DE112016007594B3 - LIGHT EMITTING DETECTION DEVICE - Google Patents

Abstract

A light emission detection device comprising: a condenser lens array (8) in which M condenser lenses (2) are arranged for respectively converging light emitted from a light emitting point array (7) in which M light emitting points (1) are arranged to generate M light beams where M ≥ 2, an optical element (43) which deflects the M light beams to generate M deflected light beams, and at least one sensor (11) on which the M light beams are incident in parallel, where, if an average value of the effective Is the diameter of the M light emitting points, f is an average value of the focal lengths of the M condenser lenses, p is an average value of the intermediate distances of the M condenser lenses, and g is an average value of the maximum optical path lengths between the M condenser lenses and an end face of the optical element on which the M light rays occur, is, d, f, p and g satisfy the following relation: f≥1 / ((2 * p) / (1.27 * d) +1) * g.

Description

Technical area

The present invention relates to a detection device that a plurality of channels, which are provided in a plurality of capillaries or in a microchip, irradiated with light such as from a laser beam or a lamp and fluorescence, phosphorescence, scattered light, transmitted light, etc., from a substance located in the capillary or in the channel is emitted, detects with high sensitivity.

State of the art

A DNA sequencer with a capillary arrangement (capillary array) that decodes base sequences of different DNA samples in single capillaries together by using the large number of capillaries (glass capillaries with an outer diameter of 100 µm to 400 µm and an inner diameter of 25 µm to 100 µm), which are filled with a separating medium and carry out an electrophoretic analysis in parallel processing, is widespread. This mechanism is described next. A polyimide coating film is formed on an outer surface of a commercially available capillary in order to obtain flexibility. Certain portions of the electrophoretic length of each capillary, for example at a position about 30 cm from the respective sample injection end of the capillary, are arranged so that they lie side by side on the same plane with the coating film removed. Then, the capillary arrangement plane is irradiated with a laser beam from one side to irradiate the plurality of capillaries with the laser beam simultaneously. In the following description, the capillary arrangement plane can also be referred to simply as the arrangement plane. When a laser beam is passed through, the fluorescence-labeled DNA, which electrophoretically migrates in each of the above capillaries, is excited by laser irradiation and emits fluorescence. Here the DNA is marked with fluorescent substances in four colors, depending on the terminal type of base A, C, G or T. Thereby, the laser irradiation positions of the respective capillaries become light emitting points, and a plurality of light emitting points are arranged at intervals of p on a straight line. In the following, this is referred to as a light emitting point arrangement. When M is the number of light-emitting points (number of capillaries), the total width AW corresponds to the light-emitting point array AW = p * (M-1). For example, if p = 0.36 mm and M = 24, AW = 8.28 mm. A fluorescence detection device recognizes the respective light beams, which are emitted from the light-emitting dot array, together while it spectroscopically separates the light beams. A configuration of the system is in JP-A-2007-171214 (PTL 1).

First, the respective emitted light beams are converted into parallel light beams by a common condenser lens. In the following, the term “jointly” is used in the sense of an M-to-1 correspondence with the meaning that an optical element is provided for a plurality of light-emitting points (M light-emitting points). In contrast, the term “individually” is used in the sense of a 1-to-1 correspondence with the meaning that an optical element is used for a light-emitting point. Here AW <f and AW <D1, where f1 is the focal length and D1 is an effective diameter of the common condenser lens. The condenser lens is, for example, a camera lens with fl = 50 mm and D1 = 36 mm. Then the parallel light beams pass through a long-pass filter to filter a wavelength of the laser beam, and then pass through a common transmission diffraction grating in the direction of the longitudinal axis of each capillary, that is, in the direction orthogonal to both the arrangement direction of the light-emitting point arrangement and the optical Axis of the common condenser lens to be subjected to wavelength dispersion. If DG is the effective diameter of the common transmission diffraction grating, then D1 DG in order not to lag behind in detection performance. For example DG = 50 mm. Then the image of the respective parallel light beams is imaged on the two-dimensional sensor through the common imaging lens. If f2 is the focal length of the common imaging lens and D2 is the effective diameter, D1 D2 must be here so that the detection performance does not decrease. The imaging lens is, for example, a camera lens with f2 = 50 mm, D2 = 36 mm. According to the above, it is possible to collectively acquire wavelength dispersion spectra of the respective light beams emitted from the light emitting dot array. Finally, a change over time of respective wavelength dispersion spectra is analyzed in order to obtain a change over time in the intensity of fluorescence in four colors and to determine the sequence of types of bases, that is, the base sequence.

Another means of detecting fluorescence in four colors at the same time is shown in NPL 1. First, a light beam emitted from a light-emitting surface is converted into a parallel light beam by a condenser lens (here an objective). Here, AW <f and AW <D, where AW is the total width of the light-emitting surface, f is the focal length of the lens and D is the effective diameter. The The lens used is a UPLSAP0 60X W, a product of Olympus, and AW = 0.44 mm, fl = 3 mm and D1 = 20 mm. Then, the parallel light beam is divided into four parallel light beams of four colors by a set of three kinds of dichroic mirrors. Then, through a set of four imaging lenses, images of respective parallel light beams are generated on four two-dimensional sensors. Here, D1 D2 must be so that the detection performance does not decrease, where D2 is the effective diameter of each imaging lens. According to the above, it is possible to collectively capture four partial images of the light-emitting surface in four colors.

Another means for simultaneously detecting light beams emitted from a light emitting dot array is shown in FIG JP-A-2011-59095 (PTL 2). First, respective light beams emitted from a light emitting dot array are converted into individual parallel light beams by a condenser lens array. Here, when p is the distance between the light emitting points and M is the number of the light emitting points, AW = p * (M-1) is the total width of the light emitting point array. If D is the effective diameter of each condenser lens, then D <AW. This configuration differs from that in PTL 1 and NPL 1. With Dl <p, a condenser lens arrangement can be provided in which the respective condenser lenses lie on a straight line. Then the respective parallel light beams are incident on respective individual sensors of the sensor arrangement. According to the above, it is possible to collectively detect the intensities of light beams emitted from the light emitting dot array.
The pamphlet JP 2011-59095 A discloses a light detection device with a structure similar to the present invention and reduced space and energy consumption. In the pamphlet JP 2001-124736 A a capillary electrophoresis device with a similar structure is also described. Devices for multiplex capillary electrophoresis with multiple separation channels are in DE 10 2006 058 575 A1 , WO 2004/017 061 A1 , U.S. 5,324,401 A , U.S. 5,790,727 A , U.S. 5 582 705 A , US 6 531 044 B1 , US 6 191 425 B1 , US 2003/0 116 436 A1 and JP 2001-264 293 A described, wherein in the device of WO 2004/017 061 A1 the light again only traverses the canal walls and the canal walls and the interior of the canal on separate paths. Further light detection-based analysis devices for biological samples are in US 7 029 628 B2 , EP 1 932 594 A1 , U.S. 3,916,197 A and especially too U.S. 5,312,535 A for capillary electrophoresis disclosed.

List of reference literature

Patent literature

• PTL 1: JP-A-2007-171214
• PTL 2: JP-A-2011-59095

Non-patent literature

NPTL 1: Rev Sci Instrum, 2011 Feb; 82 (2): 023701.

Summary of the invention

Technical problem

The light emitting detection device of PTL 1 has a high light condensing performance (light condensing performance of the common condenser lens) and high detection performance (total efficiency of emitted light which contributes to fluorescence detection by the sensor based on the light condensing performance, the permeability of the long-pass filter, the diffraction performance of the diffraction grating, etc. ) of the light rays emitted by the light-emitting points and also a high spectroscopic accuracy of the diffraction grating. The focusing power can be expressed by the f-number F of the common condenser lens and is proportional to 1 / F ² . For example, in the case of a camera lens with f1 = 50 mm and D1 = 36 mm, the f-number is F = f1 / D1 = 1.4, and the light emission detection can be performed with high sensitivity. Since the light beams emitted from the light-emitting points are also imaged with a common imaging lens and recognized by a two-dimensional sensor, the emitted light beams do not mix with one another. That is, the emitted light beams can be recognized independently of one another with little crosstalk. The feature of the above high sensitivity and low crosstalk is particularly important in this technical field in order to accurately detect trace amounts of an object to be measured or to detect a plurality of different objects to be measured simultaneously and independently of each other. The present However, the light emitting detection device has two common lenses and has a relationship in which AW <f and AW <D1 <D2. Therefore, when AW is constant, there is a problem that the overall size of the system is very large and the manufacturing cost of the system is high. For example, in a case where the camera lens is used with f1 = f2 = 50 mm, D1 = D2 = 36 mm, the total size of the light emitting detection device becomes larger than the volume of one column ( 1 , 6 * 10 ⁶ mm ³ ) with a diameter of 100 mm and the height of 200 mm. In the present specification, a total size of the light emitting detection device corresponds to a volume occupied by the optical system from a light emitting point to a sensor, and a volume occupied by the sensor itself is not included in the total size. AW << f1 and AW << D1 are not possible (because this setting would require an enormous camera lens), and there is a problem that the detection performance of the light-emitting point that is away from the optical axis (the light-emitting point that is near one end of the light-emitting dot array), decreases in comparison with the detection performance of the light-emitting point near the optical axis (the light-emitting point located in the center of the light-emitting dot array), and fluctuation in detection sensitivity occurs among the light-emitting dots.

However, no means for solving these problems has been realized heretofore, that is, means for miniaturizing and reducing the cost of a system that detects and simultaneously discriminates light beams of four colors emitted from a light emitting dot array and for reducing fluctuation in detection sensitivity of each emitted light rays have not yet been realized. The miniaturization of the light-emitting detection device would make it possible to install or transport DNA sequencers with capillary arrays in a small area. Alternatively, the user-friendliness of the device can be increased. The number of components in the detection device would also be reduced, or the size of each component would be reduced, thereby lowering the manufacturing cost of the device. Further, the fluctuation in the detection sensitivity of the respective light emitting points would be reduced, thereby enabling quantitative comparison of samples analyzed in the respective capillaries and improving a dynamic range and an overall detection sensitivity of the light emitting point array. This means that the DNA sequencer with capillary array can be used more widely and used.

The use of the light-emitting detection device described in NPL 1 can similarly enable the simultaneous fluorescence detection of the light beams in four colors emitted by a light-emitting point array. However, since the objective lens used in NPL 1 has a total width AW = 0.44 mm, for example only part of the total width (= 8.28 mm) of the light-emitting dot arrangement can be recognized. As with the capillary array DNA sequencer, one common condenser lens and four common imaging lenses are used instead of one objective lens and four. In this case, when DM is the effective diameter of the three kinds of dichroic mirrors, the dichroic mirrors are arranged to be inclined by 45 degrees with respect to the parallel rays of light. In order that the detection performance does not lag behind, √2 * D1 ≤ DM is required. For example DM = 71 mm. Even if there are no four two-dimensional sensors, the overall size of the light emitting detection device is larger than that of PTL 1, and the manufacturing cost increases accordingly. In addition, the space occupied by four two-dimensional sensors is large and the cost involved is very high. The problem of sensitivity fluctuation among light emitting points also remains unsolved.

On the other hand, when using the light emitting detection device disclosed in PTL 2, it is possible to downsize the device because D1 <AW. In a case where light emitted from the light emitting dot array is converged for detection by a condenser lens array as in this literature, it is difficult to obtain both high sensitivity performance and low crosstalk performance because both are in conflict with each other. In PTL 2 there was no discussion or reflection on these problems and how to solve them. These problems are described in detail below.

As described above, the light converging power can be expressed by F = f / D (the light converging power is proportional to 1 / F ² ), where f is the focal length of each condenser lens and D is an effective diameter. If it is assumed here that D is constant, the light condensing power appears to increase as the focal length f becomes smaller, that is, the closer the light emitting point and the condenser lens are to each other. This is true when the light emitting point is an infinitely small size, but not necessarily when the light emitting point is a finitely small size. If the diameter of the light emitting point is defined as d, the infinitely small size means a case where d << f or d ≈ 0, and the finitely small size means a case where d> 0. In practice, the finitely small size corresponds to a case where d ≥ 0.01 mm. In the following, a case in which the size of the light emitting point is finite is considered in the present invention. When the center of the light emitting point is in the focal position of the lens, the light emitted from the center of the light emitting point is converted into a parallel light beam by the lens, and the light beam propagates in a direction parallel to the optical axis along the optical axis of the lens out. On the other hand, light emitted from the end of the light emitting point is converted into a parallel light beam by the condenser lens, and the light beam propagates in a direction having an angle θ = tan ^-1 (d / 2 / f) with the optical axis the lens forms. That is, these rays of light move away from the lens. So, when light emission is detected by placing a sensor the size of the parallel light beam emitted from the center of the light emitting point at a position on the optical axis of the lens that is a certain distance from the lens (or assigned to part of an area sensor), all of the light emitted from the center of the light emitting point can be detected. In contrast, only part of the light emitted from the end of the light emitting point can be recognized. Since the angle θ decreases with increasing focal length f, the proportion of detectable light emission increases.

From the above it can be seen that under certain aspects it is better to set f small, while under other aspects it is better to set f large in order to increase the light bundling power and to obtain a high sensitivity, which is why there is a trade-off between these aspects . However, studies on the optimal value of f for obtaining high sensitivity have not yet been made, even in PTL 2. As described below, in the present invention, as a result of carrying out this study under practical conditions, it was found that the total light condensing performance is related the smaller the f increases. This suggests that the effect of decreasing the focal length f on increasing the amount of detection light of the light emitted from the center of the light emitting point is greater than the effect of increasing the focal length f on increasing the amount of detection light of the light emitted from the end of the light emitting point .

For crosstalk, however, the discussion must be conducted separately from the above light bundling power. Since the angle θ increases as f decreases, the proportion of light emitted from the end of the light emitting point that is from an adjacent sensor (or an area assigned to part of an area sensor) located around that from the adjacent light emitting point increases To detect light emitted from the point, therefore, the crosstalk increases when the light emitted from the adjacent light-emitting point is detected. That is, it has been found that there is a conflict between the aspect of decreasing f to increase the light converging performance and the aspect of increasing f to obtain high sensitivity. So far, however, no study has been carried out on which f is optimal to reconcile high sensitivity and low crosstalk, not even in PTL 2.

the solution of the problem

The light emitting detection device according to the present invention is defined in the independent claims. Further advantageous designs are described in the dependent claims.

Advantageous Effects of the Invention

According to the present invention, it is possible to miniaturize the device which detects light emitted from the light emitting dot array with high sensitivity and little crosstalk, and to reduce the overall size of various devices using the system. This makes it possible to reduce the space required by the devices, to transport the system or devices, and to improve the operability of the devices. The number of components composing the device is reduced and the component itself is miniaturized, making it possible to reduce the manufacturing cost.

Other problems, configurations and effects will be apparent from the description of the following embodiments.

