the effective diameter of the common imaging lens is D2, in order not to lower the detection efficiency, it is necessary to satisfy D1 ≦ D2. For example, D2 = 36 mm. As described above, the wavelength dispersion spectrum of each light emission from the light emitting point array can be acquired at once. Finally, the time change of the fluorescence intensity of four colors is determined by analyzing the time change of each wavelength dispersion spectrum, and the order of the base species, that is, the base sequence is determined.

  Another means of simultaneously detecting four colors of fluorescence is shown in FIG. 2 of Non-Patent Document 1. First, light emitted from one light emitting region is collimated by one condensing lens (here, an objective lens). Here, assuming that the total width of the light emitting region is W, the focal length of the objective lens is f, and the effective diameter is D1, W <f, W <D1. The objective used is an Olympus UPLSAPO 60 × W, W = 0.44 mm, f = 3 mm, D1 = 20 mm. Next, the parallel light flux is divided into four parallel light fluxes of four colors by one set of three types of dichroic mirrors. Subsequently, each collimated light beam is imaged on each of four two-dimensional sensors by one set of four imaging lenses. Here, assuming that the effective diameter of each imaging lens is D2, in order not to lower the detection efficiency, it is necessary to satisfy D1 ≦ D2. According to the above, it is possible to acquire four-color divided images of the light emitting area collectively.

  On the other hand, another means for simultaneously detecting the light emission from the light emitting point array is shown in FIG. 1 of Patent Document 2. First, each light emission from the light emitting point array is collimated by an individual condensing lens array. Here, assuming that the distance between light emitting points is p and the number of light emitting points is n, the total width of the light emitting point array is W = p * (n-1), and the effective diameter of each condenser lens is D1. It is <W. Further, by setting D1 <p, it is possible to obtain an individual condensing lens array in which each condensing lens is arranged in a straight line. Next, each collimated light beam is incident on each individual sensor of an individual sensor array. Thus, the light emission intensity from the light emission point array can be acquired collectively.

JP 2007-171214 A JP 2011-59095 A

Rev Sci Instrum., 2011 Feb; 82 (2): 023701.

In the fluorescence detection device of Patent Document 1, the light collection efficiency of light emission from each light emitting point (light collection efficiency by common light collecting lens), detection efficiency (light collection efficiency, transmittance of long pass filter, diffraction efficiency of diffraction grating, etc. Based on the above, the total utilization efficiency of the light emission contributing to the fluorescence detection by the sensor is high, and the spectral accuracy by the diffraction grating is also high. However, since there is a relationship of W <f, W <D1 ≦ D2 including two common lenses (using a camera lens), if W is constant, the overall size of the device is very large, and the manufacturing cost of the device is High is the problem. For example, in the case of f = 50 mm, D1 = 36 mm, D2 = 36 mm, the overall size of the fluorescence detection device is larger than the volume (1.6 × 10 ⁶ mm ³ ) of a cylinder with a diameter of 100 mm and a height of 200 mm. In this specification, the overall size of the fluorescence detection apparatus is expressed by the occupied volume of the optical system from the light emission point to the imaging point, and the occupied volume of the sensor itself is not included. Since W << f and W <D1 can not be satisfied (a huge camera lens is required to realize this), detection of the light emitting point near the optical axis (light emitting point located near the center of the light emitting point array) Compared with the efficiency, the detection efficiency of the light emitting point (light emitting point located near the end of the light emitting point array) away from the optical axis is lowered, and there is a problem that the sensitivity varies for each light emitting point.

  However, to solve these problems, ie, to miniaturize and reduce the cost of a device that simultaneously detects four colors of light emitted from a light emitting point array and to reduce the cost variation of each light emission, I did not come. If the size of the fluorescence detection apparatus can be reduced, the capillary array DNA sequencer can be installed in a small area, can be carried around, or the usability can be improved. In addition, the number of parts of the fluorescence detection device is reduced, and the size of each part is reduced, whereby the manufacturing cost is reduced. Furthermore, by reducing the sensitivity variation of each light emitting point, it becomes possible to make a quantitative comparison of the samples analyzed by each capillary, and it is possible to improve the total sensitivity and dynamic range of the light emitting point array. As a result of these, the capillary array DNA sequencer can further spread and contribute to the world.

  The fluorescence detection apparatus shown in Non-Patent Document 1 can be used to perform simultaneous fluorescence detection of four-color light emission from a similar light emitting point array. However, with the objective lens used here, since W = 0.44 mm, for example, only a part of the full width W = 8.28 mm of the light emitting point array can be detected. Therefore, instead of the objective lens and the four individual imaging lenses, a common condenser lens and four common imaging lenses are used as in the capillary array DNA sequencer. At this time, assuming that the effective diameters of the three types of dichroic mirrors are DM, they are arranged to be inclined 45 ° with respect to the parallel light flux, and in order not to reduce the detection efficiency, it is necessary to satisfy √2 × D1 ≦ DM. For example, DM = 71 mm. Therefore, even if the four cameras are not included, the overall size of the fluorescence detection device becomes larger than in the case of Patent Document 1, and the manufacturing cost increases accordingly. In addition to this, the space occupied by the four cameras is large and the cost is very high. The problem of sensitivity variation for each light emitting point also remains.

  On the other hand, using the fluorescence detection device disclosed in Patent Document 2 may reduce the device size because D1 <W, but the problem is that it corresponds to only one-color fluorescence detection. Therefore, following Patent Document 1 and combining with wavelength dispersion by a diffraction grating will be considered. Light emitted from n light emitting points is collimated by n individual condenser lenses, each of which is transmitted through n individual transmission type diffraction gratings for wavelength dispersion, and n n imaging lenses Image on a one-dimensional or two-dimensional individual sensor. That is, the fluorescence detection device of Patent Document 1 is miniaturized, and n pieces of the fluorescence detection device are arranged in parallel. Here, since D1 <p, p = 0.36 mm, for example, D1 = 0.3 mm. The effective diameter DG of the transmission type diffraction grating does not receive steric hindrance with the adjacent diffraction grating simultaneously with D1 ≦ DG, so it is necessary to set DG <p, for example, it may be DG = 0.3 mm. The above fluorescence detection device can be miniaturized as compared with the case of Patent Document 1, but it is difficult to produce n pieces of fine optical components and to arrange each in a predetermined position. Yes, and the manufacturing cost is higher. In addition, it is difficult to fabricate a transmission grating of DG = 0.3 mm.

  Next, according to Non-Patent Document 2, consider combining with three types of dichroic mirrors. Light emitted from n light emitting points is converted into parallel light beams by n individual condenser lenses, and n parallel light beams are divided into 4 groups of 4 colors using n groups × 3 types of individual dichroic mirrors. The divided parallel light is formed, and images are formed on n sets × 4 individual sensors by n sets × 4 individual imaging lenses. That is, the fluorescence detection device of Non-Patent Document 1 is miniaturized, and n pieces of the fluorescence detection device are arranged in parallel. Here, since D1 <p, p = 0.36 mm, for example, D1 = 0.25 mm. The effective diameter DM of each dichroic mirror does not receive a steric hindrance with the adjacent dichroic mirror at the same time as 2 2 × D 1, DM, so it is necessary to set DM <p. For example, if DM = 0.35 mm good. Although the above-mentioned fluorescence detection device reduces the variation in sensitivity due to the light emission point compared to the case of Patent Document 1, n pieces or n sets of fine optical components are prepared, and each of them is set to a predetermined value. It is difficult to arrange in position, and the manufacturing cost is higher. In addition, it is difficult to arrange n four-divided images while avoiding steric hindrance. Furthermore, it is also difficult to fabricate a dichroic mirror with DM = 0.35 mm.

  In the above, four-color fluorescence detection has been described on the assumption that the present invention is applied to a fluorescence detector of a capillary array DNA sequencer, but the problem is not limited to capillary or four-color fluorescence detection. Is common to the case of detecting multicolor emission of two or more colors.

  The multicolor detection device according to the present invention comprises a condenser lens array in which a plurality of condenser lenses are arrayed for individually making parallel luminous flux emitted from each luminous point of a luminous point array in which a plurality of luminous points are arrayed It has a common and at least one dispersive element in which the luminous fluxes are respectively incident in parallel and a common and at least one sensor in which the luminous fluxes separated by the dispersive elements are respectively incident in parallel.

  A diffraction grating, a prism, or a dichroic mirror can be used as the spectral element.

  In the multicolor detection device according to the present invention, there is provided a condenser lens array in which a plurality of condenser lenses are arranged to individually make parallel luminous flux emitted from each luminous point of the luminous point array in which a plurality of luminous points are arrayed; Common and at least one color sensor, in which the parallel luminous fluxes are respectively incident in parallel.

In the multicolor detection device according to the present invention, there is provided a condenser lens array in which a plurality of condenser lenses are arranged to individually make parallel luminous flux emitted from each luminous point of the luminous point array in which a plurality of luminous points are arrayed; The parallel luminous flux has a common and at least one sensor, each of which is incident in parallel, the average of the effective diameter of the light emitting point is d, the average of the focal length of the focusing lens is f, the effective diameter of the focusing lens Let D be the average of and g be the average of the optical distance between the condenser lens and the sensor,
f ≦ −0.20 * (d / D) * g + 2.8 * D
Satisfy.

Further, the multicolor detection device according to the present invention comprises a condenser lens array in which condenser lenses are arrayed individually to make light emitted from each luminous point of the luminous point array in which plural luminous points are arrayed into parallel luminous flux, and the parallel With common and at least one sensor, in which the luminous fluxes are respectively incident in parallel, the average of the effective diameter of the light emission point is d, the average of the arrangement interval of the light emission points is p, the condenser lens and the optical of the sensor Let g be the average of various distances,
f ≧ 0.95 * (d / p) * g
Satisfy.