Figure list

• [ 1 ] 1 Fig. 13 is a schematic representation of a configuration in which light emitted from a light emitting point is converged by a condenser lens to give a light beam.
• [ 2 ] 2 Fig. 13 is a diagram showing a relationship between an optical path length g from a condenser lens to a sensor and a relative amount of detection light using the focal length f of the condenser lens as a parameter.
• [ 3 ] 3 Fig. 13 is a schematic illustration of a configuration in which light emitted from two adjacent light emitting points is each converged by the condenser lens to give separate light beams.
• [ 4th ] 4th Fig. 13 is a schematic illustration of a configuration in which light emitted from two adjacent light emitting points is each converged by the condenser lens to give a mixed light beam.
• [ 5 ] 5 Fig. 13 is a diagram showing a relationship between an optical path length g from the condenser lens to the sensor and the focal length f of the condenser lens which satisfies the conditions of high sensitivity and low crosstalk.
• [ 6th ] 6th Fig. 13 is a schematic diagram of a light-emitting detection device in which light emitted from a light-emitting dot array is converged by a condenser lens assembly, the converged light being incident in parallel on a color sensor to be detected.
• [ 7th ] 7th Fig. 13 is a schematic representation of a light-emitting detection device in which light emitted from a light-emitting dot array is each bundled by a condenser lens array, the light beams are split into different wavelength bands in parallel with a dichroic array and are incident on a sensor in parallel.
• [ 8th ] 8th Fig. 13 is a schematic representation of a light-emitting detection device in which light emitted from a light-emitting dot array is converged by a condenser lens array, parallel wavelength dispersion is performed on the light rays with a wavelength dispersion element, and a respective image is generated with an imaging lens array.
• [ 9 ] 9 Fig. 13 is a schematic representation of a light-emitting detection device in which light emitted from a light-emitting dot array is each converged by a condenser lens array, the light beams are deflected in parallel with an optical element and are incident on a sensor in parallel.
• [ 10 ] 10 Fig. 13 is a schematic representation of a light-emitting detection device in which light emitted from a light-emitting point array is each bundled by a condenser lens array, the light beams are deflected in parallel with an optical fiber array, and the light beams emitted by the optical fiber array are incident in parallel on a sensor.
• [ 11 ] 11 Fig. 13 is a schematic diagram showing a configuration example of a capillary array DNA sequencer.
• [ 12th ] 12th Fig. 13 is a diagram showing an example of dichroic arrangement in which light rays incident in parallel on the dichroic arrangement are split in a vertical direction, and showing the result of calculating parallel light rays of maximum divisible width.
• [ 13th ] 13th Fig. 13 is a diagram showing an example of a dichroic arrangement in which light rays incident vertically on the dichroic arrangement incident, divided in the same direction, and which shows the result of calculating parallel rays of light with maximum divisible width.
• [ 14th ] 14th Fig. 13 is a diagram showing an example of a compacted dichroic array in which light rays vertically incident on the dichroic array are split in the same direction, and showing the result of calculating parallel light rays with the maximum divisible width.
• [ 15th ] 15th Fig. 13 is a diagram showing an example of compacted and staggered dichroic array in which light rays vertically incident on the dichroic array are split in the same direction, and showing the result of calculating parallel light rays of maximum divisible width.
• [ 16 ] 16 Fig. 13 is a diagram showing a configuration of a best mode of embodiment of the dichroic array that shortens an optical path length and increases an opening width.
• [ 17th ] 17th Fig. 13 is a diagram showing the relationship between a thickness β of a dichroic mirror and a dichroic array pitch x, where an optical path length _{corresponds to a dichroic array L max} or less and an opening width W _min or more.
• [ 18th ] 18th Fig. 13 is a diagram showing a relationship among a dichroic array distance x, an opening width W and a change in optical path length ΔL.
• [ 19th ] 19th FIG. 13 is a diagram showing a relationship between the step-like displacements y and z of a dichroic arrangement and an opening width W. FIG.
• [ 20th ] 20th Fig. 13 is a diagram showing an example of a dichroic arrangement in which light rays vertically incident on the dichroic arrangement are split in the same direction with a dichroic mirror inclined more than 45 degrees from the vertical direction, and a Represents the result of the calculation of a parallel light beam with a maximum divisible width.
• [ 21 ] 21 Fig. 13 is a diagram showing a relationship between an inclination θ _{0 of} a dichroic mirror and an opening width W of a dichroic array.
• [ 22nd ] 22nd 13 is a schematic representation of a light-emitting detection device in which a portion of the light emitted by a light-emitting dot arrangement is transmitted through a pinhole arrangement, the transmitted light is in each case bundled by a condenser lens arrangement and the light is incident in parallel on a sensor.
• [ 23 ] 23 Fig. 13 is a schematic representation of a light-emitting detection device, in which a portion of the light emitted from a light-emitting dot array which is arranged at smaller intervals than the condenser lens array is transmitted through a pinhole arrangement, the transmitted light is bundled in each case by a condenser lens arrangement, and is incident on a sensor in parallel.
• [ 24 ] 24 Fig. 13 is a schematic representation of a light-emitting detection device, in which a portion of the light emitted from a light-emitting dot array is transmitted through a pinhole array, the transmitted light is each converged by a condenser lens array to form light rays, the light rays having a dichroic Arrangement can be divided into different wavelength bands in parallel and incident on a sensor in parallel.
• [ 25th ] 25th Figure 3 is a schematic illustration of the imaging of a light emitting area by a light emitting detection device comprising a pinhole assembly, a condenser lens assembly, and a scanning mechanism.
• [ 26th ] 26th Fig. 13 is a schematic sectional view showing a configuration example of an apparatus in which a V-groove arrangement having a plurality of capillaries and a single condenser lens arrangement are integrated.
• [ 27 ] 27 Fig. 13 is a schematic sectional view showing a configuration example of an apparatus in which a V-groove assembly having a plurality of capillaries, a pinhole assembly, and a single condenser lens assembly are integrated.
• [ 28 ] 28 Fig. 13 is a schematic sectional view showing a configuration example of an opaque device in which a V-groove assembly having a plurality of capillaries and a pinhole assembly are integrated, in connection with a device in which a single condenser lens assembly is integrated.

Description of embodiments

1 Fig. 13 is a schematic sectional view including an optical axis of an optical system extending from a light emitting point 1 emitted light using a condenser lens 2 bundles and detects it by a sensor arranged at a certain distance from the condenser lens. In the present invention, the terms “light emitting point” and “emitted light” are used frequently, but this does not necessarily mean that the fluorescence, phosphorescence, etc. is emitted from the substance to be detected itself, but is often scattered light generated by the detection object will when it is irradiated with light, or transmitted light transmitted by the detection object, which is why it is collectively referred to as light emitted from the light emitting point.

"D" is used as the diameter of the light emitting point 1 defines "f" as the focal length of the condenser lens 2 , "D" as the effective diameter of the condenser lens 2 , “D” as the diameter of a detection area of the sensor, and “g” as the optical distance (optical path length) between the condenser lens 2 and the sensor. When the distance between the light emitting point 1 and the condenser lens 2 is set to f, that is, when the center of the light emitting point 1 in the focal position of the condenser lens 2 forms the light emanating from the center of the light emitting point 1 is emitted through the condenser lens 2 a parallel beam of light 3 , which has a diameter D and spreads in the direction of the optical axis. In the sensor, which is arranged at a position which is the optical path length g from the condenser lens 2 is away, generates the parallel beam of light 3 a point of light 4th with a diameter D. 1 shows the light emitting point 1 on an underside of the optical system, seen in the direction of the optical axis, and the light point 4th as well as a point of light 5 (described below) on its upper side, seen in the direction of the optical axis. On the other hand, the light emitted from a left end of the light emitting point 1 is emitted through the condenser lens 2 into a parallel beam of light 3 ' which has a diameter D and propagates in the direction of an angle θ to the optical axis. The parallel light beam is generated in the sensor 3 ' the point of light 5 with a diameter D.

The condensing power of light emitted from the center of the light emitting point 1 is proportional to 1 / F ² , where F is a f-number F of the condenser lens 2 designated. If D is constant, the smaller the focal length f, the higher the light condensing power, since F = f / D. On the other hand, the point of light shifts 5 that of the left end of the light emitting point 1 emitted light from the detection area of the sensor to the right. That is, the point of light 4th is fully recognized, the point of light 5 however, it is recognized to the extent that the point of light 5 the point of light 4th overlaps. As the overlap area increases, the amount of light that can be seen over the entire area of the light emitting point increases. For this reason, the angle θ of the parallel light beam is 3 ' preferably small to the optical axis. Since θ = tan ^-1 (d / 2 / f), it is better if the focal length f is small when d is constant. As described above, it is to increase the amount of detection light from the light emitting point 1 under certain aspects better if f is small, while under other aspects it is better if f is large. Although there is a trade-off between these aspects, no studies on the optimal range of f have been performed. The conditions of f and g for increasing the amount of detection light from the light emitting point 1 are therefore explained below.

The in 3 the fluorescence detection device shown in PTL 1 is taken as a reference. In a typical fluorescence detection device, the focal length f1 of a common condenser lens is f1 = 50 mm, and the effective diameter D1 is D1 25 mm. The f-number F of the lens is F = f1 / D1 2.0. If, when using a condenser lens with F ₀ = 2.0, light emitted from a light-emitting point of infinitely small size and focused at a focal point of the lens is converted by the lens into a parallel light beam and the parallel light beam is detected by the sensor without loss , an amount of detection light is taken as a reference value (100%). In the following, the amount of detection light for each light-emitting point of infinitely small size is evaluated by a relative amount of detection light relative to the reference value. It is assumed that the light-emitting point, which has a finitely small size with a mean effective diameter d, is composed of a plurality of light-emitting points each having an infinitely small size. In this patent specification, a “light-emitting point of finitely small size” is simply referred to as a “light-emitting point”, and a “light-emitting point of infinitely small size” is always referred to as “light-emitting point of infinitely small size”. The relative detection light amount of the light emitting point is set as an average of the relative detection light amounts of a large number of light emitting points of infinitely small size that make up the light emitting point. In the above example, for example, the light condensing power is increased (F ₀ / F) ² = 2.0 times when the condenser lens is replaced with one with F = 1.4, and therefore the detection light amount of the light emitting point of infinitely small size is 200% . Here, it is assumed that the total amount of light emitted in all directions from the light-emitting point is constant, and that the luminance inside the light-emitting point is spatially uniform. In the present example of a typical fluorescence detection device, the distance between the light-emitting points of the light-emitting point arrangement is p = 0.36 mm, the number of light-emitting points M = 24, and the total width of the light-emitting point arrangement AW = p * (M-1) - 8.28 mm. Since the light-emitting point is located in the center of the light-emitting point array near the focal point of the lens, the relative amount of detection light from the light-emitting point is almost 100%. And there the light emitting point at both ends of the light emitting point array is away from the focal point of the lens, the relative amount of detection light from the light emitting point decreases to about 50%. The present invention aims to increase the relative detection light amount of each light emitting point to more than 50% so that the detection sensitivity of each light emitting point is at least as high as that of the prior art.

2 FIG. 13 is a diagram illustrating the calculation results of a relationship between g and a relative detection light amount, wherein in FIG 1 configuration f shown is used as a parameter. Here is the mean effective diameter of the light emitting point 1 set to d = 0.05 mm. The effective diameter of each condenser lens 2 was set to D = 0.5 mm. The relative amount of detection light was calculated taking into account the brightness of the lens F = f / 0.05. The light emitting point 1 with the effective diameter d = 0.05 mm is composed of about 500 light-emitting points of infinitely small size arranged at intervals of 0.1 µm, and the relative amount of detection light for each light-emitting point of infinitely small size becomes as in FIG 1 based on the overlap area ratio of the light spot 4th and the point of light 5 calculated. The relative amount of detected light from the light emitting point 1 is obtained by averaging the relative amounts of detection light of all light-emitting points of infinitely small size. As a result, as in 2 first found out that the relative detection light quantity is greater, the smaller f or the smaller g. This suggests that the effect of decreasing f is to increase the relative amount of detection light from a light-emitting point of infinitely small size, that at the center of the light-emitting point 1 is greater than the effect of increasing f to increase the overlap area ratio. This also suggests that the effect of decreasing g to increase the overlap area ratio at each focal point f is large.

In 1 and 2 is the center of the light emitting point 1 at the focal point of the condenser lens 2 arranged, and the light is emitted in a parallel beam of light. The present invention also functions satisfactorily under these conditions, but as will be described in detail below, it has been found that a better condition for reconciling high sensitivity with low crosstalk is when the center of the light-emitting point 1 slightly from the focal point of the condenser lens 2 is offset and the emitted light is converted into a slightly restricted light beam 6. That is, the optimum condition has been found to be when the image of the light emitting point 1 at the sensor position, that is, at a position of the optical path length g from the condenser lens 2 is mapped as the diameter of the light emitting point 1 can be minimized.

3 Fig. 3 is a sectional view with the optical axis of the optical system that includes each condenser lens 2 each bundles light from two adjacent light-emitting points 1 is emitted, and in each case images at the sensor position 7th receives the images of the light emitting points 1 are. In the present invention, the term "image of the light emitting point" does not necessarily mean an image on which the light emitted from the light emitting point is focused, but generally a cross section at a certain position of a light beam of the light emitted and converged from the light emitting point. The diameter of the light emitting point 1 is defined as "d", the focal length of the condenser lens 2 as "f", the effective diameter of the condenser lens 2 is as "D", the distance between the light emitting points 1 and a distance between the condenser lenses 2 as “p”, the diameter of the detection area of the sensor as “D”, the optical distance (optical path length) between the condenser lenses 2 and the sensor as "g", and the diameter of the image 7th of the light emitting point at the sensor position as "d". When the distance between the light emitting point 1 and the condenser lens 2 is set and that from the light emitting point 1 emitted light through the condenser lens 2 focused on the sensor position is the diameter d 'of the image 7th of the light emitting point is minimized.

ist der Durchmesser d' des Bilds 7 des lichtemittierenden Punkts dabei wie folgt: $$\text{D}\text{'}\ge \text{m}*\text{d}=\left(\text{g}-\text{f}\right)/\text{f}*\text{d}$$

Since the image magnification is expressed as follows: $$\text{M.}=\left(\text{G}-\text{f}\right)/\text{f},$$

is the diameter d 'of the image 7th of the light-emitting point as follows: $$\text{D.}\text{'}\ge \text{m}*\text{d}=\left(\text{G}-\text{f}\right)/\text{f}*\text{d}$$

Here, equation (2) expresses an equal sign when that from the light emitting point 1 emitted light through the condenser lens 2 is focused at the sensor position, otherwise it expresses an inequality sign. 3 represents the light emitting point 1 on a bottom of the optical System, seen in the direction of the optical axis, and the image 7th of the light emitting point on its upper side, viewed in the direction of the optical axis. The present invention works satisfactorily when g 2 * f, that is, when m 1. Preferably, a good condition is obtained when m 5 or m 10. Therefore, as shown in FIG 2 seen because it is necessary to reduce the focal length “f” to the mm plane in order to increase the relative amount of detection light, but it is physically difficult to reduce the optical path length “g” so much.

In each of the drawings of this specification are the light emitting point 1 and the picture 7th of the light-emitting point are each shown as circular, but they can actually have other shapes without being limited to the circular shape. Generally, d is the diameter of the light emitting point 1 and the diameter d 'of the image 7th of the light emitting point are widths in a direction of arranging the light emitting point, respectively 1 and the picture 7th of the light emitting point. Furthermore, as described further below, a plurality of optical distances (optical path lengths) between the condenser lens can be used in the same light-emitting detection device 2 and the sensor. In this case, the above equations (1) and (2) and the following equations (3) to (18) can be established assuming that the optical distance between the condenser lens 2 and the sensor is generally set to an optical path length "s" and the maximum value of s is set to the optical path length g. The reason for this is described below. When the optical distance between the condenser lens 2 and is represented to the sensor by a variable s, then 0 ≤ s ≤ g, where g is the maximum value. Where is the diameter d '(s) of the image 7th of the light emitting point for each variable s is smaller than the larger of d '(s = 0) = D and d' (s = g) = d '. That is, if D ≥ d ', d' (s) ≤ D; and if D ≤ d ', d' (s) ≤ d '. In the first case, no crosstalk occurs, and if in the second case the criterion of crosstalk is met with s = g, this means that the same criterion itself can be met if 0 s g. On the other hand, the relative amount of detected light is constant regardless of the optical path length "s".

In the description, the condenser lens has 2 essentially a circular shape with an effective diameter D as the diameter, but this is not necessary. In general, there is the effective diameter D of the condenser lens 2 the arrangement direction of the light emitting point 1 and a width of the condenser lens 2 in the arrangement direction, and a width in a direction orthogonal to the arrangement direction is not limited to this. The condenser lens 2 can be circular, elliptical, square, rectangular, or otherwise shaped. In the above explanation, the diameter d '(s = 0) = D of the image 7th of the light emitting point as a diameter in the arranging direction of the light emitting point 1 and the arrangement direction of the condenser lens 2 to be viewed as. The diameter of the image 7th of the light emitting point in the direction orthogonal to the arrangement direction does not contribute to crosstalk regardless of how large it is. If g is large enough, d '(s = g) = d' is irrelevant to D. Therefore, the crosstalk conditions of the following equations (13) to (18) become independent of the width of the condenser lens 2 set in the direction orthogonal to the arrangement direction. When the width of the condenser lens 2 in the direction orthogonal to the arrangement direction is larger than the effective diameter D, on the other hand, the f-number F can be smaller than F = f / D, that is, the light condensing power can be increased. In this case, the conditions of the following equations (3) to (12) in terms of sensitivity can give an even higher relative detection light amount and sensitivity.

First, the conditions for obtaining high sensitivity will be considered. The light condensing power of the light emitting point 1 emitted light through the condenser lens 2 can be determined by the f-number F of the condenser lens 2 and F = f / D can be expressed. In order to achieve the relative detection light amount of 50% or more, it may be necessary to set F 2.8, that is, f 2.8 * D. On the other hand, the following equation satisfies the condition that the relative amount of detection light is 50% or more because it is necessary to set p D in order to configure the condenser lens assembly. $$\text{F.}\le 2.8*\text{p}$$

$$\text{F}\le \mathrm{1,4}*\text{p}$$

$$\text{F}\le \mathrm{1,0}*\text{p}$$

$$\text{F}\le \mathrm{0,7}*\text{p}$$

Accordingly, the relative detection light amount becomes 100 under the conditions of F 2.0, 1.4, 1.0 and 0.7, that is, the following equations (4), (5), (6) and (7) % or more, 200% or more, 400% or more, and 800% or more. $$\text{F.}\le 2.0*\text{p}$$

$$\text{F.}\le 1.4*\text{p}$$

$$\text{F.}\le 1.0*\text{p}$$

$$\text{F.}\le 0.7*\text{p}$$

$$\text{F}\le \left(1/\left(\mathrm{2,0}*\text{p}\right)+1/\text{g}\right)-1$$

$$\text{F}\le \left(1/\left(\mathrm{1,4}*\text{p}\right)+1/\text{g}\right)-1$$

$$\text{F}\le \left(1/\left(\mathrm{1,0}*\text{p}\right)+1/\text{g}\right)-1$$

$$\text{F}\le \left(1/\left(\mathrm{0,7}*\text{p}\right)+1/\text{g}\right)-1$$

The above equations (3) to (7) are satisfied when the distance between the light emitting point 1 and the condenser lens 2 can be approximated by f, but can be expressed more precisely as follows. Since f2 / (gf) + f is the distance between the light emitting point 1 and the condenser lens 2 is if that from the light emitting point 1 emitted light through the condenser lens 2 is focused on the optical path length g, the effective f-number F of the condenser lens 2 can be expressed as F '= (f2 / (gf) + f) / D. Accordingly, under the precise conditions of the following equations (8), (9), (10), (11), and (12), the relative detection light amount becomes 50%, 100% or more, 200% or more, 400% or more and set 800% or more. $$\text{F.}\le \left(1/\left(2.8*\text{p}\right)+1/\text{G}\right)-1$$

$$\text{F.}\le \left(1/\left(2.0*\text{p}\right)+1/\text{G}\right)-1$$

$$\text{F.}\le \left(1/\left(1.4*\text{p}\right)+1/\text{G}\right)-1$$

$$\text{F.}\le \left(1/\left(1.0*\text{p}\right)+1/\text{G}\right)-1$$

$$\text{F.}\le \left(1/\left(0.7*\text{p}\right)+1/\text{G}\right)-1$$

$$\text{V}\le 12*\text{p/d'}$$

Conditions for obtaining low crosstalk will next be considered. If the pictures 7th of the neighboring light emitting points 1 do not overlap each other, as in 3 shown, there is no crosstalk. If the pictures 7th adjacent light emitting points 1 however, as in 4th overlap as shown, crosstalk occurs. In the following, the crosstalk is represented by a ratio X of the overlapping area of the image 7th adjacent light emitting points to the surface of the image 7th of the light emitting point in 4th expressed. The crosstalk is set to X or less under the following conditions: $$\text{X}=1/*\left({\text{cos}}^{-1}\left({\text{<figure-callout id="2" label="V" filenames="DE112016007594B3\_0062.png,DE112016007594B3\_0065.png" state="{{state}}">V</figure-callout>}}^2/2-1\right)-\text{sin}\left({\text{cos}}^{-1}\left({\text{<figure-callout id="2" label="V" filenames="DE112016007594B3\_0062.png,DE112016007594B3\_0065.png" state="{{state}}">V</figure-callout>}}^2/2-1\right)\right)\right)$$

$$\text{V}\le \mathrm{12th}*\text{p / d '}$$

When equation (14) is converted from equation (2), it can be expressed as follows: $$\text{F.}\ge \text{1/}\left(\left(2*\text{p}\right)/\left(\text{V}*\text{d}\right)+1\right)*\text{G}$$

To perform the detection of the light, that from the light emitting point to be detected 1 is emitted without being influenced by light that from the light-emitting points adjacent on both sides 1 must be the distance between the two images 7th of the light emitting point in 4th be at least greater than the radius (or half the diameter) of the image of the light emitting point. When equations (13) and (14) give X = 0.39 (39%) and V = 1, equation (15) can be expressed as follows: $$\text{F.}\ge \text{1/}\left(2*\text{p}/\text{d}+1\right)*\text{G}$$

To that of a multitude of light emitting points 1 To detect emitted light more effectively and independently, it is desirable to set the total crosstalk ratio from both sides to 50% or less. For this purpose, if equations (13) and (14) give X = 0.25 (25%) and V = 1.27, equation (15) can be expressed as follows: $$\text{F.}\ge 1/\left(\left(2*\text{p}\right)/\left(1.27*\text{d}\right)+1\right)*\text{G}$$

The crosstalk is preferably set to 0%. For this purpose, when equations (13) and (14) give X = 0 (0%) and V = 2, equation (15) can be expressed as follows: $$\text{F.}\ge 1/\left(\text{p}/\text{d}+1\right)*\text{G}$$

As described above, it is possible to obtain a desired relative detection light amount and sensitivity by selecting g and f such that one of equations (3) to (12) is satisfied for given p and d. On the other hand, it is possible to obtain a desired crosstalk by choosing g and f such that one of the equations (16) to (18) is satisfied for the given p and d. That is, it is possible to maintain both the relative detection light amount and the crosstalk, which are in conflict with each other, at the desired level by selecting g and f such that one of equations (3) to (12) and one of equations (16) to (18) are satisfied.

5 represents the equations (3) to (12) and (16) to (18) with a horizontal axis g and a vertical axis f for the case p = 1 mm and d = 0.05 mm as a typical example Numbers shown in curves or straight lines indicate boundary lines of the corresponding equations, a symbol ↓ (arrow downward) indicates an area below the boundary line, and a symbol ↑ (arrow upward) indicates an area above the boundary line. For example, in order to make the equation (3) satisfy the condition that the relative detection light amount is 50% or more, g and f can be set in the range below the straight line ↓ (3) in 5 lie. On the other hand, in order to ensure that equation (17) fulfills the condition that the crosstalk is 25% or less, g and f can be used in the area above the straight line ↑ (17) in 5 lie. That is, in order to set the relative detection light amount to 50% or more and the crosstalk to 25% or less, g and f are allowed in the area below the straight line ↓ (3) in 5 and in the area above the straight line ↑ (17) in 5 lie.

As can be seen from the orders of magnitude of g and f, the light-emitting detection device with the in 5 g and f shown, characterized in that through a high sensitivity and a low crosstalk not only increases the power, but also the size of the device compared to the detection device in PTL 1 and NPTL 1 can be reduced by 1 to 3 orders of magnitude. As can be seen from the above, the areas of g and f which satisfy the conditions of high sensitivity and low crosstalk become narrower the smaller p is and the larger d is, and downsizing of the detection light-emitting device is inevitable. Conversely, the smaller p is and the larger d is, the more useful the features of the present invention become, and its advantage over the conventional method becomes significant. In particular, in the case of p 20 mm and preferably p 10 10 mm, the features of the present invention become particularly useful. Further, the features of the present invention become particularly useful in the case of d 0.005 mm, and preferably d 0.01 mm.

Next, a description will be given of a method of performing multicolor recognition based on the above conditions. A color sensor is at the position of the picture 7th of the light emitting point in 3 or 4th arranged such that the sensor surface is perpendicular to the optical axis of the condenser lens 2 stands, that is, the sensor surface is parallel to the plane of arrangement of the light-emitting point 1 and to the plane of arrangement of the condenser lenses 2 . Here, the color sensor is configured to provide two or more kinds of pixels capable of distinguishing and recognizing at least two kinds of light having different wavelengths. The most common color sensor is a color sensor used in consumer digital cameras. In the color sensor, three types of pixels are arranged two-dimensionally to distinguish the three colors R, G and B, that is, red, green and blue. Such an ordinary color sensor can also be used in the present invention. In recent years, ordinary color sensors have achieved high sensitivity and can also be used in this technical field. The above color sensor is best for distinguishing three colors, but it can also distinguish four or more colors based on the intensity ratio of the three types of pixels, such as which is the case with digital cameras. Therefore, it is possible to apply the detection device with the above-described color camera to a DNA sequencer that performs four-color recognition.

To be able to distinguish the exact color from the light emitting point 1 however, it is effective to determine the diameter d 'of the image 7th of the light emitting point larger than the size of each type of pixel. This is because it is possible to suppress the influence of a fluctuation in the relative position of the image 7th of the light emitting point and the pixel to be avoided by reducing the light emission for each light emitting point 1 and is detected with a plurality of pixels for each kind of pixel. Under the appropriate conditions of the present invention given in equations (3) through (12) and (16) through (18), m> 1 in equation (1), that is, d '> d in equation (2) ) is often met, and therefore it is easy to meet the above conditions. Because the plane of arrangement of the light emitting point 1 and the plane of arrangement of the condenser lens 2 are parallel to the sensor surface, it is also possible, the sensor surface of the plane of arrangement of the condenser lens 2 and the conditions of equations (3) to (12) and (16) to (18) can be easily met.

Recently, color sensors having four kinds of pixels with IR (infrared) in addition to R, G and B are commercially available, and it is effective to use such a color sensor for four-color recognition in a DNA sequencer. The different types of pixels can be arranged on the same plane or in a direction perpendicular to the sensor surface. As described above, the application of the practically used color sensor to the present invention is effective to keep the development cost down. It goes without saying that it is effective to adjust the number of kinds of pixels of the color sensor and the characteristics of color differentiated from each kind of pixels according to the purpose of use.

6th Fig. 10 illustrates an example of a multicolor detection device using a color sensor. 6 (a) Figure 8 shows the multicolor detection device in a direction perpendicular to the plane defining each optical axis of the condenser lenses 2 contains, and 6 (b) represents one of the two-dimensional color sensor 11 captured image 12. Here, an example is described in which four colors are recognized.

As in 6 (a) is shown by the respective light-emitting points 1 emitted light from a single condenser lens 2 bundled to form a light beam 9, transmitted in parallel by a common long-pass filter 10 and incident on a conventional two-dimensional color sensor 11 parallel one. The long-pass filter 10 is used to block excitation light such as light with a wavelength that hinders multicolor recognition. As in 6 (b) are shown on an image 12 of the two-dimensional color sensor 11 images 7th of the respective light emitting points 1 emitted light generated. The two-dimensional color sensor 11 includes, for example, four kinds of pixels including an A pixel 13 which mainly detects A light emission, a B pixel 14 which mainly detects B light emission, a C pixel 15 which mainly detects C light emission, and a D pixel 16, which detects mainly D light emission, the four kinds of pixels each regularly arranged in a plurality. The size of each pixel 13, 14, 15 and 16 is S = 0.05 mm. When d = 0.05mm, f = 1mm and g = 10mm and the light emitting points 1 are focused on the two-dimensional color sensor, according to equation (1) m = 9 and according to equation (2) d '= 0.45 mm. That is, S <d 'is satisfied, and the image 7th of the light emitting point is recognized with about 64 pixels and about 16 pixels per kind of pixel.

In this way it is possible to have four colors of the light emitting point 1 The light emitted can be identified precisely by looking at the respective images 7th of the light emitting point can be captured by a plurality of pixels of each type. Even if, for example, the relative position between pixels of each type and the image 7th of the light emitting point fluctuates, it does not matter. Alternatively, each of the colors can be recognized uniformly and stably even if the distribution of light intensity in the image 7th of the light emitting point is non-uniform. In multicolor detection device, which in 6th as shown, meet the diameter d of each light emitting point 1 , the distance p between the respective light emitting points 1 and between the respective condenser lenses 2 , the focal length f and the effective diameter D of each condenser lens 2 and the optical path length g between each condenser lens 2 and the sensor 11 any of the equations (3) to (7), the equations (8) to (12) and the equations (16) to (18), the predetermined high sensitivity and the low crosstalk are achieved, and the size and cost of the detection device are reduced.

When using a color sensor in which several types of pixels are arranged on the same plane, as in 6th shown, however, there is a problem that the utilization of the incident light is poor. For example, as in 6th When four colors are distinguished by a color sensor in which four kinds of pixels are arranged, the utilization of incident light becomes 1/4 or less reduced. This can prove problematic for performing light emission detection with increased sensitivity. The utilization of the incident light can be increased by using a color sensor in which a plurality of types of pixels are arranged in the direction perpendicular to the sensor surface, but such a color sensor is not generally used yet. Therefore, a further multicolor detection method with high utilization of the incident light is proposed in the following.

One way to increase the utilization of the incident light is to use one or more dichroic mirrors. A dichroic mirror has a multilayer film formed on a front surface of at least one side of a transparent substrate such as glass to make reflected light and transmitted light having different wavelength bands from light generally incident at 45 °, thereby making both The reflected light as well as the transmitted light can be used to increase the efficiency of the incident light. In general, it is possible with a dichroic mirror to recognize a maximum of two colors, to recognize a maximum of three colors by combining two types of dichroic mirrors, and accordingly to recognize a maximum of (n + 1) colors by combining N types of dichroic mirrors. In addition to the above dichroic mirrors, a band pass filter, a color glass filter or a total reflection mirror is often used in combination. In consumer digital video cameras, three types of dichroic mirrors are combined and three-color recognition is performed with three CCDs. In addition, it is NPTL 1 an example in which three kinds of dichroic mirrors are combined and four-color recognition is performed with four CCDs. As described above, when the dichroic mirror is used, multiple sensors are often used because the reflected light and the transmitted light have different directions. This is in conflict with the reduction in size and cost of the detection device which is an object of the present invention.

7th illustrates an example of a multicolor detection device in which these problems have been solved. 7 (a) Fig. 13 shows the multicolor detection device in a direction perpendicular to a plane defining the respective optical axes of the condenser lenses 2 contains, 7 (b) Fig. 10 shows a cross section of the multicolor detection device with the optical axis of a condenser lens 2 and perpendicular to the direction of arrangement of the condenser lens arrangement, and 7 (c) Figure 29 shows an image taken by a two-dimensional sensor 30th is recognized. Here is an example in which four colors are recognized.

For example, light emitted from respective light-emitting points 1 is emitted from four light emitting dot arrays, each through the single condenser lens 2 bundled and transmitted through the common long-pass filter 10 as a parallel light beam 9. This matches with 6th . Here, a dichroic arrangement is used in which four kinds of common dichroic mirrors 17, 18, 19 and 20 are arranged side by side, as in FIG 7 (b) shown. The light beams 9 are incident in parallel on the respective dichroic mirrors 17 to 20, are split into four light beams 21, 22, 23 and 24 in the direction perpendicular to the direction of the light emitting dot array, and the four light beams propagate in the same direction as the light beam 9, that is, in the direction of the optical axis of the condenser lens 2 , and fall in parallel on a common two-dimensional sensor 30th to give images 25, 26, 27 and 28 of the light emitting points. Here the dichroic mirror 20 can be replaced by a total reflection mirror, but is referred to as dichroic mirror 28 for the sake of simplicity.

As in 7 (c) are shown on the image 29 of the two-dimensional sensor 30th sixteen images 25 through 28 of light emitting points obtained by dividing that of four light emitting points into four 1 emitted light were observed together. Here, among the light beams 9, a light beam transmitted from the dichroic mirror 17 is a light beam 21, a light beam reflected from the dichroic mirrors 17 and 18 is a light beam 26, a light beam reflected from the dichroic mirror 17 , transmitted by the dichroic mirror 18 and reflected by the dichroic mirror 19 is a light beam 23, and a light beam reflected by the dichroic mirror 17, transmitted by the dichroic mirrors 18 and 19 and reflected by the dichroic mirror 20 is a light beam 24. By designing and regulating the transmission and reflection properties of the long-pass filter 10 and the dichroic mirrors 17 to 20, the light beam 21 has mainly a fluorescence component A, the light beam 22 mainly a fluorescence component B, the light beam 23 mainly a fluorescence component D and the light beam 24 mainly a fluorescence component D, and the fluorescence A, B, C and D can be recognized by recognizing the intensity of the images 25, 26, 27 and 28 of the light emitting points.

Although the wavelength bands of the light beams 21, 22, 23 and 24 can be designed arbitrarily, the dichroic mirrors 17 to 20 are easy to design when these light beams are in the order of Wavelengths are arranged. That is, the order may be a central wavelength of A fluorescence> a central wavelength of B fluorescence> a central wavelength of C fluorescence> a central wavelength of D fluorescence, or a central wavelength of A fluorescence <a central wavelength of B fluorescence <a Central wavelength of C fluorescence <be a central wavelength of D fluorescence. Although this is in 7th not shown, a band pass filter or a color glass filter having other spectral characteristics is arranged at at least one position of the light beams 21, 22, 23 and 24, and it is effective to supplement or improve the spectral characteristics of the dichroic mirrors 17 to 20. Although this is in 7th not shown, it is also effective to provide irradiation light such as excitation light around the light emitting point 1 to shine. When such irradiation light comes from a direction perpendicular to the optical axis of the condenser lens 2 is emitted without the condenser lens 2 can use the ratio of the irradiation light passing through the condenser lens 2 incident on the sensor can be reduced, which is advantageous in terms of sensitivity. Further, instead of the long pass filter 10, a dichroic mirror other than the dichroic mirrors 17 to 20 is arranged, and after the irradiation light is reflected by the above dichroic mirror, it becomes through the condenser lens 2 focused and on the light emitting point 1 blasted; and it is effective to adopt a so-called configuration for "incident light emission detection" using that from the light emitting point 1 emitted light through the condenser lens 2 bundled, transmitted by the dichroic mirror and then passed through a multicolor detection device such as that in 7th is recognized.

1. (1) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, in N Lichtstrahlen mit verschiedenen Wellenlängenanteilen aufgeteilt, die alle in der gleichen Richtung verlaufen.
2. (2) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, in N Lichtstrahlen mit verschiedenen Wellenlängenanteilen aufgeteilt, die alle in Richtung der optischen Achse der jeweiligen Kondensorlinse verlaufen.
3. (3) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, in verschiedene Wellenlängenanteile aufgeteilt, und die Richtung des aufgeteilten Lichts ist senkrecht zur Anordnungsrichtung der lichtemittierenden Punkteanordnung und der Kondensorlinsenanordnung.
4. (4) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, in verschiedene Wellenlängenanteile aufgeteilt, und die Richtung der Aufteilung ist senkrecht zur optischen Achse jeder Kondensorlinse.
5. (5) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, durch N dichroische Spiegel in verschiedene Wellenlängenanteile aufgeteilt, und die N dichroischen Spiegel sind in einer Richtung senkrecht zur Anordnungsrichtung der lichtemittierenden Punkteanordnung und der Kondensorlinsenanordnung angeordnet.
6. (6) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, durch N dichroische Spiegel in verschiedene Wellenlängenanteile aufgeteilt, und die N dichroischen Spiegel sind in einer Richtung senkrecht zur optischen Achse jeder Kondensorlinse angeordnet.
7. (7) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung werden M Lichtstrahlen, die erhalten werden, indem das von jedem lichtemittierenden Punkt emittierte Licht durch die Kondensorlinsenanordnung gebündelt wird, in N verschiedene Wellenlängenanteile aufgeteilt, um (MxN) Lichtstrahlen zu erhalten, die direkt auf den Sensor einfallen, ohne erneut gebündelt zu werden.
8. (8) Für M lichtemittierende Punkte der lichtemittierenden Punkteanordnung ist die optische Achse jeder Kondensorlinse der Kondensorlinsenanordnung zum Bündeln des von jedem lichtemittierenden Punkt emittierten Lichts senkrecht zur Sensorfläche angeordnet.
9. (9) N verschiedene Arten von dichroischen Spiegeln sind vorgesehen, und jeder der N verschiedenen Arten von dichroischen Spiegeln besteht aus einem Einzelelement. M Lichtstrahlen, die erhalten werden, indem das von M lichtemittierenden Punkten der lichtemittierenden Punkteanordnung emittierte Licht einzeln gebündelt wird, fallen parallel auf die jeweiligen dichroischen Spiegel ein.
10. (10) M Lichtstrahlen, die erhalten werden, indem das von M lichtemittierenden Punkten der lichtemittierenden Punkteanordnung emittierte Licht einzeln gebündelt wird, werden in N verschiedene Wellenlängenkomponenten aufgeteilt, um (MxN) Lichtstrahlen zu erhalten, die parallel auf den Einzelsensor einfallen.