Further, the multicolor detection device according to the present invention comprises a condenser lens array in which condenser lenses are arrayed individually to make light emitted from each luminous point of the luminous point array in which plural luminous points are arrayed into parallel luminous flux, and the parallel It has an imaging lens array in which a plurality of imaging lenses individually arraying luminous fluxes, and a common and at least one sensor into which the luminous fluxes are incident in parallel, and the luminous point is effective The average of the diameter d, the average of the focal length of the focusing lens f, the average of the effective diameter of the focusing lens D, the optical distance between the focusing lens and the imaging lens corresponding to the focusing lens Let g be the average of
f ≦ −0.20 * (d / D) * g + 2.8 * D
Satisfy.

Further, the multicolor detection device according to the present invention comprises a condenser lens array in which condenser lenses are arrayed individually to make light emitted from each luminous point of the luminous point array in which plural luminous points are arrayed into parallel luminous flux, and the parallel It has an imaging lens array in which a plurality of imaging lenses individually arraying luminous fluxes, and a common and at least one sensor into which the luminous fluxes are incident in parallel, and the luminous point is effective D of average diameter, p of average arrangement distance of light emitting points, f of average focal distance of condenser lens, optical distance between the condenser lens and the imaging lens corresponding to the condenser lens Let g be the average,
f ≧ 0.95 * (d / p) * g
Satisfy.

  Further, in the device according to the present invention, a channel array in which at least a part of a plurality of channels are arranged on the same plane, and a condensing lens in which condensing lenses individually making parallel light beams emitted from the respective channels of the channel array are arranged. The lens array is integrated.

  The plurality of channels may be inside the plurality of capillaries or may be formed inside the microchip.

  According to the present invention, it is possible to miniaturize a device for performing multicolor detection of light emission from a light emitting point array, and to miniaturize the overall size of various devices using this. Therefore, the space for placing the device can be reduced, the device can be carried, and the usability of the device is improved. Further, the number of parts constituting the apparatus can be reduced, and the manufacturing cost can be reduced by miniaturizing the parts themselves.

  Problems, configurations, and effects other than those described above will be clarified by the description of the embodiments below.

The schematic diagram which shows the example of a structure of the fluorescence detection apparatus which parallelizes each light emission from a light emission point array with an individual condensing lens, makes it inject into a sensor area, and is detected. The figure which showed the relationship of g and relative detection light quantity by making f as a parameter. The figure which showed the relationship of g and f from which relative detection light quantity becomes 50% or more by setting D as a parameter. The figure which showed the relationship of g and f from which crosstalk signal strength ratio becomes 25% or less by making p into a parameter. The figure which showed the relationship of g and f from which a relative detection light quantity becomes 50% or more, and crosstalk signal strength ratio becomes 25% or less by using D and p as a parameter. The schematic diagram which shows the apparatus structural example of a capillary array DNA sequencer. FIG. 5 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting the light emission from the light emitting point array with an individual condensing lens and a common wavelength dispersion element in multiple colors. FIG. 7 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting light emitted from the light emitting point array by a multicolor detection by an individual condensing lens, a common wavelength dispersion element, and a sensor arranged perpendicularly to the optical axis of the condensing lens. FIG. 7 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting light emitted from the light emitting point array by a multicolor detection by an individual condensing lens, a common concave reflection type diffraction grating, and a sensor arranged perpendicular to the optical axis of the condensing lens. FIG. 5 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting the light emission from the light emission point array with an individual condensing lens, a common dichroic mirror set, and a multicolor sensor. FIG. 7 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting light emission from the light emission point array with an individual condensing lens, a common dichroic mirror set, and a sensor as polychromatic in the same manner as wavelength dispersion. The cross-sectional schematic diagram which shows the apparatus structural example which detects multiple colors of light emission from a light emission point array with an individual condensing lens and a color sensor. The cross-sectional schematic diagram which shows the structural example of the device which integrated the V groove | channel array which arranges several capillaries, and the separate condensing lens array. The cross-sectional schematic diagram which shows the structural example of the device which adhere | attached the separate condensing lens to several capillaries, respectively. The cross-sectional schematic diagram which shows the structural example of the device which the microchip which has a multi-channel, and the separate condensing lens array were integrated. A schematic cross-sectional view showing an example of a device configuration in which light emission from a light emitting point array by a device in which a microchip having a multi-channel and an individual condensing lens array are integrated and individual LED illumination is detected by a common dichroic mirror set and a sensor .

  The present invention provides a means for miniaturizing and reducing the cost of a device for simultaneously detecting four color light emissions from a light emitting point array while detecting the variations in sensitivity of each light emission. First, an overview of the present invention is given.

  First, each light emission from the light emitting point array is collimated by an individual condensing lens array. In this specification, although the expression “parallel light flux” is often used, the constituent light elements do not necessarily mean light fluxes parallel to one another in a strict sense, but the light elements of light emission emitted in all directions from light emission points This means that the angle is at least reduced by the collecting lens and is approaching zero. Here, assuming that the average of the spacing of the light emitting points is p, the number of light emitting points and the number of individual condenser lenses are n, the total width of the light emitting point array is W = p * (n-1). Assuming that the average of the focal lengths of the focusing lenses is f and the average of the effective diameters is D1, D1 <W. Further, by setting D1 <p, it is possible to make a condensing lens array in which the respective condensing lenses are arranged in a straight line. For example, f = 1 mm and D1 = 0.3 mm for p = 0.36 mm, n = 24 and W = 8.28 mm.

  Next, each parallel beam is transmitted through a spectral element, for example, a single common transmission diffraction grating to disperse the wavelength. Here, assuming that the effective diameter in the arrangement direction of the light emitting point array of the transmission type diffraction grating is DG1 and the effective diameter in the major axis direction of each capillary of the transmission type diffraction grating is DG2, the detection efficiency is not reduced. It is assumed that DG1 and D1 ≦ DG2. For example, DG1 = 10 mm and DG2 = 1 mm. At this time, the 24 parallel light beams separated from each other are incident on different portions of one common transmission type diffraction grating, and each receives wavelength dispersion in parallel. The diameter of each parallel beam is D1 = 0.3 mm, which is sufficiently large as compared with the grating constant of the diffraction grating, so that each parallel beam can receive good wavelength dispersion. The direction of wavelength dispersion is the direction of the major axis of each capillary, that is, the direction perpendicular to both the arrangement direction of the light emitting point array and the optical axis of each condenser lens.

  Subsequently, each collimated light beam subjected to wavelength dispersion is imaged on one common two-dimensional sensor by n individual imaging lenses. The average D2 of the effective diameter of each imaging lens needs to be D1 ≦ D2 in order not to reduce the detection efficiency. For example, D2 = 0.3 mm. As described above, the four-color fluorescence detection of each light emission from the light emitting point array can be performed collectively.

  According to the above fluorescence detection device, not only the device size is miniaturized as compared with the case of Patent Document 1, but also the device configuration is simplified by making the diffraction grating and the sensor common to a plurality of light emitting points. , Implementation is facilitated. The overall size of the fluorescence detection device can be smaller than a cylinder with a diameter of 10 mm and a height of 20 mm. Further, although the diffraction grating and the sensor are shared, the fluorescence detection optical system and the detection efficiency are equivalent for each light emitting point, and it is possible to reduce the sensitivity variation. The above can similarly solve the problem even if the transmission type diffraction grating is replaced with a wavelength dispersive prism.

  Another aspect of the present invention will be described. First, each light emission from the light emitting point array is collimated by an individual condensing lens array. Here, assuming that the average of the spacings of the light emitting points is p, the number of light emitting points and the number of condenser lenses are n, the total width of the light emitting point array is W = p * (n-1). Assuming that the average of the focal lengths of the focusing lenses is f and the average of the effective diameters is D1, D1 <W. Further, by setting D1 <p, it is possible to make a condensing lens array in which the respective condensing lenses are arranged in a straight line. For example, f = 1 mm and D1 = 0.3 mm for p = 0.36 mm, n = 24 and W = 8.28 mm.

  Next, for each parallel beam, one set of three common dichroic mirrors and one total reflection mirror both in the major axis direction of each capillary, that is, both in the array direction of the light emission point array and in the optical axis of each condenser lens. By arranging in parallel in the orthogonal direction, n sets of four-color divided parallel luminous fluxes are obtained, and these parallel luminous fluxes are parallel to the direction perpendicular to the arrangement plane of the capillary array, that is, the optical axis of each condenser lens Advance in the direction. Assuming that the average of the effective diameters in the arrangement direction of the light emitting point arrays of each dichroic mirror and total reflection mirror is DM1 and the average of the effective diameters in the orthogonal direction is DM2, the detection efficiency is not reduced. It is assumed that 2 × D1 ≦ DM2. For example, DM1 = 10 mm and DM2 = 1 mm. At this time, the 24 parallel light fluxes separated from each other are made incident on different portions of one dichroic mirror for each type, and each of them is split into a transmitted light and a reflected light. Since one dichroic mirror can obtain uniform performance at any place, each collimated light beam can receive good spectral separation. The total reflection mirror may be replaced by a dichroic mirror.

  Subsequently, n sets of four color-divided parallel luminous fluxes are incident on one common two-dimensional sensor without imaging. In the case of spectral separation by wavelength dispersion by a diffraction grating or a prism, as described above, desired spectral accuracy can not be obtained unless an image of a wavelength-dispersed parallel light beam is imaged using an imaging lens. On the other hand, the imaging lens can be omitted because it is not always necessary in the case of splitting by a dichroic mirror. As described above, the four-color fluorescence detection of each light emission from the light emitting point array can be performed collectively.

  According to the above fluorescence detection device, not only the device size is miniaturized as compared with the case of Patent Document 1, but the device configuration is simplified by sharing the dichroic mirror and the sensor for a plurality of light emitting points. , Implementation is facilitated. The overall size of the fluorescence detection device can be smaller than a cylinder with a diameter of 10 mm and a height of 10 mm. Moreover, although the diffraction grating and the sensor are shared, the fluorescence detection system and the detection efficiency are equivalent for each light emitting point, and it is possible to reduce the sensitivity variation.