In the in 7th The multicolor detection device shown here meet the diameter d of each light emitting point 1 , the distance p between the respective light emitting points 1 and between the respective condenser lenses 2 , the focal length f and the effective diameter D of each condenser lens 2 and the optical distance g between each condenser lens 2 and the sensor 30th any of the equations (3) to (7), the equations (8) to (12) and the equations (16) to (18), and therefore the predetermined high sensitivity and low crosstalk, and the size and cost, are achieved the detection device are reduced. The following features ( 1 ) to (10) summarize the features for reducing the size and cost of the multicolor detection device with the in 7th illustrated dichroic arrangement together. Not all of these characteristics have to be fulfilled, and it is already effective if some of them are fulfilled.

1. (1) For M light emitting points of the light emitting dot array, M light beams obtained by converging the light emitted from each light emitting point by the condenser lens array are divided into N light beams having different wavelength components, all of which travel in the same direction.
2. (2) For M light-emitting points of the light-emitting point array, M light beams obtained by converging the light emitted from each light-emitting point by the condenser lens assembly are divided into N light beams having different wavelength components, all in the direction of the optical axis of the respective condenser lens run away.
3. (3) For M light emitting points of the light emitting dot array, M light beams obtained by converging the light emitted from each light emitting point by the condenser lens array are divided into different wavelength components, and the direction of the split light is perpendicular to the arranging direction of the light emitting dot array and the condenser lens assembly.
4. (4) For M light emitting points of the light emitting point array, M light rays obtained by converging the light emitted from each light emitting point by the condenser lens array are divided into different wavelength components, and the direction of division is perpendicular to the optical axis of each condenser lens.
5. (5) For M light emitting points of the light emitting dot array, M light rays obtained by converging the light emitted from each light emitting point by the condenser lens array are divided into different wavelengths by N dichroic mirrors, and the N dichroic mirrors are in one direction arranged perpendicular to the direction of arrangement of the light-emitting point arrangement and the condenser lens arrangement.
6. (6) For M light emitting points of the light emitting dot array, M light rays obtained by converging the light emitted from each light emitting point by the condenser lens array are divided into different wavelengths by N dichroic mirrors, and the N dichroic mirrors are in one direction arranged perpendicular to the optical axis of each condenser lens.
7. (7) For M light emitting points of the light emitting point array, M light rays obtained by converging the light emitted from each light emitting point by the condenser lens array are divided into N different wavelength components to obtain (MxN) light rays that are directly incident on the Collapse sensor without being rebundled.
8. (8) For M light emitting points of the light emitting point array, the optical axis of each condenser lens of the condenser lens array for converging the light emitted from each light emitting point is arranged perpendicular to the sensor surface.
9. (9) N different types of dichroic mirrors are provided, and each of the N different types of dichroic mirrors is composed of a single element. M light rays obtained by individually converging the light emitted from M light emitting points of the light emitting point array are incident in parallel on the respective dichroic mirrors.
10. (10) M light beams obtained by individually converging the light emitted from M light emitting points of the light emitting dot array are divided into N different wavelength components to obtain (MxN) light beams incident in parallel on the single sensor.

The above relates to the case that the respective light beams obtained by converging the light emitted from the light emitting point array by respective condenser lenses of the condenser lens assembly are directly incident on the sensor without being re-bundled by a lens other than the condenser lens. The following describes a case in which the respective light beams obtained by converging the light emitted from the light emitting dot array by respective condenser lenses of the condenser lens assembly are directly incident on the sensor after being re-converged by a lens other than the condenser lenses. In the following, a lens for re-focusing each light beam is referred to as a “re-condenser lens”.

In the derivation of equations (1) to (18), the optical distance between the condenser lens and the sensor is defined as g, but when the re-condenser lens is used, equations (1) to (18) can be used as conditions for high sensitivity and low crosstalk can be used when g is the optical distance between the re-condenser lens and the sensor. That is, in 3 or 4th can change the position of the picture 7th of the light emitting point can be regarded as a position of the re-condenser lens, but it is not the position of the sensor. Because if the conditions for high sensitivity and low crosstalk are not met at the position of the re-condenser lens (e.g. when the overlap of the adjacent light beams is too great), the performance in terms of high sensitivity and low crosstalk will be behind the re-condenser lens irrespective of the configuration not improved. At the position of the optical distance g from the condenser lens, the same number of re-condenser lenses as condenser lenses are aligned at a distance p so that the optical axes of the paired condenser lenses and re-condenser lenses coincide. When “D” is the effective diameter of the re-condenser lens, which is the effective diameter of the condenser lens, the loss of the amount of detection light due to the re-condenser lens can be prevented, and a re-condenser lens assembly can be configured. The sensor is located at a position that is behind the re-condenser lens, and the optical path length from the condenser lens is greater than g. It can be seen from this that the conditions for realizing high sensitivity and low crosstalk in 3 or 4th are also given when using the re-condenser lens.

8th Fig. 3 is a schematic illustration of a multicolor detection device employing a wavelength dispersion element and a re-condenser lens assembly. 8 (a) Figure 13 is a schematic representation of the multicolor detection device viewed in a direction perpendicular to a plane defining each optical axis of the condenser lenses 2 contains, 8 (b) Fig. 3 is a schematic cross-sectional view of the multicolor detection device showing the optical axes of a condenser lens 2 and a paired re-condenser lens 33 and is perpendicular to the direction of arrangement of the condenser lens arrangement and the re-condenser lens arrangement, and 8 (c) shows one of a two-dimensional sensor 37 recognized image 42. Here is an example of the implementation of a three-color recognition.

As in 8 (a) is represented by the light emitting point 1 emitted light through the condenser lens 2 bundled and transmitted by the long-pass filter 10 as a light beam 9. These operations are the same as in 6th . Thereafter, the respective light rays 9 fall as in FIG 8 (b) shown, parallel to a common transmission diffraction grating 31 a, which is a wavelength dispersion element to be subjected to wavelength dispersion in a direction perpendicular to the direction of the light emitting dot array, by the re-condenser lens 33 to be bundled again and onto the two-dimensional sensor 37 to invade. Here, light rays 34, 35 and 36 show light rays with central wavelengths the A, B and C fluorescence. As in 8 (c) can be shown on the image 42 of the two-dimensional sensor 37 a wavelength dispersed image 41 des from each light emitting point 1 emitted light can be obtained. Here pictures 38, 39 and 40 show in the wavelength-dispersed image 41 displays images of the light rays with the central wavelengths of A, B and C fluorescence, respectively.

In the in 8th The multicolor detection device shown here meet the diameter d of each light emitting point 1 , the distance p between the respective light emitting points 1 and between the respective condenser lenses 2 , the focal length f and the effective diameter D of each condenser lens 2 and the optical path length g of each condenser lens 2 and any re-condenser lens 33 any of the equations (3) to (7), the equations (8) to (12) and the equations (16) to (18), and therefore the predetermined high sensitivity and low crosstalk, and the size and cost, are achieved the detection device are reduced.

8th shows that, when using the re-condenser lens, it is better to use one of equations (3) to (7), equations (8) to (12) and equations (16) for the optical distance g of the condenser lens and the re-condenser lens ) to (18). Accordingly, it is effective to satisfy any one of the equations (3) to (7), the equations (8) to (12), and the equations (16) to (18) by inserting the optical element that allows the respective light beams in to run in different directions, or the optical element for increasing the distance between the respective light beams is arranged at the position at which the optical distance from the condenser lens for converging the light emitted from the respective light-emitting points of the light-emitting point array by Forming rays of light is equal to g.

For example, in 9 a prism as the optical element 43 behind the long pass filter 10 in 6th arranged, the respective light beams 9 parallel to a common two-dimensional color sensor 11 come in after going in different directions and be recognized. 9 (a) Figure 10 shows the multicolor detection device in a direction perpendicular to a plane defining each optical axis of the condenser lenses 2 contains, and 9 (b) represents one of the two-dimensional color sensor 11 recognized image 12. The prism 43 changes the direction of refraction and a refraction angle according to the position of incidence of each light beam 9 and increases the distance between each light beam 44 after the refraction. Assume the optical distance between the condenser lens 2 and the prism 43 if g, then equations (1) to (18) directly become conditions for high sensitivity and low crosstalk. Because the distance between the pictures 45 of the light emitting points on the image 12 of the two-dimensional color sensor 11 compared to the case of 6th is increased, it is possible to see an increase in crosstalk behind the prism 43 to avoid and that from the respective light-emitting points 1 emitted light can be easily distinguished from one another. Because the distance between the pictures 45 of the light emitting points is large, it is not necessary that the sensor be a common two-dimensional color sensor as in FIG 9 and each light beam 44 can also be detected by a single color sensor. When the prism 43 on the configuration of 7th is applied, the prism can 43 be arranged between the long pass filter 10 and the dichroic arrangement. Because the optical path length between the condenser lens 2 and the prism 43 can be made smaller than the optical path length between the condenser lens 2 and the two-dimensional sensor 30th , the condition for low crosstalk is more easily met. Although the distance in the light beam dividing direction of the images 25 to 28 of the light emitting points in 7 (c) does not change, the crosstalk is slightly reduced as the distance in the direction of the light emitting dot array increases.

As described above, by sharing the dichroic mirror, the filter, the total reflection mirror, the diffraction grating and the sensor for the plurality of light emitting points, the configuration of the device is simplified and the implementation is facilitated. In addition, the overall size of the detection device is reduced. Further, according to the above configuration, the detection efficiency and the spectroscopic accuracy are the same for each light emitting point, and fluctuations in sensitivity and color discrimination can be reduced.

10 shows an example in which behind the long pass filter 10 in 6th an optical fiber 46 is arranged as the optical element. The light beam 9 is incident on incident ends of the respective optical fibers 46 and is emitted from their light-emitting ends with an increased spacing, and the emitted light beams 47 are incident in parallel on a common two-dimensional color sensor 11 one and are recognized. 10 (a) Figure 13 shows the multicolor detection device in a direction perpendicular to a plane defining each optical axis of the condenser lens 2 contains, and 10 (b) represents one of the two-dimensional color sensor 11 recognized image 12.

When the optical distance between the condenser lens 2 and the incident end of the optical fiber 46 is g, equations (1) to (18) directly become conditions for high sensitivity and low crosstalk. Because the distance between the pictures 48 of the light emitting points on the image 12 of the two-dimensional color sensor 11 compared to the case of 6th is increased, it is also possible to avoid an increase in crosstalk behind the optical fiber 46 and that from the respective light emitting points 1 emitted light can be easily distinguished from one another. Because the distance between the pictures 48 of the light emitting points is large, it is not necessary that the sensor be a common two-dimensional color sensor as in FIG 10 and each light beam 47 can also be detected by a single color sensor. When the optical fiber 46 is set to the configuration of 7th is applied, the incident end of the optical fiber 46 may be at the position of the two-dimensional sensor 30th in 7th be arranged, that is, behind the dichroic arrangement. Furthermore, the optical fiber 46 can all images 25 to 28 of the light emitting points in 7 (c) correspond. This makes it possible to arbitrarily set the spacing and arrangement of the images 25 to 28 of the light emitting points, and therefore it is easy to recognize the individual image with a single sensor or with a desired one-dimensional or two-dimensional sensor.

Embodiments of the present invention are described below.

[Embodiment 1]

11 Fig. 13 is a schematic diagram showing a configuration example of a DNA sequencer having a capillary arrangement. An analysis procedure is now on 11 Described with reference. First, the sample injection end 50 becomes a plurality of capillaries 49 (in 11 are four capillaries 49 shown) is immersed in a cathode-side buffer solution 60, and the sample rinsing device 51 is immersed via a polymer block 55 in an anode-side buffer solution 61. A valve 57 of the pump block 55 is closed, a polymer solution inside the pump block 55 is pressurized through a syringe 56 connected to the pump block 55, and the polymer solution is from the sample rinsing end 51 towards the sample injection end 50 into the respective capillaries 49 filled. Then, the valve 57 is opened, and various samples are fed into the respective capillaries from the sample injection ends 50 49 injected, whereupon with a power source 62 a high voltage is applied between a cathode electrode 58 and an anode electrode 59, whereby the capillary electrophoresis begins. DNAs marked with fluorescent substances in four colors migrate electrophoretically from the sample injection end 50 to the sample flush end 51.

Part of a coating on each capillary 49 at a position (laser irradiation position 52) a fixed distance from the sample injection end 50 where the DNAs in each capillary 49 migrate electrophoretically, is removed so that the laser irradiation positions 52 of the capillaries 49 lie on the same plane (arrangement plane) and are simultaneously irradiated by a laser beam 54 from a laser light source 53 is oscillated, focused by a lens and initiated along the plane of arrangement from one side of the plane of arrangement. The DNAs marked in four colors by the fluorescent substances migrate electrophoretically in each capillary 49 and vibrate and fluoresce as they pass through the laser irradiation position 52. Light emanating from the inside of each capillary 49 is emitted forms a light-emitting dot array, and each emitted light is emitted from a direction perpendicular to the plane of arrangement (direction perpendicular to a paper surface in 11 ) from the in 6th to 10 shown multicolor detection device detected.

In the present embodiment, a case will be described in detail in which the multicolor detection device having a dichroic arrangement for the in 11 capillary DNA sequencer shown is used. The laser irradiation positions 52 of the four capillaries 49 with an outer diameter of 0.36 mm and an inner diameter of 0.05 mm are arranged in the same plane with an interval of p = 1 mm, and the laser beam 54 narrowed to a diameter of 0.05 mm is irradiated on one side of an arrangement plane whereby a light emitting dot arrangement is obtained in which four (M = 4) of the light emitting dots 1 with an effective diameter of d = 0.05 mm are arranged at a distance of p = 1 mm. Here the effective diameter of the light-emitting point corresponds to the inner diameter of the capillary. Here, the wavelength of the laser beam 54 was set to 505 nm, and the fluorescence of four colors (maximum wavelength of light emission) were A fluorescence (540 nm), B fluorescence (570 nm), C fluorescence (600 nm ) and the D fluorescence (630 nm). The total width of the light emitting dot array was set to W = p * (M-1) = 3 mm. That from every light emitting point 1 emitted light is respectively from each condenser lens 2 a condenser lens assembly 8th bundled in which four condenser lenses 2 with a focal length of f = 1.5 mm and one effective diameter D = 1 mm are arranged at intervals of p = 1 mm. The above d, p, f, and D are substantially the same for the light emitting points and the condenser lenses, but do not necessarily have to be the same. In this case, d, p, f and D are average values of a plurality of light emitting points and the condenser lenses.

12th Figure 3 is a cross section of a multicolor detection device showing the optical axis of a condenser lens 2 contains, which is perpendicular to the direction of arrangement of the condenser lens arrangement 8th stands. But 12th differs from 7th in that the optical axis of each condenser lens 2 and a traveling direction of the light beam after being divided by the dichroic array is vertical, that is, the optical axis of each condenser lens 2 is parallel to a sensor surface of the two-dimensional sensor 30th . Otherwise it corresponds 7th . The light emitting point 1 is in 12th left out.

The long pass filter 10 with a width α = 5 mm, a thickness β = 1 mm and a depth γ = 5 mm is at a position at a distance of 3 mm from the condenser lens 2 arranged so that a normal to it and the optical axis of the condenser lens 2 are parallel to each other. Further, the dichroic mirrors 17, 18, 19 and 20 are formed with a multilayer or a single layer on a lower right front surface of a quartz substrate (refractive index n ₀ = 1.46) and are arranged at intervals of 5 mm so that their normal by 45 degrees to the optical axis of the condenser lens 2 is inclined. The quartz substrate has a width α = 5 mm, a thickness β = 1 mm and a depth γ = 5 mm. The dichroic mirror 20 can as in 7th be replaced by a total reflection mirror. 12th represents a side surface of α × β in both the long pass filter 10 and the dichroic mirrors 17 to 20, and γ is in a direction perpendicular to the paper surface. Furthermore, an anti-reflection layer is formed on a left upper front surface of the dichroic mirrors 17, 18 and 19 in order to reduce reflection loss. In addition, a light shielding layer is formed on all side surfaces of the dichroic mirrors 17, 18, 19 and 20 in order to prevent the penetration of light and to avoid undesired stray light. A right end of the dichroic mirror 17 is at a position spaced 5 mm from the condenser lens 2 (that is, at a position 2 mm from the long pass filter 10). The upper and lower ends of the dichroic mirrors 17, 18, 19 and 20 are each arranged in the same plane. The two-dimensional sensor 30th is arranged at a distance of 5 mm from the lower ends of the dichroic mirrors 17, 18, 19 and 20. All elements of the optical system are arranged in the air. The arrangement in air is done to improve the spectral performance of the dichroic mirror.