  Another aspect of the present invention will be described. First, each light emission from the light emitting point array is collimated by an individual condensing lens array. Here, assuming that the average of the spacings of the light emitting points is p, the number of light emitting points and the number of condenser lenses are n, the total width of the light emitting point array is W = p * (n-1). Assuming that the average of the focal lengths of the focusing lenses is f and the average of the effective diameters is D1, D1 <W. Further, by setting D1 <p, it is possible to make a condensing lens array in which the respective condensing lenses are arranged in a straight line. For example, f = 1 mm and D1 = 0.35 mm for p = 0.36 mm, n = 24 and W = 8.28 mm.

  Subsequently, each parallel luminous flux is made to enter a single two-plate common two-dimensional color sensor without imaging. In the color sensor, at least four kinds of pixels respectively identifying four colors are arranged in a large number on the two-dimensional sensor surface, or one type of each pixel in which a large number is arranged is perpendicular to the two-dimensional sensor surface ( Four colors are identified in the traveling direction of the incident light. Here, assuming that the average of the diameter of each pixel is S, it is necessary to satisfy S <D1. The average diameter of each collimated beam is D1 = 0.35 mm. On the other hand, using a sensor in which four types of pixels for identifying four colors are arrayed and S = 0.05 mm, each parallel luminous flux is detected at about 40 pixels on the color sensor. At this time, since detection is performed with about 10 pixels per one type of pixel for identifying one color, by integrating these, variation for each color can be reduced, and highly accurate spectroscopy can be performed.

  On the other hand, if each collimated beam is imaged by an individual condensing lens array, and the diameter of the imaging spot becomes 0.05 mm, for example, it will be detected only at about 1 pixel on the color sensor. , Good spectroscopy becomes impossible. That is, in this aspect, not intentionally using the imaging lens not only contributes to downsizing of the apparatus but also contributes to improving the spectral accuracy. As described above, the four-color fluorescence detection of each light emission from the light emitting point array can be performed collectively.

  According to the above fluorescence detection device, not only the device size is miniaturized as compared with the case of Patent Document 1, but also the device configuration is extremely simple. The overall size of the fluorescence detection device can be smaller than a cylinder with a diameter of 10 mm and a height of 5 mm. Further, the fluorescence detection system and the detection efficiency are equivalent for each light emitting point, and it is possible to reduce the sensitivity variation.

  In the above, four-color fluorescence detection has been described on the assumption that the present invention is applied to a fluorescence detector of a capillary array DNA sequencer, but the solution to the problem is not limited to capillary or four-color fluorescence detection. It is common to multicolor luminescence detection of two or more colors about luminescence from a point array.

  Hereinafter, the present invention will be described in detail with reference to the drawings.

  Although the size of each light emitting point of the light emitting point array targeted by the present invention is small, it has a finite size and can not be ignored when the fluorescence detection device is miniaturized. FIG. 1 is a schematic view showing a configuration example of a fluorescence detection apparatus which collimates each light emission from a light emission point array by individual condenser lenses and makes the light incident on a sensor area for detection. FIG. 1 shows that the light emission from the light emitting point 15 of the average effective diameter d is collimated by the individual focusing lens 18 of the average focal length f and the average effective diameter D, and the average distance g from the individual focusing lens 18 The configuration is shown in which the sensor region 28 of the average effective diameter D is made incident and detected. The average effective diameter D of the sensor area 28 does not necessarily indicate the overall size of the sensor, and part of a larger sensor can be considered as an area allocated for detection of the light emitting point 15. Also, although not shown in FIG. 1, in the case of a fluorescence detection apparatus in which each collimated light beam is refocused or imaged by an individual imaging lens and then incident on a sensor area to be detected, in the following discussion, The sensor area may be replaced with an individual imaging lens, and the average distance between the individual condensing lens and the individual imaging lens may be g.

First, focus on the light emitting point 15 on the left side of FIG. Light 29 emitted from the center of the light emitting point 15 is collimated by the condenser lens 18 to form a parallel light beam 30, and a spot 31 is formed on the sensor area 28, and the sensor area 28 and the spot 31 coincide. Assuming that D is constant, the amount of light detected at this time increases with the light receiving angle θ1 as f is smaller. More precisely, when F = f / D, the detected light quantity increases in proportion to 1 / F ² . On the other hand, the spot 34 of the parallel luminous flux 33 of the light 32 emitted from the left end of the light emitting point 15 is shifted to the right from the sensor area 28. That is, although all the spots 31 are detected, the spots 34 are detected only at a ratio overlapping the spots 31. The larger the overlap, the larger the amount of light detected for the entire area of the light emitting point. For this purpose, the angle θ2 between the optical axis of the parallel light beam 30 and the optical axis of the parallel light beam 33 may be small, and for this purpose, assuming that d is constant, the larger the f, the better. As described above, in order to increase the amount of light detected at the light emitting point 15, there is a trade-off between the surface where it is better to reduce f and the surface where it is better to increase f. A study of the best has not been done so far. Therefore, the conditions of f and g for increasing the amount of light detected at the light emitting point 15 will be clarified next.

In order to evaluate the amount of detected light, the fluorescence detection device shown in FIG. 3 of Patent Document 1 is used as a reference. In a typical example of this fluorescence detection apparatus, the focal length of the common condenser lens is f = 50 mm, and the effective diameter is D1 ≧ 25 mm. The brightness of this lens is F = f / D1 ≦ 2.0. Therefore, when a condenser lens of F ₀ = 2.0 is used, the light emission from the infinitely small-sized light emission point located at the focal point is collimated by this lens, and all the light quantities are detected by the sensor without loss The detected light quantity is taken as a reference (100%). Hereinafter, the amount of light detected for a light emitting point of an infinitesimal small size is evaluated based on the amount of light detected relative to the above reference. In addition, it is considered that the light emitting point of finite size of the average effective diameter d is composed of many light emitting points of infinitely small size. In the present specification, the "light-emitting point of finite size" is simply referred to as "light-emitting point", and the "light-emitting point of infinitesimal size" is referred to as the "light-emitting point of infinitesimal size" each time. The relative detection light amount of the light emission point is an average of the relative detection light amounts of a large number of infinitely small-sized light emission points constituting the light emission point. For example, in the above example, when the condenser lens is replaced with F = 1.4, the light collection efficiency is (F ₀ / F) ² = 2.0 times, so the relative detection of the light emission point of infinitely small size The light amount is 200%. However, it is assumed that the total light quantity emitted in all directions from the light emitting point is constant, and the light emission density inside the light emitting point is spatially uniform. Moreover, in the typical example of the present fluorescence detection device, the spacing of the light emitting points of the light emitting point array is p = 0.36 mm, the number of light emitting points is n = 24, and the total width of the light emitting point array is W = p * (n-1) = 8.28 mm, and the light emitting point located at the center of the light emitting point array is located near the focal point of the lens, so the relative detection light amount is almost 100%, but the light emitting point located at the end of the light emitting point array is the lens The relative detection light amount decreases to be about 50% because it is away from the focal point. Therefore, in the present invention, it is an object of the present invention to set the multicolor detection sensitivity of each light emitting point to be equal to or more than the conventional one so that the relative detection light quantity of each light emitting point is 50% or more.

  FIG. 2 is a diagram showing the result of calculating the relationship between g and the relative amount of detected light with f as a parameter in the configuration shown in FIG. Here, the average effective diameter of the light emitting point 15 is d = 0.05 mm. Further, the average effective diameter of the individual condensing lens 18 is D = 0.5 mm. The relative detection light quantity was calculated in consideration of the lens brightness F = f / 0.05. The light-emitting point 15 of effective diameter d = 0.05 mm is composed of approximately 500 infinite-small-size light-emitting points spaced by 0.1 μm, and for each of the infinite-small-size light-emitting points, the spots 31 and 34 of FIG. The relative detection light amount was calculated according to the same concept as the overlapping area ratio, and the relative detection light amount of the light emitting point 15 was determined by averaging them. As a result, it was found for the first time that the relative detection light quantity becomes larger as f is smaller and g is smaller. The effect of increasing the relative detection light quantity of the infinitesimally-sized light emitting point located at the center of the light emitting point 15 by making f small (F small and θ1 large) increases f large (small θ2) By doing this, it is shown that the effect of increasing the above-mentioned overlapping area ratio is greater. It also shows that the effect of increasing the overlapping area ratio is large by decreasing g for any f.

FIG. 3 shows the relationship between g and f that satisfies the condition of 50% or more of the relative detection light amount based on the calculation result of FIG. 2 as a graph of the horizontal axis g and the vertical axis f. Here, D is a parameter. It was found that the relative detection light amount would be 50% or more if g and f were in the region below the straight line having a negative slope, even if D was any value. The larger the value of D, the larger the intercept of the vertical line of the boundary line, and the smaller the inclination, so the area satisfying the condition becomes larger. As a result of detailed analysis, the region that satisfies the condition is generally
f ≦ −0.20 * (d / D) * g + 2.8 * D (1)
It turned out that it can express with. Similar to the result of FIG. 2, the relative detection light amount increases as f and g decrease, that is, as it approaches the origin of FIG. However, since there are various physical restrictions in practice, it is preferable to set appropriate f and g from the area shown in FIG.

  On the other hand, in FIG. 1, similarly to the light emission point 15 on the left side, the light emission 35 from the center of the light emission point 15 on the right side is made a parallel light beam 36 by the condenser lens 18 and its spot 37 coincides with the sensor area 28 It is detected without any loss. However, the spot 34 of the parallel luminous flux 33 of the light emission 32 from the left end of the light emission point 15 on the left side may be shifted to the right from the sensor area and overlap the sensor area of the light emission point on the right. It becomes. Cross talk is indicated by the overlapping ratio of spots 37 and 34. Here, the parameters of d, f, D and g are equal at the light emitting point on the left side and at the light emitting point on the right side, and the average distance between both light emitting points is p. Also, although not shown in FIG. 1, the spot of parallel luminous flux emitted from the right end of the luminous point adjacent to the right of the luminous point on the right side similarly overlaps the spot 37 and crosses in detection of the luminous point 15 on the right It will be a talk. In order to detect light emission from each light emitting point well, the crosstalk should be as small as possible and at least smaller than the signal strength. Therefore, in the present invention, since crosstalk from the light emitting points on both sides of the light emitting point to be generated occurs equally, each crosstalk signal intensity ratio is suppressed to 25% or less to reduce the crosstalk of each light emitting point. The goal is to achieve detection.