Eleven light beam elements 65 shown in 12th show that the above dichroic arrangement is designed to split incident parallel light beams into four light beams with a maximum width of the light beams, and represent the result of calculating their optical path from the laws of reflection and refraction the maximum width of the light beam is referred to as the opening 63 of the dichroic array, and a size thereof is expressed by W. The opening width represents the maximum width of the light beam at which the incident light beam can be divided in a suitable manner by the dichroic arrangement. The opening width W generally differs from a width d '(s) of the light beam obtained by focusing the point from the light emitting point 1 emitted light through the condenser lens 2 in the optical path length s of the condenser lens 2 is obtained. In addition, d '(s) is the real width of a light beam, and W represents a maximum value of the width of the light beam that can be accepted by the multicolor detection device under a given condition. That is, it is desirable that W≥d '(s) is set so that an amount of light passes through the condenser lens 2 bundled light beam is divided without loss of light quantity. In addition, it is desirable that W is larger because it reduces the tolerance of a deviation between a central axis of the through the condenser lens 2 bundled light beam and a central axis of the opening width 63 is increased compared to. The eleven light beam elements 65 extending from the condenser lens 2 spread to the left, are arranged so that they have the same spacing within the opening width 63 of the light beam and run parallel to one another.

As in 12th As shown, the respective light beam elements 65 are displaced in parallel upwards by internal refraction with each passage through the respective dichroic mirrors 17 to 19. To reduce this influence, the optical axis of the condenser lens are 2 and the center of the light beam element emitted from the condenser lens is located below a center of a lower right front surface of the dichroic mirror 17. The optical path length s, which is an optical distance between the condenser lens 2 and the two-dimensional sensor 30th is, however, is different in the four split light beams, as in 12th can be seen. The light beam which passes through the dichroic mirrors 17 to 19 and is reflected by the dichroic mirror 20 has the longest optical path 64. When there are multiple optical paths between the condenser lens and the sensor, the longest optical path length has been 64 below defined as the optical path length of the light detecting device, and the length thereof was expressed as g. In the multicolor detection device shown in 12th is shown, the opening width 63 was calculated as W = 2.1 mm, and the optical path length 64 as g = 29 mm. The above W and g are substantially the same for the light emitting points and the condenser lenses, but do not necessarily have to be the same. In this case, W and g are average values of a plurality of light emitting points and the condenser lenses.

An optical distance between the light emitting point 1 and the condenser lens 2 with a focal length of f = 1.5 mm is set to about 1.58 mm. This causes the from the light emitting point 1 emitted light at an optical distance g = 29 mm from the condenser lens 2 is shown, with an image magnification of m = 18.3 according to equation (1) and a diameter d '= 0.92 mm of the image 7th of the light emitting point 1 with a diameter of d = 0.05 mm according to equation (2).
Since the effective diameter D = 1 mm of the condenser lens is larger than the diameter d ', the optical path length s from the condenser lens is set as d' (s) ≤ 1 mm, and d '(s) W = 2.1 mm , based on is 0 mm≤s≤29 mm. Thereby, the light beam, which is split into four light beams by the dichroic mirrors 17 to 20, can use the two-dimensional sensor 30th Achieve lossless.

In the multicolor detection apparatus described above, equations (4), (9) and (18) are satisfied, and it was found that a relative detection light amount larger than 100%, an accurate relative detection light amount larger than 100%, high sensitivity with 0% crosstalk and the low crosstalk condition can be achieved. A size of the multicolor detection device is determined by the total width AW = 3 mm of the light emitting dot array and can be, as shown in FIG 12th shown to be smaller than a volume (668 mm ² ) of a rectangular parallelepiped having a width of 24.2 mm in the direction of an optical axis of the condenser lens 2 and a width of 9.2 mm in a direction perpendicular to the optical axis of the condenser lens 2 and the light emitting dot array. That is, compared with the case disclosed in PTL 1, a total size of the fluorescence detection device can be reduced to 1 / 2,400 times. Since all the optical elements used are fine, great cost savings are possible. The above m and d 'are essentially the same for the light emitting points and the condenser lenses, but do not necessarily have to be the same. In this case, m and d 'are average values of a plurality of light emitting points and the condenser lenses.

[Embodiment 2]

In the multicolor detection device of embodiment 1 , in the 12th is shown, the arrangement directions of the dichroic arrangement, the condenser lens arrangement and the light-emitting point arrangement are all parallel to the sensor surface of the two-dimensional sensor 30th , so there is no interference in between. Since the planes of arrangement of the respective capillaries 49 the capillary arrangement perpendicular to the sensor surface of the two-dimensional sensor 30th this device configuration has a disadvantage that interference may occur therebetween. The present embodiment therefore provides a multicolor detection device in order to solve the above disadvantage.

In the capillary DNA sequencer, the configurations from the capillary array to the condenser lens array are the same as in the embodiment 1 , and the configuration of 12th is made possible by a configuration of 13th replaced. 13th Fig. 13 is a cross-sectional diagram schematically showing a multicolor detection device. The cross-sectional diagram includes an optical axis of a condenser lens 2 , and its paper plane is perpendicular to the direction of arrangement of the condenser lens arrangement 8th , and the optical axis of each condenser lens 2 is parallel to a traveling direction of the light beam after division, that is, the optical axis of each condenser lens 2 and the sensor area of the two-dimensional sensor 30th stand perpendicular to each other what 7th corresponds to. The long pass filter 10 having a width α = 5 mm, a thickness β = 1 mm and a depth γ = 5 mm is at a position at a distance of 3 mm from the condenser lens 2 arranged so that a normal to it and the optical axis of the condenser lens 2 are parallel to each other. Further, the dichroic mirror 17 is formed with a multilayer film on an upper front surface of a quartz substrate (refractive index n ₀ = 1.46) with a width α = 5 mm, a thickness β = 1 mm and a depth γ = 5 mm, and the dichroic mirrors 18, 19 and 20 with the multilayer or single layer formed on a lower right front surface thereof are arranged at intervals of 5 mm so that their normal to the optical axis of the condenser lens 2 is inclined at 45 degrees at 5 mm intervals. In addition, a light shielding layer is formed on all side surfaces of the dichroic mirrors 17, 18, 19 and 20 in order to prevent the penetration of light and to avoid undesired stray light.

An upper end of the dichroic mirror 17 is at a distance of 5 mm from the condenser lens 2 arranged. The upper and lower ends of the dichroic mirrors 17, 18, 19 and 20 are up, respectively arranged on the same level. The two-dimensional sensor 30th is arranged at a distance of 5 mm from the lower ends of the dichroic mirrors 17, 18, 19 and 20. As in 12th eleven light beam elements 65 are shown. As a result, in the multicolor detection device disclosed in 13th is shown, the aperture width 63 is calculated as W = 1.7 mm, and the optical path length 64 as g = 28 mm, whereby the same performance as in 12th can be reached.

An optical distance between the light emitting point 1 and the condenser lens 2 is set to about 1.58 mm. This causes the from the light emitting point 1 emitted light at an optical distance g = 28 mm from the condenser lens 2 with an image magnification of m = 17.7 and a diameter of the image 7th of the light emitting point of d '= 0.88 mm. Since the effective diameter D = 1 mm of the condenser lens is larger than the diameter d ', the optical distance s from the condenser lens becomes 2 set as d '(s) ≤1mm, and d' (s) ≤W = 1.7mm with respect to 0mm≤s≤28mm. Thereby, the light beam, which is split into four light beams by the dichroic mirrors 17 to 20, can use the two-dimensional sensor 30th Achieve lossless.

In the multicolor detection apparatus described above, equations (4), (9) and (18) are satisfied, and it was found that a relative detection light amount larger than 100%, an accurate relative detection light amount larger than 100%, high sensitivity with 0% crosstalk and the low crosstalk condition can be achieved. A size of the multicolor detection device is determined by the total width AW = 3 mm of the light emitting dot array and can be, as shown in FIG 13th shown to be smaller than a volume (818 mm ² ) of a rectangular parallelepiped having a width of 14.2 mm in the direction of an optical axis of the condenser lens 2 and a width of 19.2 mm in a direction perpendicular to the optical axis of the condenser lens 2 and the light emitting dot array. That is, compared with the case disclosed in PTL 1, a total size of the fluorescence detection device can be reduced to 1 / 2,000 times. Since all the optical elements used are fine, great cost savings are possible.

Since in the multicolor detection device disclosed in 13th the arrangement directions of the dichroic arrangement, the condenser lens arrangement, the light-emitting point arrangement and the arrangement plane of the capillary arrangement are all parallel to the sensor surface of the two-dimensional sensor 30th there is no interference between them, so that their implementation becomes easy.

[Embodiment 3]

The inside diameter of each capillary 49 , that is, the diameter of the light emitting point 1 was in the embodiments 1 and 2 set to d = 0.05 mm. Here an enlargement of this condition to d = 0.075 mm was examined. Using the in 13th The multicolor detection device shown was that of the light emitting point 1 emitted light mapped so that an image magnification to m = 17.7 and the diameter of the image 7th of the light emitting point was set to d '= 1.33 mm at an optical distance g = 28 mm. Since the effective diameter D = 1 mm of the condenser lens is larger than the diameter d ', the optical path length is s from the condenser lens 2 set as d '(s) ≤1.33mm, and d' (s) ≤W = 1.7mm with respect to 0mm≤s≤28mm. Thereby, the light beam which is split into four light beams by the dichroic mirrors 17 to 20, as shown in FIG 13th the two-dimensional sensor 30th Achieve lossless.

It was found that equations (4) and (9) are satisfied in the above multicolor detection device in the same manner as in FIG 13th . However, equation (18) is not satisfied and equation (17) is satisfied. Accordingly, a relative detection light amount greater than 100%, an accurate relative detection light amount greater than 100%, high sensitivity with 25% or less crosstalk, and low crosstalk can be achieved, from which it can be seen that the crosstalk is worse than in FIG 13th . In the present embodiment, the crosstalk can be further reduced by shortening the optical path length g, thereby increasing the image magnification m and the diameter d 'of the image 7th be reduced. In order to shorten the optical path length g, it is considered effective to reduce the size and the spacing of the respective dichroic mirrors 17 to 20.

Referring to 14th , has been made regarding the long pass filter 10 and the dichroic mirrors 17 to 20 in the conditions of 13th the width α, the thickness β and the depth γ are reduced from α = 5 mm, β = 1 mm and γ = 5 mm to α = 2.5 mm, β = 1 mm and γ = 5 mm. The spacing between the respective dichroic mirrors 17 to 20 was reduced from x = 5 mm to x = 2.5 mm.

In this case, the position of a left end of the dichroic mirror 17 is correct in a lateral direction (direction perpendicular to the optical axis of the condenser lens 2 ) in 14th coincides with the position of a right end of the dichroic mirror 18. Similarly, a position of a left end of the dichroic mirror 18 in a lateral direction coincides with a position of a right end of the dichroic mirror 19 and a position of a left end of the dichroic mirror 19 in a lateral direction corresponds to a position of a right end of the dichroic mirror 20 match. Otherwise the conditions are the same as in 13th . 14th is drawn to the same scale as 13th .

As a result, the optical path length 64 can be shortened from g = 28 mm to g = 19 mm. Accordingly, the image magnification according to equation (1) becomes m = 11.7 and the diameter of the image of the light emitting point according to equation (2) becomes d '= 0.88 mm, and equations (4), (9) and ( 18) are met. That is, the relative detection light amount greater than 100%, the precise relative detection light amount greater than 100%, high sensitivity with 0% crosstalk, and low crosstalk can be achieved. It can be seen, however, that the opening width 63 has been greatly reduced from B = 1.7 mm to W = 0.03 mm. That is, it has been found to be there for that through the condenser lens 2 bundled emitted light is impossible to use the two-dimensional sensor 30th to achieve without loss. Specifically, since a relative ratio of the thickness β1 = mm of each dichroic mirror increases as the size of each dichroic mirror decreases and the spacing between them decreases, an influence of the light rays which are parallel-displaced upward by internal refraction each time the dichroic mirrors pass through cannot be ignored become.

In order to overcome this influence, the arrangement of the dichroic mirrors 17 to 20 is therefore changed from an arrangement on the same plane to a step-like arrangement as in FIG 15th shown. That is, the relative positions of the dichroic mirrors 17 to 20 in the direction of the optical axis of the condenser lens 2 were modified according to the parallel displacement from the arrangement on the same plane. The following is the difference compared to 14th described. First, the upper end of the dichroic mirror 17 is placed at a position 6.3 mm from the condenser lens 2 arranged. Then, the lower end of the dichroic mirror 18 is displaced to a side opposite to a direction of propagation of a light beam passing through the dichroic mirror 17, that is, by y = 0.7 mm in comparison with the lower end of the dichroic mirror 17 15th . Next, the lower end of the dichroic mirror 19 is displaced to a side opposite to a traveling direction of a light beam reflected by the dichroic mirror 18, that is, by z = 0.3, compared to the lower end of the dichroic mirror 18 mm in 15th . Finally, the lower end of the dichroic mirror 20 is displaced to a side opposite to a direction of propagation of a light beam reflected by the dichroic mirror 19, that is, by z = 0.3 mm, compared to the lower end of the dichroic mirror 19 in 15th .

As a result, it was found that the opening width 63 of W = 0.03 mm in 14th could be expanded to W = 1.3 mm. On the other hand, since the optical path length is 64 compared to 14th has increased slightly to g = 21 mm, equation (1) gives an image magnification of m = 13 and equation (2) a diameter of the image 7th of the light emitting point of d '= 0.98 mm. Since the effective diameter D = 1 mm of the condenser lens is larger than d ', the optical path length s from the condenser lens is set to d' (s) 1 mm, and d '(s) W = 1.3 mm in relation to 0 mm≤s≤21 mm. Thereby, the light beam, which is split into four light beams by the dichroic mirrors 17 to 20, can use the two-dimensional sensor 30th Achieve lossless.

In the multicolor detection device described above, equations (4), (9) and (18) are expressed in the same manner as in FIG 14th is satisfied, and it has been found that a relative detection light amount greater than 100%, an accurate relative detection light amount greater than 100%, high sensitivity with 0% crosstalk, and the low crosstalk condition can be achieved. A size of the multicolor detection device is determined by the total width AW = 3 mm of the light emitting dot array and can be, as shown in FIG 15th shown to be smaller than a volume (414 mm ² ) of a rectangular parallelepiped having a width of 13.8 mm in the direction of an optical axis of the condenser lens 2 and a width of 10 mm in a vertical direction to the optical axis of the condenser lens 2 and the light emitting dot array. That is, compared with the case disclosed in PTL 1, a total size of the fluorescence detection device can be reduced to 1/3,900 times. Since all the optical elements used are fine, great cost savings are possible.

[Embodiment 4]

As is apparent from the above-described embodiments, it is important that the multicolor detection device with the dichroic arrangement achieve an enlargement of the opening width W and a shortening of the optical path length g, which are in conflict with each other. However, due to the miniaturization of the device, it was difficult to reconcile the two. The adaptation of the arrangement distance x of the dichroic arrangement and the step-like offset y and z, which in embodiment 3 are effective ways to solve the problem described above. In embodiment 4th the arrangement distance and the staggered arrangement were generalized, and a general solution was derived in order to reconcile the enlargement of the opening width W and the shortening of the optical path length g. An essential feature of the dichroic mirror arrangement in the present invention is that the thickness β of the dichroic mirror is taken into account. In the prior art, the size of a dichroic mirror (α and γ) is large enough. Therefore, the thickness β does not have to be considered. However, when they are miniaturized, it is important to arrange them in consideration of the thickness β. In particular, if each dichroic mirror is not installed in the air but in a glass material, there is nothing to prevent the assumption of a thickness β of β = 0.

16 Fig. 13 is a cross-sectional diagram schematically showing the multicolor detection device. The cross-sectional diagram includes an optical axis of a condenser lens 2 which are perpendicular to the direction of arrangement of the condenser lens arrangement 8th stands. Further provides 16 represents a configuration of a multicolor detection device when, for a given width α and thickness β of the dichroic mirror, the opening width W has been maximized and the optical path length g has been minimized. The condenser lens 2 , the long pass filter 10 and the two-dimensional sensor 30th were in 15th left out.

As in 16 shown, a light beam 70 incident from top to bottom with the opening width W repeats the reflection and the passage through dichroic mirrors M (1), M (2), M (3) ... and M (N) in order to generate light beams F ( 1), F (2), F (3) ... and F (N), which are emitted from top to bottom. The number of dichroic mirrors, that is, the division number of the light beam 70 is in the example of FIG 16 four, but in the present embodiment the number has been generalized as N (N≥2). The Nth dichroic mirror M (N) can also be replaced by a total reflection mirror. Above, as in 12th to 15th As shown, the optical path length g of the light emitting detection device was taken as the optical path length of the optical path from the condenser lens 2 to the sensor 30th defined with the maximum length. In the following, as in 16 shown, an optical path length L of the dichroic mirror assembly as part of the optical path length g in the optical path with the longest length (the optical path from light beam 70 to light beam F (N)) as an optical path length from a position at the same level as the top end (the uppermost end of the dichroic mirror M (N)) of the dichroic mirror arrangement on the optical axis of the light beam 70 to a position at the same level as the lowermost end (the lowermost end of the dichroic mirror M (1)) of the dichroic mirror arrangement on a optical axis of the light beam F (N) is defined.

Each dichroic mirror has an optical coating with a refractive index no on at least one front surface of a transparent substrate and is arranged at a distance x in air. For every dichroic mirror in 16 is defined a normal vector from a left front surface in a left upper direction. Each dichroic mirror is set in such a way that each normal vector is opposite to a direction opposite to the direction of propagation of the light beam 70 (direction from bottom to top in 16 ) is inclined by an angle θ ₀ ( _{0 0} 90 °). 16 illustrates θ ₀ = 45 °, but in the present embodiment, θ ₀ can be set to any angle of 0 ° ₀ 90 °. As described above, the dichroic mirrors M (1) to M (N) are arranged approximately parallel to one another at approximately equal intervals. Furthermore, the lower end of the dichroic mirror M (2) is arranged such that it is offset upward by y with respect to the lower end of the dichroic mirror M (1), that is, by y in a direction opposite to the direction of propagation of the light beam F (1). In addition, the lower end of the dichroic mirror M (3) is arranged such that it is offset upward by z with respect to the lower end of the dichroic mirror M (2), that is, by z in a direction opposite to the direction of propagation of the light beam F (2). Accordingly, on the basis of 3 n N, the lower end of the dichroic mirror M (n) is arranged to be offset upward by z with respect to the lower end of the dichroic mirror M (n-1), that is , by z in a direction opposite to the direction of propagation of the light beam F (n-1).

Here, the angle of incidence of the light beam on an incidence surface of the dichroic mirror M (1) is θ ₀ , and the angle of refraction θ _{1 of} the light beam on the incidence surface (upper left front surface) of the dichroic mirror M (1) is as follows: $${\mathrm{<figure-callout\; id="1"\; label="\theta "\; filenames="DE112016007594B3\_0060.png,DE112016007594B3\_0062.png"\; state="\{\{state\}\}">\theta </figure-callout>}}_1={\text{sin}}^{-1}\left(1/{\text{<figure-callout id="0" label="n" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">n</figure-callout>}}_0*\text{sin}{\mathrm{\theta }}_0\right)$$

In addition, the angle of incidence of the light beam on an incidence surface (lower right front surface) of the dichroic mirrors M (2) to M (N) is 90 ° -θ ₀ , and the angle of refraction θ _{2 of} the light beam on the incidence surface of the dichroic mirrors M (2) to M (N) is as follows: $${\mathrm{<figure-callout\; id="2"\; label="\theta "\; filenames="DE112016007594B3\_0062.png,DE112016007594B3\_0065.png"\; state="\{\{state\}\}">\theta </figure-callout>}}_2={\text{sin}}^{-1}\left(1/{\text{<figure-callout id="0" label="n" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">n</figure-callout>}}_0*\text{sin}\left(90°-{\mathrm{\theta }}_0\right)\right)$$

The right end of the light beam 70 is shown as the right light beam end 66 with a dashed line, and a left end thereof is shown as the left light beam end 67 with a dot-dash line. The dashed line and the chain line represent the right end and the left end of the light rays F (1), F (2), F (3), ... and F (N).

In the following, two conditions for the best mode of execution for maximizing the opening width W and minimizing the optical path length L are referred to 16 described. The first condition is that the right end 66 of the light beam meets the left end corner 69 of the dichroic mirrors M (1), M (2), ... and M (N-1) shown in 16 is shown as triangular marks, or passes through their neighborhood. The second condition is that the left end 67 of the light beam must meet the lower end corner 68 of the dichroic mirror M (1) and the left end corner 69 of the dichroic mirror M (2), ... and M (N-1) shown in FIG 16 is shown as circular marks, or passes through their neighborhood. According to the above conditions, the geometric relationship in 16 derived the following relational equation.
First of all, the distance x between the dichroic mirrors M (1) to M (N) in the best execution mode is as follows: $$\text{X}={\text{x}}_0=\text{cos}{\mathrm{\theta }}_0*\mathrm{\alpha }+\text{<figure-callout id="0" label="sin θ" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">sin</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_0*\mathrm{\beta }$$

The opening width W in the best execution mode is as follows: $$\text{W.}={\text{W.}}_0={\text{a}}_{\text{w}}*\mathrm{\alpha }+{\text{b}}_{\text{w}}*\mathrm{\beta }$$

$${\text{b}}_{\text{w}}=-\text{cos}{\mathrm{\theta }}_0*\text{tan}{\mathrm{\theta }}_1$$

Where: $${\text{a}}_{\text{w}}=\text{cos}{\mathrm{\theta }}_0$$

$${\text{b}}_{\text{w}}=-\text{cos}{\mathrm{\theta }}_0*\text{<figure-callout id="1" label="tan θ" filenames="DE112016007594B3\_0060.png,DE112016007594B3\_0062.png" state="{{state}}">tan</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_1$$

Furthermore, in the best mode, the optical path length L is as follows: $$\text{L.}={\text{L.}}_0={\text{a}}_{\text{L.}}*\mathrm{\alpha }+{\text{b}}_{\text{L.}}*\mathrm{\beta }$$

$${\text{b}}_{\text{L}}\equiv \left(\text{N}-2\right)/\text{cos}{\mathrm{\theta }}_0*\left(2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)+1-\text{sin}\left({\mathrm{\theta }}_0+{\mathrm{\theta }}_2\right)\right)+\left(\text{N}-2\right)*\text{sin}{\mathrm{\theta }}_0+2*\text{cos}{\mathrm{\theta }}_0$$

Where: $${\text{a}}_{\text{L.}}\equiv \left(\text{N}-1\right)*\text{cos}{\mathrm{\theta }}_0+\text{<figure-callout id="0" label="sin θ" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">sin</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_0$$

$${\text{b}}_{\text{L.}}\equiv \left(\text{N}-2\right)/\text{cos}{\mathrm{\theta }}_0*\left(2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)+1-\text{sin}\left({\mathrm{<figure-callout\; id="0"\; label="\theta "\; filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png"\; state="\{\{state\}\}">\theta </figure-callout>}}_0+{\mathrm{\theta }}_2\right)\right)+\left(\text{N}-2\right)*\text{<figure-callout id="0" label="sin θ" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">sin</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_0+2*\text{cos}{\mathrm{\theta }}_0$$

$$\text{z}={\text{z}}_0=\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }$$

The step-like offsets y and z of each dichroic mirror M (1) through M (N) in the best mode are as follows: $$\text{Y}={\text{<figure-callout id="0" label="y" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">y</figure-callout>}}_0=\text{cos}{\mathrm{\theta }}_0*\mathrm{\beta }$$

$$\text{z}={\text{<figure-callout id="0" label="z" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">z</figure-callout>}}_0=\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }$$

As described above, x ₀ , W ₀ , L ₀ , y _0, and z _{0 are} all correlated with α and β.

The above α, β, n ₀ , θ ₀ , x and z are essentially the same among the dichroic mirrors and the total reflection mirror, but they do not necessarily have to be the same. In this case, α, β, n ₀ , θ ₀ , x and z are mean values of a plurality of dichroic mirrors.

By inversely solving the above, α, β and x can be derived to obtain a minimum value Wmin of a target opening width. From W ₀ ≥W _min and equation (22) results (here best execution mode in the case of an equal sign): $$\mathrm{\alpha }\ge -{\text{b}}_{\text{w}}/{\text{a}}_{\text{w}}*\mathrm{\beta }+1/{\text{a}}_{\text{w}}*{\text{W.}}_{\text{min}}$$

According to equation (21) results (here best execution mode in the case of an equal sign): $$\text{x}\ge \left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{w}}/{\text{a}}_{\text{w}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta }+1/{\text{a}}_{\text{w}}*\text{cos}{\mathrm{\theta }}_0*{\text{W.}}_{\text{min}}$$

α, β and x for obtaining a maximum value L _{max of} an aimed optical path length can be derived in the same way. According to equation (25) results (here best execution mode in the case of an equal sign): $$\mathrm{\alpha }\le -{\text{b}}_{\text{L.}}/{\text{a}}_{\text{L.}}*\mathrm{\beta }+1/{\text{a}}_{\text{L.}}*{\text{L.}}_{\text{Max}}$$

Equation (21) gives (here the best mode in the case of an equal sign). $$\text{x}\le \left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{L.}}/{\text{a}}_{\text{L.}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta }+1/{\text{a}}_{\text{L.}}*\text{cos}{\mathrm{\theta }}_0*{\text{L.}}_{\text{Max}}$$

W _min and L _max are substantially the same among the light emitting points and the condenser lenses, but do not necessarily have to be the same. In this case, W _min and L _{max are} average values of the plurality of light emitting points and condenser lenses.

For example, for a case with N = 4, n ₀ = 1.46 and θ ₀ = 45 °, FIG. 13 shows a range in which equations (31) and (33) are satisfied, with a horizontal axis β and a vertical axis x. The parameters are set to W _min = 0.5, 1, 2, 3 and 4 mm, L _max = 5, 10, 20, 30 and 40 mm. ↑ indicates an area above a straight line, and ↓ indicates an area below a straight line. For example, _{to achieve W min} = 0.5 mm and L _max = 20 mm, ↓ and x in 17th in an area above the straight line for ↑ W _min = 0.5 and an area below the straight line for ↓ L _max = 20.

If, for a diameter d of a given light-emitting point and a distance p of a given light-emitting point arrangement, the focal length f of the condenser lens and the optical path length g of the condenser lens and the sensor are chosen so that the conditions for high sensitivity in one of the equations (3) to (7) or (8) to (12) are satisfied, and when the equations (31) and (33) _{are satisfied by W min} = d 'and L _max = g, a compact multicolor detection device can be obtained with a dichroic arrangement configured which has high sensitivity. Here, d '= (gf) / f * d is expressed according to equation (2). Accordingly, if a focal length f of the condenser lens and an optical path length g of the condenser lens and the sensor are chosen for a diameter d of a given light-emitting point and a distance p of a given light-emitting point arrangement, both the conditions for low crosstalk of one of the equations (6) to (18) and equations (31) and (33) _{are satisfied by W min} = d 'and L _max = g, the dichroic arrangement can provide a compact multicolor detection device which has little crosstalk. In addition, when a focal length f of the condenser lens and an optical path length g of the condenser lens and the sensor are selected, both of which satisfy the conditions for high sensitivity of one of the equations (3) to (7) or (8) to (12) and the conditions satisfy one of the equations (6) to (18) for a low crosstalk, and the equations (31) and (33) can be satisfied by Wmin = d 'and L _max = g, with the dichroic arrangement, the compact multicolor detection device can be provided, which has a high sensitivity and a low crosstalk.

For example, if for d = 0.05 mm, p = 1 mm, f = 1.5 mm, D = 1 mm, g = 29 mm in embodiment 1, N = 4, n ₀ = 1.46, θ ₀ = 45 °, β = 1 mm, x = 5 mm is set, all of equations (4), (9), (18), (31), (31) and (33) are satisfied and a compact multicolor detection device becomes obtained with a dichroic arrangement that has high sensitivity and low crosstalk.

Next, the distance x between the dichroic mirrors M (1) to M (N) will be explained in detail. In the best mode of implementation, it may be desirable _{to adjust x 0} in equation (21) as described above. The following describes in detail how much deviation from the best mode of execution is allowed to get the effect. A solid line in 18th is the result of calculating a relationship between the distance x and the opening width W of the dichroic mirrors 17 and 18 in FIG 15th . In general, there is a possibility that the total opening width W becomes smaller than the above result as the total number N of dichroic mirrors increases. In this case, however, N = 2 is rated as an indicator. 15th represents a condition x = x ₀ = 2.5 mm, which was calculated using equation (21) for θ ₀ = 45 ° and β = 1 mm. In this case, the opening width is maximal at W = 1.3 mm, as in 18th shown. For x <x ₀ , W increases proportionally | xx ₀ | from, and at x = 1.6 mm, W = 0 mm. If x> x ₀ , however, W is W = 1.3 mm and constant. Dashed line in 18th represents a relationship between the distance x and a change ΔL in the optical path length L in 15th For x = x ₀ = 2.5 mm, ΔL = 0 mm was set here and displayed at the same height as W = 1.3 mm. Further, scales of a vertical axis of W (left side) and a vertical axis (right side) of ΔL were juxtaposed, and the vertical axis of ΔL was vertically inverted. In general, there is a possibility that ΔL becomes larger than the above result as the total number N of dichroic mirrors increases, but in this case N = 2 was evaluated as an indicator. It is also understood that ΔL increases proportionally to x.

Out 18th it can be seen that a rate of increase of W versus x in 1.6 mm x 2.5 mm and a rate of increase of ΔL versus x in 2.5 min x have approximately the same slope, and the slope is about 1. In other words, it can be seen that the power is proportional to | x - x ₀ | gets worse. In contrast, β is not taken into account in the prior art and corresponds to β = 0 mm. At this time, if the distance x ₀ in equation (21) is set to x = 1.8 mm in the case of an equivalent arrangement, the result is 18th corresponding to W = 0.4 mm. From the above, it can be seen that 1.8 mm x 3.2 mm is desirable in order to obtain performance equal to or better than the prior art. In general, the arrangement distance x between the dichroic mirror M (n) and M (n-1) shown in FIG 16 is specified as 2≤n≤N, expressed as follows: $$\text{cos}{\mathrm{\theta }}_0*\mathrm{\alpha }\le \text{cos}{\mathrm{\theta }}_0*\mathrm{\beta }+2*\text{<figure-callout id="0" label="sin θ" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">sin</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_0*\mathrm{\beta }$$

As a result, the opening width W can be enlarged and the optical path length L can be reduced.

Next, the step-like displacements y and z between the dichroic mirrors M (1) to M (N) will be explained in detail. As described above, in the best mode of execution it may be desirable to set Yo and Zo in equations (28) and (29), but the following examines in detail to what extent it is permissible to use the best mode of execution to have the effect of the to get staggered arrangement.

19 (a) FIG. 13 is a result of calculating a relationship between the stepped offset y and the opening width W of the dichroic mirrors 17 and 18 in FIG 15th . In general, there is a possibility that the total aperture W becomes smaller than the above result as the total number N of dichroic mirrors increases, but in this case, N = 2 was evaluated as an indicator. 15th represents a condition y = y ₀ = 0.7 mm calculated for θ ₀ = 45 ° and β = 1 mm from equation (28), but in this case as in 19 (a) shown, the opening width is maximum at W = 1.3 mm. In addition, W increases proportionally | yy ₀ | from. At y = 0 mm and 1.4 mm, W becomes W = 0.6 mm, and at y = -0.7 mm and 2.1 mm, W becomes W = 0 mm. Here a negative y indicates the step-like offset in an opposite direction to 15th at. Accordingly, it was found that the effect of the step-like displacement is achieved by setting 0 mm y y 1.4 mm.

Accordingly is 19 (a) the result of calculating a relationship between a stepped offset z and the opening width W of the dichroic mirrors 18 and 19 in 15th . 15th represents a condition z = z ₀ = 0.3 mm, which was calculated for θ ₀ = 45 ° and β = 1 mm from equation (29), but in this case as in 19 (b) shown, the opening width is maximum at W = 1.3 mm. also W increases proportionally | zz ₀ | from. At z = 0 mm and 0.6 mm, W becomes W = 1 mm, and at y = -1.1 and 1.7 mm, W = 0 mm. A negative z here shows the step-like offset in an opposite direction 15th at. Accordingly, it was found that the effect of the stepped offset is achieved by setting z to 0 mm z 0.6 mm. The above is generalized as described below. In 16 an end of a propagation side of a split light beam of the dichroic mirror M (2) opposite to the end of the propagation side of the split light beam of the dichroic mirror M (1) is displaced by y opposite to the propagation direction of the split light beam, where y is set as follows: $$0\le \text{y}\le 2*\text{cos}{\mathrm{\theta }}_0*\mathrm{\beta }$$

As a result, the opening width W can be enlarged and the optical path length L can be reduced.

Further, in the case of 3≤n≤N, the end of the propagation side of the split light beam of the dichroic mirror M (n) is offset from the end of the propagation side of the split light beam of the dichroic mirror M (n-1) by z opposite to the propagation direction of the split light beam , where z is set as follows: $$0\le \text{z}\le 2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }$$

As a result, the opening width W can be enlarged and the optical path length L can be reduced.

Set as described above 15th and 16 represents the configuration in which the optical axis of the condenser lens and the traveling direction of the split light beam are parallel to each other. On the other hand, when the optical axis of the condenser lens and the direction of propagation of the split light beam are vertical, as in FIG 12th If 2≤n≤N, the end of the propagation side of the split light beam of the dichroic mirror M (n) is offset by z opposite to the propagation direction of the split light beam compared to the end of the propagation side of the split light beam of the dichroic mirror M (n-1). As a result, the opening width W can be enlarged and the optical path length L can be reduced.

[Embodiment 5]

In embodiments 3 and 4, the opening width W of the dichroic arrangement is enlarged and the optical path length L is shortened by arranging a large number of dichroic mirrors in a step-like manner. In the present embodiment, a method is proposed to increase the opening width W and shorten the optical path length L without staggered arrangement, that is, when a plurality of dichroic mirrors are arranged on the same plane, that is, when the ends of the Propagation side of the split light beam of the respective dichroic mirror lie on the same plane.