FIG. 4 is a graph of the horizontal axis g and the vertical axis f showing the relationship between g and f that satisfy the crosstalk signal strength ratio of 25% or less. Here, the average effective diameter of the light emitting point is d = 0.05 mm. Here, p is a parameter and D = p. It was found that the crosstalk signal intensity ratio is 25% or less if g and f in the region above the straight line having a positive inclination passing through the origin, regardless of the value of p. The larger the parameter p, the smaller the slope, and the larger the area satisfying the condition. As a result of detailed analysis, the region that satisfies the condition is generally
f ≧ 0.95 * (d / p) * g (2)
It turned out that it can express with. Unlike in the case of the relative detection light amount of FIG. 3, the crosstalk can be suppressed to a lower value as f is larger and g is smaller. That is, for f, it was found that there is a trade-off relationship between increasing the relative detection light quantity and reducing the crosstalk.

  FIG. 5 (a) shows the relationship between g and f satisfying the condition of 50% or more in relative detected light amount and 25% or less in crosstalk signal intensity ratio by the graphs of horizontal axis g and vertical axis f. is there. FIG. 5 (b) is an enlarged view of FIG. 5 (a). Here, the average effective diameter of the light emitting point is d = 0.05 mm, and D = p. Needless to say, the area satisfying this condition is an area where the area of FIG. 3 and the area of FIG. 4 overlap. The larger the parameters D and p, the larger the area satisfying the condition. A region that satisfies this condition can generally be expressed by Equation (1) and Equation (2).

Examples of the present invention will be described below.
Example 1
FIG. 6 is a schematic view showing a device configuration example of a capillary array DNA sequencer. The analysis procedure will be described with reference to FIG. First, the sample injection end 2 of a plurality of capillaries 1 (four capillaries are shown in FIG. 6) is immersed in the cathode side buffer 4, and the sample elution end 3 is immersed in the anode side buffer 5 via the polymer block 9. The valve 14 of the pump block 9 is closed, and the polymer solution inside is pressurized by the syringe 8 connected to the pump block 9 so that the polymer solution is filled in each capillary 1 from the sample elution end 3 toward the sample injection end 2 Do. Next, the valve 14 is opened, different samples are injected from each sample injection end 2, and capillary electrophoresis is started by applying a high voltage between the cathode 6 and the anode 7 by the power supply 13. The DNA labeled with four-color fluorophores is electrophoresed from the sample injection end 2 to the sample elution end 3. The position (laser irradiation position 12) of each capillary 1 electrophoresed at a certain distance from the sample injection end 2 (laser irradiation position 12) has its coating removed and arranged on the same plane, and the laser beam 11 oscillated from the laser light source 10 After being condensed, they are introduced along the array plane from the side of the array plane, and the laser irradiation positions 12 of the respective capillaries 1 are collectively irradiated. The four-color fluorescent substance-labeled DNA is excited when passing the laser irradiation position 12 and emits fluorescence. Each emitted fluorescence is detected by the fluorescence detection device from the direction perpendicular to the arrangement plane (the direction perpendicular to the sheet of FIG. 6). The inside of the capillary constitutes a channel. Thus, the capillary array is a kind of channel array.

  FIG. 7 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting the light emission from the light emitting point array by using individual condenser lenses and a common wavelength dispersion element. FIG. 7 (a) shows a cross section perpendicular to the long axis of each capillary at the laser irradiation position, and FIG. 7 (b) shows a cross section parallel to the long axis of any one capillary. FIG. 7C shows an image detected by the two-dimensional sensor.

  As shown in FIG. 7, the four capillaries 1 with an outer diameter of 0.36 mm and an inner diameter of 0.05 mm are arranged on the same plane at an interval p of 1 mm at the laser irradiation position and narrowed to a diameter of 0.05 mm. By irradiating the beam 11 from the side of the arrangement plane, a light emitting point array in which light emitting points 15 of several n = 4 pieces and effective diameter d = 0.05 mm are arranged at an interval p = 1 mm is obtained. The four capillaries 1 constitute a capillary array, ie, a channel array. The total width of the light emitting point array is W = p * (n-1) = 3 mm. The focal position of each condenser lens 18 and each light emitting point 15 coincide with the individual condenser lens array 17 in which four condenser lenses 18 of focal length f = 2 mm and effective diameter D = 1 mm are arranged at an interval p = 1 mm In addition, the optical axis of each condenser lens 17 is set so as to be perpendicular to the array plane, and the light emission 16 from each light emitting point 15 is condensed to form a parallel luminous flux 19. Next, the parallel light beams 19 are transmitted in parallel through the common long pass filter 20 disposed parallel to the arrangement plane to cut the laser light. Subsequently, each parallel luminous flux 19 is transmitted parallel to the array plane and transmitted through one common transmission diffraction grating 21 having a grating frequency of 1000 lines / mm (grating constant 1 μm), and the major axis of each capillary 1 Disperse wavelength in the direction. The effective diameter of the transmission type diffraction grating 21 in the light emitting point array direction is DG1 = 5 mm, and the effective diameter of each capillary in the major axis direction is DG2 = 3 mm.

  At this time, as shown in FIG. 7B, the first-order diffracted lights of 500 nm, 600 nm, and 700 nm are 30.0 °, 36.9 °, and 44.4 °, respectively, with respect to the normal of the array plane. Proceed in the direction of Subsequently, an individual imaging lens array in which four imaging lenses 23 of focal length f ′ = 2 mm and effective diameter D ′ = 1 mm are arranged at an interval p ′ = 1 mm, the optical axis of each imaging lens 23 is arrayed It is inclined by 36.9 ° with respect to the normal of the plane, aligned with the optical axis of the first order diffracted light of 600 nm, and placed close to the transmission type diffraction grating 21 to equalize each wavelength dispersed parallel beam 22 Imagine. Collimated light beams 22 of 500 nm, 600 nm, and 700 nm are focused by the imaging lens 23 into focused light fluxes 24, 25 and 26, respectively. Here, the distance (optical path length) between the focusing lenses 18 and the corresponding imaging lenses 23 is set to g = 5 mm with the parallel beam 22 of 600 nm as a reference. At this time, for f = 2 mm, −0.20 * (d / D) * g + 2.8 * D = 2.75 mm, and the equation (1) is satisfied, and the relative detection light amount is 96% (> 50%) ). Further, 0.95 * (d / p) * g = 0.24 mm, and the equation (2) is satisfied, and the crosstalk signal strength ratio becomes 0.4% (<25%). Further, the sensor surface 27 of one common two-dimensional CCD is disposed parallel to the imaging lens array at a position 2 mm away from the imaging lens array, and the wavelength dispersion image 47 of each light emission 16 is detected.

  FIG. 7C shows an image 51 detected by a two-dimensional CCD, and the wavelength dispersion images 47 of the respective light emission 16 are arranged at an interval of 1 mm. Each wavelength dispersion image 47 includes imaging spots 48, 49, 50 of 500 nm, 600 nm, and 700 nm focused fluxes 24, 25, 26, respectively. Since the direction of wavelength dispersion and the direction of the light emitting point array are perpendicular, the wavelength dispersion images of each light emission are detected independently without overlapping each other. Assuming that the pixel size of the CCD is 0.05 mm square, wavelength resolution of about 20 nm / pixel can be obtained. Further, since each light emitting point 15 is imaged at the same magnification, the imaging size in the case of not being wavelength dispersed is 0.05 mm, that is, equal to the pixel size. That is, the imaging size does not reduce the wavelength resolution. When four-color detection is performed, each peak wavelength is in the range of 500 to 700 nm, and the interval is often 20 to 30 nm. Therefore, each peak wavelength has an interval of one pixel or more on the CCD and can be identified.

  By analyzing the time-dependent change of the wavelength dispersion image corresponding to each light emission point, that is, the time change of the wavelength dispersion spectrum, the time change of the fluorescence intensity of four colors is determined, and the order of base species, ie, the base sequence is determined. Since the length of the wavelength dispersion image of 500 to 700 nm is about 0.5 mm, it is sufficient that the size of the sensor surface of the two-dimensional CCD is 5 mm or more in the light emitting point array direction and 1 mm or more in the wavelength dispersion direction.

The overall size of the fluorescence detection device described above is based on the volume (250 mm ² ) of a rectangular solid defined with a width of 5 mm in the long axis direction of the capillary, 10 mm in the direction perpendicular to the array plane, and 5 mm in the light emitting point array direction. Too small. That is, as compared with the case of Patent Document 1, the overall size of the fluorescence detection device can be reduced to 1/6, 400 times. In addition, since all of the optical elements to be used are minute, significant cost reduction is possible. Furthermore, the multicolor detection sensitivity of each light emission by the present fluorescence detection device is high and uniform, the multicolor identification accuracy is high, and the crosstalk is also low. Although the number of light emitting points is n = 4 in the above embodiment, the number is not limited, and the same effect can be exhibited even if the number is increased. A dispersive prism may be used instead of the transmissive diffraction grating. In the above, the DNA sequence by electrophoresis using a plurality of capillaries and four-color fluorescence detection were targeted, but the object of the present invention is limited to any of capillary, DNA sequence, and four-color fluorescence detection Rather, it covers all cases of multicolor detection of light emission from multiple light emission points.