14th of Embodiment 3 represents the result at θ ₀ = 45 ° while 20th represents the result at θ ₀ = 50 °. The other conditions are at 14th and 20th same, and in 14th and 20th the dichroic mirrors 17 to 20 are arranged on the same plane in both cases. Nevertheless, it was found that the opening width in 14th is only W = 0.03 mm, while the opening width in 20th is increased in a significant way to W = 0.9 mm. Here is the maximum optical path length between 14th and 20th not changed and is set to L = 19 mm. Accordingly, the image magnification is similar to that in 14th m = 11.7, and the diameter of the light emitting point image is d '= 0.88 mm, and equation (18) and equations (4) and (9) are satisfied. Therefore, the relative detection light amount is greater than 100%, the accurate relative detection light amount is greater than 100%, high sensitivity with 0% crosstalk, and the low crosstalk condition are achieved.

The following describes the reason why the above effects are achieved. As in 14th shown, the light beam in the case of θ ₀ = 45 ° in the space between the various dichroic mirrors propagates in a horizontal direction to the left, while inside each dichroic mirror it propagates in a left upper direction, whereby the light beam with each pass a dichroic mirror is gradually moved upwards, which limits the opening width W. In the case of θ ₀ = 50 ° ≥45 ° in 20th on the other hand, the light beam spreads in the space between the different dichroic mirrors in a lower left direction. Since the light beam propagates inside the respective dichroic mirror in a left upper direction, both directions are compensated and the vertical displacement of the light beam with each pass through a dichroic mirror is suppressed, whereby the Opening width W is increased. Therefore, it may be desirable to set θ ₀ greater than 45 °, there being an optimum value for maximizing the opening width W.

21 FIG. 10 shows a result of calculation of W when θ _{0 is} among those in 14th and 20th conditions shown is changed. It was found that W increases from θ ₀ = 45 °, reaches a maximum value of 0.92 mm _{at θ 0} = 52 ° and then drops to a value of approximately zero _{at θ 0 = 57 °.} That is, it has been found that W _{can be increased at 45 ° 0} 57 57 °.

The following describes a generalization with respect to the above. The following equations were made in the same manner as in 16 from a geometric relationship in 20th derived. The angle of refraction θ _{1 of} the light beam on the incidence surface of the dichroic mirror M (1) is as in equation (19), and the angle of refraction θ _{2 of} the light beam on the incidence surface of the dichroic mirrors M (2) to M (N-1) is as in equation (20). A downward offset S ↓ of the light beam, which propagates in the space between different dichroic mirrors in a left lower left direction, is represented as follows. $$\text{S.}\downarrow =\text{tan}\left(2*{\mathrm{\theta }}_0-90°\right)\text{<figure-callout id="0" label="tan θ" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">tan</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_0/\left(\text{tan}{\mathrm{\theta }}_0-\text{tan}\left(2*{\mathrm{\theta }}_0-90°\right)\right)*\left(\text{x}-\mathrm{\beta }/\text{cos}\left(90°-{\mathrm{\theta }}_0\right)\right)$$

An offset S ↑ upwards of the light beam, which propagates inside the respective dichroic mirror in a left upper direction, is expressed as follows: $$\text{S.}\uparrow =1/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)$$

Here, β indicates the thickness of each dichroic mirror and x indicates the distance between the dichroic mirrors. To compensate for S ↓ and S ↑, S ↓ = S ↑ is desirable, as in 20th shown. Here, θ _{0 is given as θ 0} (BM) in the best mode of execution.

When equations (37) and (38) are applied to β = 1 mm and x = 2.5 mm, the conditions in 20th θ ₀ (BM) = 50 °. That is, the in 20th The configuration shown is the configuration in best mode of execution. 21 according to, however, W is _{maximal at θ 0} = 52 °, which is 2 ° greater than the above θ ₀ (BM). This shows that W can be slightly larger if θ _{0 is} slightly larger than θ ₀ (BM), that is, S ↓ is slightly larger than S ↑ and the light beam spreads gradually in the lower left direction.

As described above, a condition for a significant increase in W from the conventional standard θ ₀ = 45 ° is expressed as follows: $$45°\le {\mathrm{<figure-callout\; id="0"\; label="\theta "\; filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png"\; state="\{\{state\}\}">\theta </figure-callout>}}_0\le 2*{\mathrm{\theta }}_0\left(\text{BM}\right)-45°$$

Taking into account the deviation of 2 °, a more precise condition is expressed as follows: $$45°\le {\mathrm{<figure-callout\; id="0"\; label="\theta "\; filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png"\; state="\{\{state\}\}">\theta </figure-callout>}}_0\le 2*{\mathrm{\theta }}_0\left(\text{BM}\right)-43°$$

[Embodiment 6]

22nd FIG. 13 is a schematic illustration of the light emitting detection device when in the configuration of FIG 6th a size of its light emitting point is relatively large. The diameter of a light emitting point 71 is d = 0.5 mm and the distance p = 1 mm, whereby the size of the light-emitting point is one place larger than in the embodiment 1 . The light emitting point 71 each consists of a cubic reaction cell of 0.5 mm × 0.5 mm × 0.5 mm, the chemiluminescence being generated by an internal chemical reaction. Changes with time in the wavelength of chemiluminescence and its intensity were recorded for each light-emitting point 71 measured to analyze a sample placed in each reaction cell. The focal length of the condenser lens 2 is f = 1 mm, the effective diameter is D = 1 mm, the distance between them is p = 1 mm and the optical distance between the condenser lens 2 and the two-dimensional color sensor 11 is g = 10 mm. As for the arrangement direction of the light emitting points 71 and the condenser lenses 2 As for the light emitting points 71 and the condenser lenses 2 be arranged in a lateral direction, as in FIG 22 (a) shown, but can also be in the same Distances in a direction vertical to the plane of the paper of 22 (a) be arranged. Since no excitation light source is required in the present embodiment, this is omitted in FIG 6th shown long pass filter 10.

When the light emitting point 71 on a sensor surface of the two-dimensional color sensor 11 is mapped as in 22 (a) are set according to equation (1) m = 9 and according to equation (2) d '= 4.5 mm. In this case, the equations (6) and (10) are satisfied, the relative amount of detection light is larger than 400%, and the precise relative amount of detection light is larger than 200%. On the other hand, there is crosstalk between different light emitting points 71 the in 22 (a) light rays 9 shown are significantly large, and none of the equations (16) to (18) are satisfied.

As in 22 (b) is in the center of each condenser lens 2 arranged a pinhole arrangement 73, the respective pinhole 72 having each light emitting point 71 correspond. The diameter d _{0 of} each pinhole 72 is set to d ₀ ≤d, here d ₀ = 0.1 mm. A distance between the respective perforated diaphragms is set to p = 1 mm. As for the arrangement direction of the pinhole diaphragms, the pinhole diaphragms can be arranged in a lateral direction shown in FIG 22 (b) is shown, but can also be at equal intervals in a direction vertical to the paper surface in 22 (a) be arranged. Here, instead of the light emitting point 71 the pinhole 72 considered as a light emitting point. When the pinhole 72 on a sensor surface of the two-dimensional color sensor 11 can be mapped to a pinhole image 74 to yield, as in 22 (b) is the diameter of the pinhole image 74 Equation (2) is set to d '= 0.9 mm accordingly. In this case, equation (18) is satisfied, so that the crosstalk can be set to 0%. In this case, the equations (6) and (10) are satisfied, and the relative amount of detection light is larger than 400%, and the precise relative amount of detection light is larger than 200%. However, the above values relate to a total amount of light that passes through the pinhole 72 goes, and are smaller than those that focus on one from the light-emitting point 71 in 22 (a) refer to the total amount of light emitted.

23 Figure 3 is a schematic representation of another example based on a 22 (b) corresponding detection device is applied. As in 23 (a) is the diameter of a light emitting point 75 d = 0.01 mm, and a distance is p '= 0.1 mm. Further is a focal length of the condenser lens 2 f = 1 mm, an effective diameter D = 1 mm and a distance p = 1. In other words, in contrast to the embodiment described above, the distance between the light-emitting points is different from the distance between the condenser lenses, in this example p '<p. The light emitting points 75 and the condenser lenses 2 can also be in a direction perpendicular to the paper surface 23 (a) be arranged at equal intervals. When the light emitting point 75 on a sensor surface of the two-dimensional color sensor 11 is imaged, the crosstalk continues to increase as light emits from a plurality of light emitting points 75 in a larger area than in 22 (a) is emitted through the respective condenser lenses 2 is bundled.

Hence, as in 23 (b) shown, a pinhole arrangement 73 with respective pinholes 72 between the light emitting point 75 and the corresponding condenser lens 2 arranged. The respective pinhole diaphragms 72 are each with corresponding condenser lenses 2 aligned. The diameter do of each pinhole 72 if d ₀ ≤d, and here d ₀ = 0.1 mm. A distance between the respective pin holes is p = 1 mm. As far as the arrangement direction of the pinhole diaphragms is concerned, the pinhole diaphragms can also be in a direction perpendicular to a plane of the paper 23 (b) be arranged at equal intervals. Here, the light emitting point array and the pinhole array 73 are arranged close enough to each other that each light emitting point is 75 and every pinhole 72 together on the sensor surface of the two-dimensional color sensor 11 can be mapped, creating one image each 76 of the light emitting point and a pinhole image 74 be generated. The diameter of the pinhole image 74 Equation (2) is set to 0.9 mm accordingly, and the crosstalk between the various pinhole images 74 is set to 0% what 22 (b) corresponds to. On the other hand, each condenser lens focuses 2 Light emitted from several (on average two) light emitting points 75 over each pinhole 72 is emitted to on the sensor surface of the two-dimensional color sensor 11 inside each pinhole image 74 the pictures 76 of the light emitting points. Because the diameter of each picture 76 of the light emitting points is set to 0.09 mm and a distance is set to 0.9 mm, occurs between the different images 76 the light-emitting points do not show any crosstalk.

23 (b) Fig. 10 illustrates a configuration in which it is possible to detect light emitted from light emitting points that are spaced closer than the condenser lenses with high sensitivity and little crosstalk in multiple colors. However, this configuration can only recognize a part of the light emitting points among the plurality of light emitting points. In case of 23 (b) only two out of ten light-emitting points are recognized on average. Therefore, all of the light emitting points as shown in 23 (b) can be detected, for example, by successively shifting the relative position of a light-emitting point arrangement and a detection device located behind the pinhole 73 in the direction of the arrow, that is, by scanning the light-emitting points to be detected among a plurality of light-emitting points.

In the above, a multicolor detection device with a two-dimensional color sensor was used. Alternatively, however, a detection device with a two-dimensional monochrome sensor can be used instead of the two-dimensional color sensor. Also, a multicolor detection device with a dichroic arrangement as shown in FIG 7th shown can be used.

24 (a) FIG. 13 is a schematic diagram showing examples of a multicolor detection device having a dichroic arrangement used in the present embodiment, and is a configuration obtained by combining 7 (b) and 23 (a) is obtained. In 24 (a) the laser beams 21, 22, 23 and 24 each have different optical path lengths. If one of the images 25, 26, 27 and 28 of the corresponding light-emitting points on the sensor surface of the two-dimensional sensor 30th is imaged, therefore, no other image of light emitting points is imaged on the sensor surface, resulting in a slight blurring state. There in 7th a light emitting point 75 through a condenser lens 2 is converged and four images 25, 26, 27 and 28 of the light emitting point are obtained, the above blurring condition does not cause crosstalk. On the other hand, since in 24 (a) a variety of light emitting points 75 through a condenser lens 2 are converged and four divided images 25, 26, 27 and 28 of the light emitting point are obtained for each light emitting point, the plurality of images are overlapped 76 of the light-emitting points inside the pinhole image 74 due to the above unsharp condition, which leads to crosstalk.

As in 24 (b) shown, optical path length adjusting members 77, 78 and 79 of different lengths are inserted into the optical paths of the light beams 21, 22 and 23, and the optical path lengths of the laser beams 21, 22, 23 and 24 determined by an optical distance between the condenser lens 2 and the two-dimensional sensor 30th are approximately the same. The optical path length setting element consists of a transparent material whose refractive index is greater than 1. For example, in a material with a refractive index of 2, the optical path length is 2 times greater than in air, even if it is spatially the same. According to the above configuration, it is possible to simultaneously display the images 25, 26, 27 and 28 of the light emitting points on the sensor surface of the two-dimensional sensor 30th image, thereby preventing the blurring state and the crosstalk caused by it.

As described above, the light emitting dot array arranges a detection object at approximately equal intervals, but the detection object may be arranged in a one-dimensional, two-dimensional, or three-dimensional light-emitting distribution. Further, an imaging method can be applied by taking an original light-emitting distribution using a light-emitting detection device as in FIG 23 (b) is reconstructed from the recognition result. 25th illustrates examples of imaging techniques.

As in 25 (a) shown is a light emitting distribution 80 distributed in a two-dimensional form. Here, as an example, there is shown a case where the light emitting distribution 80 represents a character "α". Furthermore, the pinhole arrangement 73 is parallel to the light-emitting point distribution 80 and located close to it. 25 (a) represents in a schematic manner a positional relationship and a size ratio of a plurality of pinhole diaphragms 72 contained in the pinhole assembly 73 and the light emitting point distribution 80 Here are 3 × 3 = 9 pinhole diaphragms 72 arranged two-dimensionally at equal intervals. Since the light-emitting point distribution 80 and every pinhole 72 Are close enough together, light will come from part of the light-emitting point distribution 80 is emitted, which is with a pinhole 72 of 25 (a) overlapped by a light emitting detection device as in FIG 23 (b) through each pinhole 72 recognized.

25 (b) shows 9 pinhole images 74 who have favourited pictures of 9 pinholes 72 are those from the two-dimensional color sensor 11 and 9 partial images 81 of the light-emitting point distribution, the partial images of the through 9 pinhole diaphragms 72 recognized light-emitting point distribution 80 are. As in previous embodiments, each hole 72 shown enlarged, and there is a distance of the hole 72 and a spacing of the hole pattern 74 are the same, the distance between the neighboring hole patterns will be 74 as in 25 (b) shown narrowed, the mutual crosstalk was 0%. Since each sub-image 81 of the light-emitting point distribution in a corresponding pinhole image 74 is accommodated, the mutual crosstalk is also at 0%.

25 (c) represents the relative position of the light-emitting point distribution 80 and pinhole assembly 73 of Fig. 25 (a) after being slid in a lateral direction. Any pinhole 72 compared to Fig. 25 (a) lies over a different part of the light-emitting point distribution 80 . Light emitted from these overlapped parts is as shown in FIG 25 (d) shown mapped and recognized. The image acquisition is repeated by changing the relative position of the light-emitting point distribution 80 and the pinhole array 73 is sequentially (that is, sequentially) shifted in the lateral and longitudinal directions, thereby making it possible to enhance the imaging process of an overall image of the light-emitting point distribution 80 perform. When the relative position of the light-emitting point distribution 80 and the two-dimensional color sensor 11 is set, the overall picture of the light-emitting point distribution 80 can also be mapped without performing image processing.

[Embodiment 7]

One of the objects in implementing the invention is to accurately and easily align each light emitting point with each condenser lens. The present embodiment shows a method for realizing this for a large number of capillaries.

26th Fig. 13 is a cross-sectional diagram schematically showing configuration examples of an apparatus 86 in which a plurality of capillaries 49 , a V-groove arrangement for arranging the plurality of capillaries 49 and the condenser lens assembly 8th are integrated. 26 (a) Figure 10 shows a cross section perpendicular to a longitudinal axis of each capillary 49 at an irradiation position of the laser beam 54; 26 (b) Figure 10 shows a cross section perpendicular to the longitudinal axis of each capillary 49 at a position other than the irradiation position of the laser beam 54; and 26 (c) Figure 11 shows a cross section containing a longitudinal axis of a single capillary. 26 (a) corresponds to a cross section along the line AA of 26 (c) , and 26 (b) corresponds to a cross section along the line BB of 26 (c) .

In the 26th The illustrated device 86 comprises the capillary arrangement with a plurality of capillaries 49 and a dividing device 85. The dividing device 85 is with a V-groove arranging device 84 which is a part including the V-groove arrangement in which a plurality of V-grooves 82 are arranged at a pitch p, and with a condenser lens arrangement device 83 which is a part that makes up the condenser lens assembly 8th contains, in which a variety of condenser lenses 2 are arranged at a distance p. In 26 (a) the central axis of each light emitting point is correct 1 , each V-groove 82 and each condenser lens 2 coincide with each other. The multitude of capillaries 49 can easily be arranged on the same plane at the predetermined distance p by the plurality of capillaries 49 pressed against the V-grooves 82. Further, a structure of the dividing device 85 is set so that each light emitting point 1 showing the irradiation position of each capillary 49 through the laser beam 54, and each condenser lens 2 are at a desired distance from each other. That from the light emitting point 1 emitted light is therefore passed through the condenser lens as desired 2 bundled.

As in 26 (a) is shown in the cross section of the capillary 49 at the light emitting point 1 the condenser lens 2 of the dividing device 85 is present, and the V-groove 82 is not present. As in 26 (b) is shown in the cross section of the capillary 49 on either side of the light emitting point 1 the condenser lens 2 the dividing device 83 does not exist, and the V-groove 82 is present. 26 (c) shows a cross section in the direction of the longitudinal axis of the capillary 49 who have favourited the condenser lens 2 is in the center of divider 85 and V-grooves 82 are on opposite sides of the condenser lens 2 . Depending on the configuration, it is advantageous not only for the high-precision alignment of the capillaries due to the V-groove 82 49 to enable but also to prevent the detection of the light emitting point 1 emitted light is disturbed by the V-groove 82. If a dividing device 85 as described above is prepared in advance, high-precision alignment between the respective light emitting points can be achieved 1 and the respective condenser lenses 2 can be easily achieved by using the multitude of capillaries 49 are pressed against the respective V-grooves 82.