  In the above, as shown in FIG. 7B, the optical path of each parallel beam is inclined from the normal to the arrangement plane with wavelength dispersion. For this reason, since it is necessary to incline the two-dimensional CCD with respect to the array plane, in some cases, steric hindrance may occur with other elements. As shown in Fig. 7 (b), the sensor surface of the CCD may have a width of 1 mm or more in the wavelength dispersion direction, which is unlikely to cause steric hindrance, but the circuit board and housing of the CCD 7) can cause steric hindrance. For example, in the case of FIG. 7, when the width of the entire CCD in the wavelength dispersion direction exceeds 27 mm (assuming that the sensor surface is located at the center of the width in the wavelength dispersion direction), the arrangement planes of the CCD and capillary collide. In order to avoid such a problem, it is preferable to arrange the capillary array plane and the CCD sensor plane in parallel.

  In order to realize this, as shown in FIG. 8, the low dispersion prism 97 is disposed downstream of the transmission diffraction grating 21 so that it is refracted in the direction opposite to the traveling direction of each parallel light beam 22 transmitted through the transmission diffraction grating 21. The traveling directions of the parallel light beams 22 transmitted through the low dispersion prism 97 are perpendicular to the arrangement plane. As a material of the low dispersion prism 97, glass with small dispersion is used. For example, the glass material is SK16 (nd = 1.62, dd = 60.3), and one side of a low dispersion prism with an apex angle of 50 ° is disposed close to the transmission diffraction grating 21 in parallel with the array plane. At this time, a parallel beam with a wavelength of 600 nm enters the low dispersion prism 97 from one side at an incident angle of 36.9 °, and exits from the other side at an emission angle of 50 °, that is, in the direction perpendicular to the array plane. The optical path length between each condenser lens 18 and each corresponding imaging lens 23 is kept at g = 5 mm, and the relative detected light quantity and the crosstalk signal intensity ratio are the same as described above. Therefore, the sensor plane 27 of the two-dimensional CCD and the array plane of the capillary 1 can be made parallel while having multicolor detection performance equivalent to that of FIG. 7, and steric hindrance of the two-dimensional CCD and array plane can be avoided. Such a configuration is more effective as the fluorescence detection device is miniaturized. Although the low dispersion prism 97 is disposed downstream of the transmission diffraction grating 21 here, it may be disposed at another position. When a dispersion prism is used instead of the diffraction grating as the wavelength dispersion element, a direct view prism in which the dispersion prism and the low dispersion prism are combined may be used.

  FIG. 9 is a schematic diagram showing an example of a device configuration for detecting light emitted from the light emitting point array by a polychromatic lens, a common concave reflection type diffraction grating, and a sensor disposed perpendicularly to the optical axis of the condensing lens. It is. FIG. 9 (a) shows a cross section perpendicular to the long axis of each capillary at the laser irradiation position, and FIG. 9 (b) shows a cross section parallel to the long axis of any one capillary.

  As shown in FIG. 9, if a common concave reflection type diffraction grating 38 is used instead of the common transmission type diffraction grating, the diffraction grating doubles as an imaging lens, so that the individual imaging lens array can be omitted, and fluorescence Further miniaturization of the detection device is possible. Also in this case, the sensor plane 27 of the two-dimensional sensor and the array plane of the capillaries can be arranged in parallel to avoid steric hindrance caused by the miniaturization. The direction of wavelength dispersion is made to coincide with the long axis direction of the capillary, as described above. Each condenser lens 18 has the same focal length f = 2 mm, effective diameter D = 1 mm, interval p = 1 mm as above, but the distance between each condenser lens 18 and the concave reflection type diffraction grating 38 is g = 2 mm, The focal length of the concave reflection type diffraction grating 38 is f ′ = 4 mm. At this time, the relative detection light amount is improved to 98%, and the crosstalk signal intensity ratio is reduced to 0.1%. However, as it is, since the imaging lens (concave reflection type diffraction grating 38) is made common, each parallel luminous flux 19 is imaged at the same position, and the wavelength dispersive images of each light emission 16 coincide on the two-dimensional sensor Therefore, each light emission 16 can not be detected in multiple colors independently. Here, in order to solve this problem, the imaging positions of the respective parallel light beams 19 are shifted by mutually shifting the optical axes of the focusing lenses 18 of the individual focusing lens arrays 17 from parallel.

For example, as shown in FIG. 9A, the angle formed by the optical axis of each condenser lens 18 indicated by the alternate long and short dash line and the normal to the array plane indicated by the solid line is θ ₁ = 3 °, θ ₂ = 1 ° , Θ ₃ = −1 °, θ ₄ = −3 °, so that each parallel luminous flux 19 is spread radially. However, the focal positions of the condenser lenses 18 are arranged so as not to shift from the light emitting points 15. At this time, the imaging positions of each light emission are separated from each other by a distance of 0.14 mm, and this distance is larger than the imaging size of each light emission 0.1 mm (because the image magnification is doubled). It is possible to detect in multiple colors.

  As shown in FIG. 7, when the laser beam 11 is irradiated from the side of the array plane of the plurality of capillaries 1, the laser irradiation intensity of each capillary is the incident side of the laser beam 11 by the laser reflection at the interface of each capillary. It gradually decreases from the right side of 7 (a) to the emission side (left side of FIG. 7 (a)). Therefore, even if the fluorescence detection devices after the condenser lens 18 have the same efficiency for each light emitting point 15, the obtained fluorescence detection intensity or sensitivity is determined by the capillary 1 after the laser beam irradiation (see FIG. The left side of a) may become lower. In order to eliminate such nonuniformity, it is effective to change the condensing efficiency of each condensing lens 18 for each light emitting point 15. For example, it is effective to make the effective diameter of the condensing lens 18 on the incident side of the laser beam 11 smaller and make the effective diameter of the condensing lens 18 on the outgoing side of the laser beam 11 larger.

Example 2
FIG. 10 is a schematic view showing an apparatus configuration example for detecting light emission from a light emission point array by using individual condenser lenses, a common dichroic mirror set, and a sensor. FIG. 10 (a) shows a cross section perpendicular to the long axis of each capillary at the laser irradiation position, and FIG. 10 (b) shows a cross section parallel to the long axis of any one capillary. Further, FIG. 10C shows an image detected by the two-dimensional sensor.

  As shown in FIG. 10, the laser irradiation positions of four capillaries 1 having an outer diameter of 0.36 mm and an inner diameter of 0.05 mm are arranged on the same plane at an interval p of 0.5 mm and the diameter is narrowed to 0.05 mm. By irradiating the beam 11 from the side of the arrangement plane, a light emitting point array in which light emitting points 15 of several n = 4 and effective diameter d = 0.05 mm are arranged at an interval p = 0.5 mm is obtained. Here, the wavelength of the laser beam 11 is 505 nm, and the fluorescence (emission maximum wavelength) of four colors is A fluorescence (540 nm), B fluorescence (570 nm), C fluorescence (600 nm), and D fluorescence (630 nm). The total width of the light emitting point array is W = p * (n-1) = 1.5 mm. The individual condenser lens array 17 in which four condenser lenses 18 of focal length f = 1 mm and effective diameter D = 0.4 mm are arranged at an interval p = 0.5 mm, the focal position of each condenser lens 18 and each light emission The light beams from the respective light emitting points 15 are condensed to form parallel light beams 19 so that the points 15 coincide with one another and the optical axes of the respective condensing lenses 18 are perpendicular to the arrangement plane.

  Next, each collimated light beam 19 is incident in parallel on a set of common dichroic mirrors arranged in series in the optical axis direction. The dichroic mirror set is composed of five elements of a long pass filter 56, an A dichroic mirror 39, a B dichroic mirror 41, a C dichroic mirror 43, and a D dichroic mirror 45, and each element is one each. It is used in common and in parallel for each light emitting point. The long pass filters 56 are arranged in parallel to the array plane at a distance of 0.5 mm from each condenser lens 18. The dichroic mirrors 39, 41, 43, 45 are disposed at intervals of 1 mm in parallel with the long axis of the capillary, and are disposed such that the normals are inclined 45 ° with respect to the array plane. Further, the center of the A dichroic mirror 39 is disposed at a distance of 1 mm from each condenser lens (a distance of 0.5 mm from the long pass filter 56). As for the size of each element, the effective diameter in the arrangement direction of the light emitting point array is DM1 = 3 mm, and the effective diameter in the orthogonal direction is DM2 = 1.4 mm (only DM2 = 1 mm for the long pass filter).

  First, each collimated light beam 19 is vertically incident in parallel to the long pass filter 56 to cut light of 520 nm or less, and in particular to cut 505 nm which is the wavelength of the laser beam. The respective parallel light beams transmitted through the long pass filter 56 are then made to be incident in parallel at 45 ° to the A dichroic mirror 39 to transmit light of 530 to 550 nm and reflect light of 560 nm or more. Each parallel light flux of 530 to 550 nm, which is the transmitted light of the A dichroic mirror 39, is called an A parallel light flux 40 and is mainly used for detection of A fluorescence (maximum emission wavelength 540 nm). Next, parallel light fluxes reflected by the A dichroic mirror 39 are made to enter the B dichroic mirror 41 in parallel at 45 °, and light of 560 to 580 nm is reflected and light of 590 nm or more is transmitted. Each parallel light flux of 560 to 580 nm which is the reflected light of the B dichroic mirror 41 is called a B parallel light flux 42, and is mainly used for detection of B fluorescence (maximum emission wavelength 570 nm).

  Next, parallel light fluxes transmitted by the B dichroic mirror 41 are made to enter the C dichroic mirror 43 in parallel at 45 °, and light of 590 to 610 nm is reflected and light of 620 nm or more is transmitted. Each parallel light beam of 590 to 610 nm which is the reflected light of the C dichroic mirror 43 is called a C parallel light beam 44 and is mainly used for detection of C fluorescence (maximum emission wavelength 600 nm). Each parallel light flux which is the transmitted light of the C dichroic mirror 43 is then made to be incident in parallel at 45 ° to the D dichroic mirror 45 to reflect light of 620 to 640 nm and transmit light of 650 nm or more (shown in FIG. 10) ). A parallel light flux of 620 to 640 nm, which is the reflected light of the D dichroic mirror 45, is called a D parallel light flux 46 and is mainly used for detection of D fluorescence (maximum emission wavelength: 630 nm). All four parallel light beams 40, 42, 44, 46 corresponding to the four light emitting points travel in the direction perpendicular to the arrangement plane. The D dichroic mirror 45 may be replaced by a total reflection mirror.

  Subsequently, the sensor surface 27 of the common two-dimensional CCD is disposed parallel to the array plane at a distance of 2 mm from each condenser lens (a distance of 1 mm from the center of each dichroic mirror 39, 41, 43, 45), The four sets of parallel light beams 40, 42, 44 and 46 are incident in parallel on the sensor surface 27 without imaging. On the image 51 captured by the two-dimensional CCD shown in FIG. 10C, four spots 52, 53, 54, and 55 corresponding to the parallel light beams 40, 42, 44, and 46, respectively, have a total of 16 spots. It is formed. The spots are arranged in a grid at a diameter of 0.4 mm, at an interval of 0.5 mm in the light emitting point array direction, and at an interval of 1 mm in the long axis direction of the capillary, and are independently detected. Therefore, the size of the sensor surface 27 of the two-dimensional CCD should be 3 mm or more in the light emitting point array direction and 5 mm or more in the capillary long axis direction. At this time, based on the D-parallel beam 46 with the longest optical path length, the distance between each condenser lens 18 and the sensor surface 27 is g = 5 mm. For f = 1 mm, −0.20 * (d / D) * g + 2.8 * D = 1 mm, and the equation (1) is satisfied, and the relative detection light amount is 51% (> 50%). In addition, 0.95 * (d / p) * g = 0.48 mm, and the equation (2) is satisfied, and the crosstalk signal strength ratio becomes 0.1% (<25%). By analyzing the time change of the intensity of the four spots corresponding to each light emission point, the time change of the fluorescence intensity of four colors is determined, and the order of the base species, that is, the base sequence is determined.

The overall size of the fluorescence detection device described above is based on the volume (75 mm ² ) of a cuboid whose width in the long axis direction of the capillary is 5 mm, width in the direction perpendicular to the array plane is 5 mm, and width in the light emitting point array direction is 3 mm. Too small. That is, as compared with the case of Patent Document 1, the entire size of the fluorescence detection device can be reduced to 1 / 21,000 times. In addition, since all of the optical elements to be used are minute, significant cost reduction is possible. Furthermore, the multicolor detection sensitivity of each light emission by the present fluorescence detection device is high and uniform, the multicolor identification accuracy is high, and the crosstalk is also low. Although the number of light emitting points is n = 4 in the above embodiment, the number is not limited, and the same effect can be exhibited even if the number is increased. Another effect of performing multicolor detection using a dichroic mirror set is that the amount of effectively detected light is large as compared with the case of the diffraction grating used in Patent Document 1 and Example 1. When a diffraction grating is used, the diffraction efficiency that can be used for wavelength dispersion is about 50%, but when a dichroic mirror set is used, there is almost no loss. An amount of light can be obtained.

  FIG. 11 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting light emission from the light emission point array with an individual condensing lens, a common dichroic mirror set, and a sensor as polychromatic in the same manner as wavelength dispersion.

  FIG. 11 is a development of the above-mentioned common dichroic mirror set. The dichroic mirror set is composed of nine elements of a long pass filter 56 and dichroic mirrors 57, 59, 61, 63, 65, 67, 69, 71 arranged in order in the optical axis direction. The dichroic mirror 57 transmits parallel light beams 58 of 520 to 540 nm and reflects light of 540 nm or more. The dichroic mirror 59 reflects the parallel beam 60 of 540 to 560 nm and transmits light of 560 nm or more. The dichroic mirror 61 reflects the parallel light beam 62 of 560 to 580 nm and transmits light of 580 nm or more. The dichroic mirror 63 reflects the parallel beam 64 of 580 to 600 nm and transmits light of 600 nm or more. The dichroic mirror 65 reflects the parallel light beam 66 of 600 to 620 nm and transmits light of 620 nm or more. The dichroic mirror 67 reflects the parallel luminous flux 68 of 620 to 640 nm and transmits light of 640 nm or more. The dichroic mirror 69 reflects the parallel beam 70 of 640 to 660 nm and transmits light of 660 nm or more. The dichroic mirror 71 reflects the parallel light beam 72 of 660 to 680 nm and transmits light of 680 nm or more.

  As described above, the light emission from one light emitting point 15 forms eight spots on the sensor surface 27, and these intensities give a 20 nm wide emission spectrum in the range of 520 to 680 nm. With such a configuration, it is not necessary to redesign the dichroic mirror according to the type of phosphor used, and it becomes possible to detect any fluorescence in the range of 520 to 680 nm with high sensitivity and high accuracy. . It goes without saying that the number of spots for dividing each light emission and the wavelength width for dividing are not limited to the above embodiment, and can be set arbitrarily.

[Example 3]
FIG. 12 is a schematic cross-sectional view showing an example of the apparatus configuration for detecting the light emission from the light emission point array by using individual condenser lenses and color sensors. FIG. 12 (a) shows a cross section perpendicular to the long axis of each capillary at the laser irradiation position, and FIG. 10 (b) shows an image detected by the two-dimensional sensor. The present embodiment is an example in which a color sensor surface 73 is used for a two-dimensional sensor, for example, a two-dimensional CCD.

  Each light emitting point 15 to the long pass filter 56 have the same configuration as that of FIG. 10 of the second embodiment. As shown in FIG. 12A, each parallel beam transmitted through the long pass filter 56 is directly incident on the color sensor surface 73 of the two-dimensional CCD. The distance between each condenser lens 18 and the color sensor surface 73 is g = 1 mm. At this time, for f = 1 mm, −0.20 * (d / D) * g + 2.8 * D = 1.1 mm, equation (1) is satisfied, and the relative detection light amount is 61% (> 50%) ). Further, 0.95 * (d / p) * g = 0.10 mm, and the equation (2) is satisfied, and the crosstalk signal strength ratio becomes 0.0% (<25%).

  As shown in FIG. 12B, spots 75 of parallel light beams are formed on the image 74 of the color sensor surface 73. The spots 75 are arranged at a diameter D of 0.4 mm and at an interval of 0.5 mm in the light emitting point array direction, and each of the spots 75 is independently detected. As schematically shown in the enlarged view of FIG. 12 (b), the color sensor surface 73 mainly comprises an A pixel 76 for detecting A fluorescence (maximum emission wavelength 540 nm) and mainly B fluorescence (maximum emission wavelength 570 nm). The B pixel 77 to detect, the C pixel 78 mainly to detect C fluorescence (maximum emission wavelength 600 nm), and the D pixel 79 mainly to detect D fluorescence (maximum emission wavelength 630 nm) It is arranged and arranged. The size of each pixel 76, 77, 78, 79 is S = 0.05 mm, and S <D is satisfied. At this time, each spot 75 is detected by about 80 pixels, and is detected by about 20 pixels per one type of pixel. As described above, the multicolor detection of the light emission from each light emitting point 15 can be performed with high accuracy by the large number of pixels detecting each spot 75 for each type of pixel. For example, there is no problem even if the relative position of each type of pixel and spot changes. Alternatively, even if the light intensity distribution in the spot is nonuniform, each color can be detected uniformly.

The overall size of the fluorescence detection device described above is based on the volume (18 mm ² ) of a cuboid whose width in the long axis direction of the capillary is 3 mm, width in the direction perpendicular to the array plane is 2 mm, and width in the light emitting point array direction is 3 mm. Too small. That is, as compared with the case of Patent Document 1, the entire size of the fluorescence detection device can be reduced to 1 / 89,000 times. In addition, since all of the optical elements to be used are minute, significant cost reduction is possible. Furthermore, the multicolor detection sensitivity of each light emission by the present fluorescence detection device is high and uniform, the multicolor identification accuracy is high, and the crosstalk is also low. Although the number of light emitting points is n = 4 in the above embodiment, the number is not limited, and the same effect can be exhibited even if the number is increased. However, when the color sensor as described above is used, the utilization efficiency of the amount of light incident on the sensor surface is reduced to 1⁄4. As compared with the case of the diffraction efficiency of about 50% of Example 1 using the diffraction grating, the effective efficiency is about half. However, the device configuration is very simple and further miniaturization is possible.

  In order to improve the utilization efficiency of the light quantity incident on the sensor surface, instead of using a color sensor in which elements for detecting each color are arranged in parallel with the sensor surface as shown in FIG. Is effective. In this case, it is not necessary to satisfy S <D.

Example 4
One of the mounting problems of the present invention is how to accurately align the light emitting points with the condenser lenses with ease. The present embodiment shows means for realizing this for a plurality of capillaries.

  FIG. 13 is a schematic cross-sectional view showing a configuration example of a device in which a plurality of capillaries, a V-groove array in which the plurality of capillaries are arranged, and an individual condensing lens array are integrated. 13 (a) shows a cross section perpendicular to the long axis of each capillary at the laser irradiation position, and FIG. 13 (b) shows a cross section perpendicular to the long axis of each capillary at a location not at the laser irradiation position. c) shows a cross section parallel to the long axis of any one capillary. 13 (a) corresponds to the cross section AA in FIG. 13 (c), and FIG. 13 (b) corresponds to the cross section BB in FIG. 13 (c).

  The device shown in FIG. 13 includes a capillary array consisting of a plurality of capillaries 1 and a sub device 80. In the sub device 80, a V groove array in which a plurality of V grooves 81 are arranged at an interval p and a condensing lens array in which a plurality of condensing lenses 18 are arranged at an interval p are formed in the same device. The central axes of the grooves 81 and the focusing lenses 18 are made to coincide with each other. The plurality of capillaries 1 can be arranged on the same plane at a predetermined interval p simply by pressing the plurality of capillaries 1 against the V groove 81 respectively. In addition, the relative positions of the V grooves 81 and the focusing lenses 18 in the sub device 80 such that the light emitting points 15 at which the laser beams 11 of the capillaries 1 are irradiated coincide with the focal points of the focusing lenses 18. Adjust the Thus, the light emitted from the light emitting point 15 is converted into a parallel light beam 19 by the condensing lens 18.

  As shown in FIG. 13A, the condenser lens 18 of the sub device 80 exists in the cross section of the capillary 1 at the light emitting point 15, and the V groove 81 does not exist. On the other hand, as shown in FIG. 13B, in the cross section of the capillary 1 on both sides of the light emitting point 15, the condenser lens 18 of the sub device 80 does not exist, and the V groove 81 exists. FIG. 13C shows a cross section of the capillary 1 in the long axis direction. A condenser lens 18 exists at the center of the sub device 80, and V grooves 81 exist at both sides thereof. This is a device to prevent the V groove 81 from disturbing the detection of the light emission from the light emitting point 15 while realizing highly accurate alignment of the capillary 1 by the V groove 81. If the sub device 80 as described above is prepared in advance, high precision alignment between each light emitting point 15 and each condenser lens 18 can be easily performed simply by pressing the plurality of capillaries 1 onto each V groove 81 respectively. It becomes possible. The present embodiment can be combined with any of the configurations of the first to third embodiments. The sub device 80 in which the V-groove array and the condenser lens array are integrated can be integrally molded by a processing method such as injection molding or imprint, and mass production is also possible at low cost. Of course, the sub-device 80 may be completed by separately fabricating the V-groove 81 and the condenser lens 18 and then combining them.

  The sub device is also effective when there is no V-groove array. For example, the surface on the capillary array side of the sub device may be a plane instead of the V-groove array. Although it is necessary to adjust the arrangement interval of the plurality of capillaries by another means, the distance between each capillary and each condensing lens, that is, each light emitting point and each condensing lens, is required by pressing each capillary against the above-mentioned plane of the subdevice. It is possible to control the distance of Alternatively, even if it is not the V groove, a structure for controlling the position of the capillary may be provided in the sub device.

  The focal length f1 of each condenser lens 18 in the light emitting point array direction and the focal length f2 of the capillary in the long axis direction are f1 = f2 in the above embodiments, but f1 ≠ f2 is also effective. It is. Since the capillary 1 has a cylindrical shape, it has a lens action in the light emitting point array direction, but has no lens action in the long axis direction. Therefore, in order to efficiently condense the light emitted from the light emitting point 15 by the condenser lens 18, it is effective to cancel the difference due to the direction of the lens action of the above-mentioned capillary. Just do it. This can be easily realized by making each condenser lens 18 aspheric. In addition, by forming each condenser lens 18 as a Fresnel lens, it is possible to reduce the thickness of the lens and further miniaturize the fluorescence detection device. The use of a Fresnel lens is, of course, also effective when f1 = f2.

  FIG. 14 is a schematic cross-sectional view showing a configuration example of a device in which individual condenser lenses are adhered to a plurality of capillaries. FIG. 14 (a) shows a cross section perpendicular to the long axis of each capillary at the laser irradiation position, and FIG. 14 (c) shows a cross section parallel to the long axis of any one capillary. Here, another method is described in which the alignment between each light emitting point and each condenser lens is performed accurately with ease.

  The individual condensing lenses 18 are adhered to the respective capillaries 1 so that the focal points of the respective condensing lenses 18 and the light emitting points 15 of the respective capillaries 1 coincide with each other. In FIG. 14, although the spherical condensing lens 18 is used, of course, the condensing lens of another shape may be used. The adhesion of each condenser lens 18 to each capillary 1 is preferably performed after the alignment of the plurality of capillaries on the same plane is completed. This has the effect of avoiding that the plurality of focusing lenses 18 are not aligned on the same plane or that the optical axes of the plurality of focusing lenses 18 are not parallel to each other in the step of arranging the plurality of capillaries 1 . In addition, by supplying the capillary array in a state in which the above adhesion has been completed to the user, the relative positions of each light emitting point 15 and each condensing lens 18 in the transportation step of the capillary array, the installation step to the fluorescence detection device, etc. It is possible to prevent deviation from the predetermined position.

[Example 5]
FIG. 15 is a schematic cross-sectional view showing a configuration example of a device in which a microchip having a multi-channel and an individual condensing lens array are integrated. In this embodiment, not a plurality of capillaries but a plurality of channels 82 provided in the microchip 86, that is, a channel array is targeted.

  The microchip 86 in the illustrated example is manufactured by bonding a channel substrate 83 having a plurality of rectangular four grooves formed on the surface thereof and a planar substrate 84 having a flat surface, with the surfaces thereof facing each other. Ru. The boundary between the channel substrate 83 and the flat substrate 84 is called a bonding surface 85. The four grooves described above are separated by a bonding surface 85 to form four channels 82. These channels 82 have a diameter of 0.05 mm and are arranged on the same plane at a spacing p = 0.5 mm. In this embodiment, the array plane of the plurality of channels is simply referred to as an array plane. By irradiating the laser beam 11 narrowed to a diameter of 0.05 mm from the side of the array plane, a light emitting point where several n = 4 light emitting points 15 of effective diameter d = 0.05 mm are arrayed at an interval p = 0.5 mm Get an array. The total width of the light emitting point array is W = p * (n-1) = 1.5 mm. Further, in the present embodiment, four individual condenser lenses 18 are formed on the back surface of the channel substrate 83 opposite to the surface on which the grooves are formed. These condenser lenses 18 are arranged parallel to the arrangement plane at an interval p = 0.5 mm so that each optical axis is perpendicular to the arrangement plane, and each focal point coincides with each light emission point.

  If the channel substrate 83 is manufactured by injection molding or imprinting, it is possible to process it at low cost while aligning the relative positions of the grooves on the front surface and the condensing lens 18 on the rear surface precisely as described above. Here, the focal length f of the condenser lens 18 is 1 mm, and the effective diameter D is 0.4 mm. The light emitted from each light emitting point 15 is condensed by the condensing lens 18 to form a parallel luminous flux 19. After that, it can be combined with the fluorescence detection device of any of the embodiments described above. The DNA sequence by electrophoresis may be performed using each channel 82 in the same manner as the above embodiment, or may be applied to other applications. In any case, light emission from the four light emission points can be detected with low color crosstalk and high sensitivity using the fluorescence detection device which is much smaller than that of the prior art.

  Next, a more specific embodiment using the microchip 86 will be described.

  FIG. 16 shows an apparatus configuration example for detecting multicolor light emission from a device in which a microchip having multichannels and an individual condensing lens array are integrated and light emission point array by individual LED illumination using a common dichroic mirror set and a sensor It is a cross-sectional schematic diagram shown. 16 (a) is a schematic top view of the microchip, FIG. 16 (b) is a schematic cross-sectional view of one arbitrary channel viewed from the side, and FIG. 16 (c) is a liquid that is fluorescently labeled and flows inside the channel It is an explanatory view showing a relation of a drop and a luminescence point.

  As shown in the schematic top view of FIG. 16A, in the microchip 86 of the illustrated example, ten channels 82 with a diameter of 0.1 mm and a length of 50 mm are parallel and in the same plane at an interval p = 2 mm. Arranged on top. At both ends of each channel 82, a flow inlet 87 and a flow outlet 88 are formed. A light emitting point 15 is located at the center of each channel 82.

  As shown in FIG. 16 (b), unlike the previous embodiments, in this embodiment, LED light is used as excitation light instead of a laser beam, and an optical system for incident fluorescence detection is adopted. Also, the LED light sources 90 are individually prepared for each channel 82. The LED light having a center wavelength of 505 nm oscillated from the individual LED light source 90 is made into an LED parallel luminous flux 92 by the individual collimating lens 91 with a focal distance of 5 mm, and then incident in parallel to the common LED dichroic mirror 89 at an incident angle of 45 °. It is reflected and travels perpendicularly to the array plane. The center of the LED dichroic mirror 89 is disposed at a distance of 1 mm from each condenser lens 18. Next, each LED parallel luminous flux 92 is condensed at the position of each light emitting point 15 by an individual condensing lens 18 of focal length f = 1 mm and effective diameter D = 1 mm. At this time, since the condensed size of the LED light is 0.05 mm in diameter, the size of the light emitting point is also d = 0.05 mm, and the diameter of the channel 82 can be smaller than 0.1 mm. This is advantageous in reducing crosstalk.

  The light emitted from each light emitting point 15 is collimated by the same individual condenser lens 18 into parallel light beams 19 and incident in parallel to the common LED dichroic mirror 89 at an incident angle of 45 °, and the LED light is respectively reflected by the LED dichroic mirror 89 The light is reflected and travels in the direction of the LED light source 90, and the fluorescence is transmitted through the LED dichroic mirror 89, respectively. The following elements of A dichroic mirror 39, B dichroic mirror 41, C dichroic mirror 43, and D dichroic mirror 45 are used in common and in parallel for each light emitting point, and A fluorescence, B fluorescence, C fluorescence and D fluorescence are used. The point to be detected is the same as that of the second embodiment. The difference from the second embodiment is that the distance between the center of the A dichroic mirror 39 and each condenser lens 18 is 2 mm, and the effective diameter of the light emitting point array of each dichroic mirror 39, 41, 43, 45 is DM1. = 25 mm. At this time, the distance between each condenser lens 18 and the sensor surface 27 is g = 6 mm. For f = 1 mm, -0.20 * (d / D) * g + 2.8 * D = 2.74 mm, equation (1) is satisfied, and the relative detection light quantity is 362% (> 50%) . Further, 0.95 * (d / p) * g = 0.14 mm, and the equation (2) is satisfied, and the crosstalk signal strength ratio becomes 0.0% (<25%).

  In this embodiment, the microchip 86 and the fluorescence detection device described above are used for the measurement of digital PCR. In digital PCR, a large number of droplets (emulsion) are formed in oil, and each droplet contains zero or one target DNA molecule. In this state, PCR is performed to emit fluorescence when the target is present and amplified. The number of molecules of the target present in the original sample is quantified with high accuracy by examining the presence or absence of fluorescence of each droplet. Furthermore, by performing four-color fluorescence detection, it is possible to perform digital PCR independently for four types of targets. One of the problems of digital PCR is the improvement of throughput, and high throughput multicolor detection of many droplets is important. FIG. 16 (c) shows the flow of the light inside the channel 82 along with the oil, in which four types of droplets 93, 94, 95, 96 labeled with A fluorescence, B fluorescence, C fluorescence and D fluorescence flow. It shows a configuration in which each fluorescence is emitted upon excitation when crossing 15. Conventionally, digital PCR was measured by one-color detection using one channel, but in this example, digital PCR can be measured by four-color detection using ten channels, so the throughput It will be 40 times. Moreover, the microchip 86 and the fluorescence detection device are very compact and can be manufactured at low cost.

  The present invention is not limited to the embodiments described above, but includes various modifications. For example, the embodiments described above are described in detail in order to explain the present invention in an easy-to-understand manner, and are not necessarily limited to those having all the configurations described. Also, part of the configuration of one embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of one embodiment. In addition, with respect to a part of the configuration of each embodiment, it is possible to add, delete, and replace other configurations.

Reference Signs List 1 capillary 10 laser light source 11 laser beam 12 laser irradiation position 15 light emitting point 17 condensing lens array 18 condensing lens 20 long pass filter 21 transmission type diffraction grating 23 imaging lens 27 sensor surface 28 sensor area 38 concave reflection type diffraction grating 39 A Dichroic mirror 41 B dichroic mirror 43 C dichroic mirror 45 D dichroic mirror 47 wavelength dispersion image 56 long pass filter 73 color sensor surface 80 sub-device 81 V groove 82 channel 83 channel substrate 84 planar substrate 86 microchip 90 LED light source 91 collimate lens 97 low Dispersing prism

Claims (22)

Let m and n be arbitrary integers of 2 or more, respectively
a condenser lens array in which m condenser lenses are arranged to individually condense the light emission from each light emission point of the light emission point array in which the m luminous points are arrayed into m luminous fluxes;
A dichroic mirror array in which n dichroic mirrors are arranged substantially in parallel, including at least a first dichroic mirror and a second dichroic mirror;
At least one sensor,
At least a portion of the m beams is incident in parallel to the first dichroic mirror, and the first dichroic mirror transmits at least a portion of the m beams to the m first transmitted beams. And m first reflected beams,
At least a portion of the m first reflected beams are respectively incident in parallel to the second dichroic mirror, and the second dichroic mirror is at least a portion of the m first reflected beams. Convert to at least m second reflected beams,
The multicolor detection device according to claim 1, wherein at least a part of the m first transmitted light beams and at least a part of the m second reflected light beams are incident on the sensor in parallel without being collected again. In the multicolor detection device according to claim 1,
The multicolor detection device, wherein the direction of the optical axis of the m condenser lenses and the sensor surface of the sensor are substantially perpendicular. In the multicolor detection device according to claim 1 or 2,
A multicolor detection device in which the direction of the optical axis of the m condenser lenses, the direction in which the m condenser lenses are arranged, and the direction in which the n dichroic mirrors are arranged are substantially perpendicular. In the multicolor detection device according to claim 1,
The average of the effective diameters of the m light emitting points is d, the average of the focal distances of the m condenser lenses is f, the average of the effective diameters of the m condenser lenses is D, the m second Let g be the average of the optical path length between the m condenser lenses and the sensor of the reflected light flux of
f ≦ −0.20 * (d / D) * g + 2.8 * D
Satisfy multi-color detection device. In the multicolor detection device according to claim 4,
When the average of the arrangement interval of the m light emitting points is p,
f ≧ 0.95 * (d / p) * g
Satisfy multi-color detection device. In the multicolor detection device according to claim 1,
The average of the effective diameters of the m light emitting points is d, the average of the focal lengths of the m condenser lenses is f, the average of the arrangement intervals of the m light emitting points is p, the m second Assuming that the average of the optical path length between the m condenser lenses and the sensor of the reflected light flux is g,
f ≧ 0.95 * (d / p) * g
Satisfy multi-color detection device. In the multicolor detection device according to claim 1,
The average of the arrangement interval of the m light emitting points is p, the average of the effective diameters of the m condenser lenses is D, and the effective diameter of the m light emitting points of the n dichroic mirrors is DM1 When an effective diameter in a direction orthogonal to the arrangement direction of the m light emitting points of the n dichroic mirrors is DM2,
p * (m-1) + D ≦ DM1, and √2 * D ≦ DM2
Satisfy multi-color detection device. In the multicolor detection device according to claim 1,
The multicolor detection device in which the optical axes of the m condenser lenses are not parallel to one another. In the multicolor detection device according to claim 1,
Further comprising a third dichroic mirror not belonging to the dichroic mirror array;
m illumination beams are respectively incident on the third dichroic mirror in parallel, and the third dichroic mirror converts at least a portion of the m illumination beams into at least m third reflection beams. The
The m third reflected beams are individually condensed by the m condenser lenses, and the m light emitting points are individually irradiated,
Multicolor detection in which the m luminous fluxes are luminous fluxes in which light emissions from the m luminous points are individually condensed by the m condensing lenses and transmitted in parallel by the third dichroic mirror apparatus. Let m and n be arbitrary integers of 2 or more, respectively
a condenser lens array in which m condenser lenses are arranged to individually condense the light emission from each light emission point of the light emission point array in which the m luminous points are arrayed into m luminous fluxes;
A dichroic mirror array in which n dichroic mirrors are arranged substantially in parallel, including at least a first dichroic mirror and a second dichroic mirror;
At least one sensor,
At least a portion of the m beams is incident in parallel to the first dichroic mirror, and the first dichroic mirror transmits at least a portion of the m beams to the m first transmitted beams. And m first reflected beams,
At least a portion of the m first transmitted beams are respectively incident in parallel to the second dichroic mirror, and the second dichroic mirror transmits at least a portion of the m first transmitted beams. Convert to at least m second reflected beams,
The multicolor detection device according to claim 1, wherein at least a part of the m first reflected light beams and at least a part of the m second reflected light beams are incident on the sensor in parallel without being collected again. In the multicolor detection device according to claim 10,
The multicolor detection device, wherein the direction of the optical axis of the m condenser lenses and the sensor surface of the sensor are substantially perpendicular. In the multicolor detection device according to claim 10 or 11,
A multicolor detection device in which the direction of the optical axis of the m condenser lenses, the direction in which the m condenser lenses are arranged, and the direction in which the n dichroic mirrors are arranged are substantially perpendicular. In the multicolor detection device according to claim 10,
The average of the effective diameters of the m light emitting points is d, the average of the focal distances of the m condenser lenses is f, the average of the effective diameters of the m condenser lenses is D, the m second Let g be the average of the optical path length between the m condenser lenses and the sensor of the reflected light flux of
f ≦ −0.20 * (d / D) * g + 2.8 * D
Satisfy multi-color detection device. In the multicolor detection device according to claim 13,
When the average of the arrangement interval of the m light emitting points is p,
f ≧ 0.95 * (d / p) * g
Satisfy multi-color detection device. In the multicolor detection device according to claim 10,
The average of the effective diameters of the m light emitting points is d, the average of the arrangement interval of the m light emitting points is p, the average of the focal distances of the m condensing lenses is f, and the m second Assuming that the average of the optical path length between the m condenser lenses and the sensor of the reflected light flux is g,
f ≧ 0.95 * (d / p) * g
Satisfy multi-color detection device. In the multicolor detection device according to claim 10,
The average of the arrangement interval of the m light emitting points is p, the average of the effective diameters of the m condenser lenses is D, and the effective diameter of the m light emitting points of the n dichroic mirrors is DM1 When an effective diameter in a direction orthogonal to the arrangement direction of the m light emitting points of the n dichroic mirrors is DM2,
p * (m-1) + D ≦ DM1, and √2 * D ≦ DM2
Satisfy multi-color detection device. In the multicolor detection device according to claim 10,
The multicolor detection device in which the optical axes of the m condenser lenses are not parallel to one another. In the multicolor detection device according to claim 10,
Further comprising a third dichroic mirror not belonging to the dichroic mirror array;
m illumination beams are respectively incident on the third dichroic mirror in parallel, and the third dichroic mirror converts at least a portion of the m illumination beams into at least m third reflection beams. The
The m third reflected beams are individually condensed by the m condenser lenses, and the m light emitting points are individually irradiated,
Multicolor detection in which the m luminous fluxes are luminous fluxes in which light emissions from the m luminous points are individually condensed by the m condensing lenses and transmitted in parallel by the third dichroic mirror apparatus. Let m be any integer greater than or equal to 2,
a condenser lens array in which m condenser lenses are arranged to individually condense the light emission from each light emission point of the light emission point array in which the m luminous points are arrayed into m luminous fluxes;
At least one sensor,
At least a part of the m luminous fluxes are respectively incident on the sensor in parallel;
Each of the m light emitting points has a finite size,
The average of the effective diameters of the m light emitting points is d, the average of the focal lengths of the m condenser lenses is f, the average of the effective diameters of the m condenser lenses is D, and the m luminous fluxes , Where g is the average of the optical path length between the m condenser lenses and the sensor,
f ≦ −0.20 * (d / D) * g + 2.8 * D
Satisfy multi-color detection device. Let m be any integer greater than or equal to 2,
a capillary array in which at least a portion of m capillaries are arranged on the same plane;
A condenser lens array in which m condenser lenses arranged to individually condense light emitted from the m capillaries on the same plane individually to form m luminous fluxes;
At least one sensor,
At least a portion of the m luminous fluxes are respectively incident on the sensor in parallel without refocusing,
The multicolor detection device in which the same plane and the sensor surface of the sensor are substantially parallel to each other. The multicolor detection device according to claim 20,
The multicolor detection device, wherein the sensor is configured by arranging a plurality of kinds of pixels having different spectral sensitivity characteristics in a two-dimensional manner. In the multicolor detection device according to claim 21,
Let S be the average pixel size of the sensor,
S <D
Satisfy multi-color detection device.

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