The present embodiment can be combined with the configuration of any of the above embodiments. The dividing device 85, in which the V-groove arrangement device 84 and the condenser lens arrangement device 83 are integrated with each other can be integrally molded by manufacturing processes such as injection molding and embossing, which enables mass production at low cost. In addition can use the V-Groove Placement Device 84 and the condenser lens arrangement device 83 can also be manufactured separately and then connected to one another in order to complete the dividing device 85.

The dividing device is also effective in the absence of a V-groove arrangement. For example, a surface of the divider on the capillary array side may be flat without having a V-groove arrangement. Then, although the arrangement pitches of the plurality of capillaries must be adjusted by other means, it is possible to regulate a distance between each capillary and each condenser lens, that is, a distance between each light emitting point and each condenser lens by keeping each capillary against the above flat Surface of the dividing device is pressed. Alternatively, a structure for adjusting the position of the capillary can be provided in the sub-device, even if the V-groove is not provided.

When the focal length of each condenser lens 2 in a direction parallel to the arrangement direction of the light emitting dot array is defined as f1 and the focal length of which in a vertical direction is defined as f2, f1 and f2 are set as f = fl = f2 in the above embodiments. But it can also be effective to set f1 and f2 as f1 ≠ f2. For example, it is effective when the light emitting point 1 as in the present embodiment in the interior of the capillary 49 is located. Because the capillary 49 has a cylindrical shape, has the capillary 49 in a direction of arrangement of the light-emitting point arrangement on a lens effect, in the direction of its longitudinal axis 49 however, the capillary has no lens effect. To from the light emitting point 1 emitted light with the condenser lens 2 To focus efficiently, it is therefore effective to detect the directional difference in the lens action of the capillary 49 to be canceled, and to this end, it is desirable to set f1 and f2 as f1 ≠ f2, more precisely, fl <f2. This can be achieved simply by removing the surface of the respective condenser lenses 2 a spherical shape is given. Further, the thickness of the lens can be reduced and the fluorescence detection device can be further miniaturized by adding each condenser lens 2 is used as a Fresnel lens. The use of the Fresnel lens is also effective when setting fl = f2.

27 Fig. 10 illustrates a configuration in which between the light emitting dot array and the condenser lens array 8th in 26th the pinhole assembly 73 was added. That is, the pinhole assembly 73 is between the V-groove locator 84 and the condenser lens assembly device 83 and any of the configurations described above is used as the subdevice 86. The pinhole 72 , in the 22 (b) is shown restricts an amount of light from the light emitting point by its presence 71 emitted and from the condenser lens 2 bundled light. In the case of the in 27 illustrated pinhole 72 on the other hand, the diameter do of each pinhole is 72 larger than the diameter d of the light emitting point 1 (d ₀ ≥d) so that the amount of light from the light emitting point 1 emitted and from the condenser lens 2 focused light is not restricted by its presence, as in 27 (a) shown. While the pinhole 72 of 22 (b) effectively serves to determine the diameter of the light emitting point 71 the pinhole is used to reduce this 72 of 27 in addition, the bundling of light other than that from the light-emitting point 1 is emitted to which the condenser lens 2 corresponds to avoid. For example, it is possible to prevent scattered light from the laser beam 54 generated when irradiating the capillary with the laser beam 54 on an outer surface of each capillary 49 arises through the condenser lens 2 is bundled and reaches the sensor, or to reduce it. Or light coming from an adjacent light emitting point 1 can be emitted through the condenser lens 2 be bundled in such a way that it does not reach the sensor or is reduced. This makes it possible from the light emitting point 1 detect emitted light with high sensitivity.

In order to prevent unnecessary light from reaching the sensor, it is also effective to place a colored glass filter between the light emitting point and the sensor. The colored glass filter can be combined with the pinhole, or just one of them can be used. The ray of unnecessary light emitted by the condenser lens 2 is bundled, spreads to the optical axis of the condenser lens 2 (ie the optical axis of the light beam to which that from the light emitting point 1 emitted light through the condenser lens 2 is inclined), which makes it difficult to block the light beam by the long-pass filter or dichroic mirror (because the condenser lens 2 is designed to be from the light emitting point 1 to bundle emitted light into a light beam). In addition, the color glass filter can display an equivalent filter performance even if the angle of incidence of the light is different, whereby the above effect is obtained.

28 represents a different configuration to obtain the same effect as in 27 The condenser lens assembly device 83 , which is part of the dividing device 85, is made of a transparent material such as a glass material and a resin material, as described above. The V-groove Placement device 84 on the other hand consists of an opaque material. The pinholes 87 , which are through holes, are each arranged at a position coincident with the optical axis of each condenser lens 2 of the V-Groove Placement Device 84 cuts so the V-groove locator 84 serves as a pinhole arrangement. According to this configuration, it is possible to manufacture the dividing device 85 more easily than in the case of FIG 27 .

The present invention is not limited to the above-described embodiments, but on the contrary is intended to cover various modifications and equivalent arrangements falling within the spirit and scope of the appended claims. For example, while embodiments have been described in detail in order to describe the present invention in an easily understood manner, it should be understood that the invention is not limited to the disclosed configurations. Further, a part of a configuration of a certain embodiment can be replaced with a configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of an embodiment. Further, with respect to a part of the configuration of each embodiment, it is possible to add, delete, and replace other configurations.

The invention can be further characterized by the following aspects. A first aspect is a light emitting detection device comprising: a condenser lens array arranged with M condenser lenses for respectively converging light emitted from a light emitting point array in which M light emitting points are arranged to form a light beam, where M ≥ 2; at least one sensor on which the M light rays are incident in parallel without being bundled again, where if d is an average value of the effective diameter of the M light-emitting points, f is an average value of the focal length of the M condenser lenses, p is an average value of the intermediate distance of the M condenser lenses p is, and g is an average value of the maximum optical path length between the M condenser lenses and the sensor, d, f, p and g satisfy a certain relationship which is predetermined so that the M light emissions can be detected with little crosstalk or high sensitivity .

According to a further second aspect, the light-emitting detection device can furthermore comprise a pinhole arrangement with M pinholes which are arranged such that they are aligned with the M condenser lenses, the M light-emitting points being formed by light emission of a part of at least one light-emitting region.

According to a third aspect, the light-emitting detection device may further comprise an optical element that deflects the M light beams to generate deflected light beams, or an optical element that refocuses the M light beams each time to generate refocused light beams, the M deflected light beams or the M re-bundled light beams fall parallel to the sensor.

$$\text{f}\ge 1/\left(\text{p}/\text{d}+1\right)*\text{g}\text{,}$$

$$\text{f}\le 2*\text{p}$$

oder $$\text{f}\le \text{p}\text{.}$$

According to fourth to seventh aspects, one of the following relationships can be satisfied for the light emitting detection device: $$\text{f}\ge 1/\left(\left(2*\text{p}\right)/\left(1.27*\text{d}\right)+1\right)*\text{G}\text{,}$$

$$\text{f}\ge 1/\left(\text{p}/\text{d}+1\right)*\text{G}\text{,}$$

$$\text{f}\le 2*\text{p}$$

or $$\text{f}\le \text{p}\text{.}$$

According to an eighth aspect, the light-emitting detection device can also include a dichroic mirror array in which a plurality N of dichroic mirrors in the numerical order 1, 2, ..., N, where N ≥ 2, in a first direction optically between the condenser lens array and the Sensor are arranged, include, wherein normal vectors on front surfaces of the N dichroic mirrors are formed by a sum of a positive component in the first direction and a negative component in a second direction perpendicular to the first direction, the N normal vectors essentially are parallel to each other, the optical axes of the M condenser lenses are substantially parallel to the second direction, and an arrangement direction of the M condenser lenses is substantially parallel to a third direction perpendicular to both the first and second directions.

According to a ninth aspect, in the light emitting detection device, the M light beams can be incident on the dichroic mirror array in parallel along the second direction, a split light beam obtained in the first direction by dividing the M light beams into N different light beams from the dichroic mirror array along the second direction are emitted, and the (MxN) split light beams are incident on the sensor in parallel and are recognized together.

According to a tenth aspect, in the light emitting detection device, when D is an average value of the effective diameter of the M condenser lenses, θ _{0 is} an average value of the angles formed between the N normal vectors and a direction opposite to the second direction, where 0 θ ₀ ≤ 90 °, no is an average value of the refractive index of the base materials of the N dichroic mirrors, α is an average value of the width of the base materials of the N dichroic mirrors, β is an average value of the thickness of the base materials of the N dichroic mirrors, x is an average value of the intermediate distance of the N. dichroic mirror, and yz is an average of the distances by which an end in the second direction of the nth dichroic mirror is displaced in a direction opposite to the second direction opposite to an end in the second direction of the n-1st dichroic mirror , where 2 ≤ n ≤ N, d, f, D, p, g, θ ₀ , N, no, α, β, x, and yz have a certain relationship meet which is predetermined so that the M light emissions can be recognized with the dichroic mirrors.

und $$\left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{w}}/{\text{a}}_{\text{w}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta }+\text{cos}{\mathrm{\theta }}_0/\text{aW}*\text{d'}\le \text{x}\le \left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{L}}/{\text{a}}_{\text{L}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta +}\text{cos}{\mathrm{\theta }}_0/{\text{a}}_{\text{L}}*\text{g}$$

und/oder die folgende Beziehung erfolgt sein: $$0\le \text{yz}\le 2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }\mathrm{.}$$

According to an eleventh and a twelfth aspect, in the light emitting detection device with θ ₁ = sin ^-1 (1 / n ₀ * sinθ ₀ ), θ ₂ = sin ^-1 (1 / n ₀ * sin (90 ° -θ ₀ )) , aw = cosθ ₀ , bw = -cosθ ₀ * tanθ ₁ , a _L = (N-1) * cosθ ₀ + sinθ ₀ , b _L = (N-2) / cosθo * (2 * sin (90 ° -θ ₀ -θ ₂ ) + 1-sin (θ ₀ + θ ₂ )) + (N-2) * sinθ ₀ + 2 * cosθ ₀ , and d '= (gf) / f * d, the following relationships are satisfied: $$\left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{w}}/{\text{a}}_{\text{w}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta }+\text{cos}{\mathrm{\theta }}_0/{\text{a}}_{\text{w}}*\text{D.}\le \text{x}\text{,}$$

and $$\left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{w}}/{\text{a}}_{\text{w}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta }+\text{cos}{\mathrm{\theta }}_0/\text{aW}*\text{d '}\le \text{x}\le \left(\text{sin}{\mathrm{\theta }}_0-{\text{b}}_{\text{L.}}/{\text{a}}_{\text{L.}}*\text{cos}{\mathrm{\theta }}_0\right)*\mathrm{\beta \; +}\text{cos}{\mathrm{\theta }}_0/{\text{a}}_{\text{L.}}*\text{G}$$

and / or the following relationship has occurred: $$0\le \text{Y Z}\le 2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }\mathrm{.}$$

und $$\text{wenn }3\le \text{n}\le \text{N},0\le \text{yz}\le 2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }$$

oder die folgende Beziehung erfüllt sein: $$\text{cos}{\mathrm{\theta }}_0*\mathrm{\alpha }\le \text{x}\le \text{cos}{\mathrm{\theta }}_0*\mathrm{\alpha }+2*\text{sin}{\mathrm{\theta }}_0*\mathrm{\beta }.$$

According to a thirteenth and a fourteenth aspect, the following relationships can also be found $$\text{if n}=2.0\le \text{Y Z}\le 2*\text{cos}{\mathrm{\theta }}_0*\mathrm{\beta },$$

and $$\text{if}3\le \text{n}\le \text{N},0\le \text{Y Z}\le 2*\text{sin}\left(90°-{\mathrm{\theta }}_0-{\mathrm{\theta }}_2\right)/\text{cos}{\mathrm{\theta }}_2*\mathrm{\beta }$$

or the following relationship may be met: $$\text{cos}{\mathrm{\theta }}_0*\mathrm{\alpha }\le \text{x}\le \text{cos}{\mathrm{\theta }}_0*\mathrm{\alpha }+2*\text{<figure-callout id="0" label="sin θ" filenames="DE112016007594B3\_0061.png,DE112016007594B3\_0064.png" state="{{state}}">sin</figure-callout>}{\mathrm{\theta }}_{}{\mathrm{}}_0*\mathrm{\beta }.$$

According to a fifteenth aspect, in the light emitting detection device, an optical path length adjusting member may be provided in an optical path of at least part of the (MxN) split light beams.

According to a sixteenth aspect, in the light-emitting detection device, the M condenser lenses can be arranged two-dimensionally in a plane parallel to a sensor surface of the sensor.

According to a seventeenth aspect, in the light emitting detection device, the sensor may be a color sensor in which plural kinds of pixels are arranged to distinguish different wavelength bands.

According to an eighteenth aspect, the light-emitting detection device can further comprise a mechanism that displaces at least one relative position of the condenser lens arrangement with respect to the M light-emitting points.

According to a nineteenth aspect, the invention can relate to an apparatus in which a V-groove arrangement having a plurality of V-grooves and a condenser lens arrangement having a plurality of condenser lenses are integrated, the V-groove arrangement and the condenser lens arrangement being equally spaced are arranged.

According to a twentieth aspect, in such an apparatus, a pinhole assembly having a plurality of pinholes may be provided between the V-groove assembly and the condenser lens assembly, and the V-groove assembly, the condenser lens assembly and the pinhole assembly may be equally spaced.

List of reference symbols

1

light emitting point

2

Condenser lens

3, 3 '

parallel beam of light

4, 5

Point of light

7th

light-emitting point arrangement

8th

Condenser lens assembly

11

two-dimensional color sensor

17-20

dichroic mirror

30th

two-dimensional sensor

31

Transmission diffraction grating

33

Re-condenser lens

37

two-dimensional sensor

41

wavelength dispersed image

43

prism

45, 48

Image of the light emitting point

49

capillary

53

Laser light source

71

light emitting point

72

Pinhole

74

Pinhole image

75

light emitting point

76

light-emitting point arrangement

77-79

optical path length adjustment element

80

light emitting distribution

83

Condenser lens placement device

84

V-groove placement device

87

Pinhole

Claims (12)

A light emission detection device comprising: a condenser lens array (8) in which M condenser lenses (2) are arranged for respectively converging light emitted from a light emitting point array (7) in which M light emitting points (1) are arranged to generate M light beams to form, where M ≥ 2, an optical element (43) which deflects the M light beams to generate M deflected light beams, and at least one sensor (11) on which the M light beams are incident in parallel, where if d is an average is the effective diameter of the M light emitting points, f is an average value of the focal lengths of the M condenser lenses, p is an average value of the intermediate distances of the M condenser lenses, and g is an average value of the maximum optical path lengths between the M condenser lenses and an end face of the optical element on which the M rays of light are incident, d, f, p and g satisfy the following relationship: $$\text{f}\ge \text{1/}\left(\left(2*\text{p}\right)/\left(1.27*\text{d}\right)+1\right)*\text{G}\text{.}$$

Light emission detection device according to Claim 1 , where the following relationship is met: $$\text{f}\ge 1/\left(\text{p}/\text{d}+1\right)*\text{G}.$$

Light emission detection device according to Claim 1 or 2 , where the following relationship is met: $$\text{f}\le 2*\text{p}$$

Light emission detection device according to one of the Claims 1 to 3 , where the following relationship is met: $$\text{f}\le \text{p}.$$

Light emission detection device according to one of the Claims 1 to 4th , further comprising: a pinhole assembly (73) in which M pinholes (72) are disposed and aligned with the M condenser lenses (2), the M light emitting points (71) being formed by light emission from a portion of at least one light emitting area. A light emission detection device comprising: a condenser lens array (8) in which M condenser lenses (2) are arranged for respectively converging light emitted from a light emitting point array (7) in which M light emitting points (1) are arranged to generate M light beams to form, wherein M ≥ 2, a re-condenser lens arrangement in which M re-condenser lenses (33) are arranged for the respective re-bundling of the M light beams, in order to form re-bundled light beams; at least one sensor (11) on which the M re-bundled light beams are incident in parallel, where, if d is an average value of the effective diameter of the M light-emitting points, f is an average value of the focal lengths of the M condenser lenses, p is an average value of the intermediate distances of the M condenser lenses and g is an average of the maximum optical path lengths between the M condenser lenses and the M Re condenser lenses, d, f, p and g satisfy the following relationship: $$\text{f}\ge \text{1/}\left(\left(2*\text{p}\right)/\left(1.27*\text{d}\right)+1\right)*\text{G}\text{.}$$

Light emission detection device according to Claim 6 , where the following relationship is met: $$\text{f}\ge \text{1/}\left(\text{p / d}+1\right)*\text{G}\text{.}$$

Light emission detection device according to Claim 6 or 7th , where the following relationship is met: $$\text{f}\le 2*\text{p}\text{.}$$

Light emission detection device according to one of the Claims 6 to 8th , where the following relationship is met: $$\text{f}\le \text{p}\text{.}$$

Light emission detection device according to one of the Claims 6 to 9 , further comprising: a pinhole assembly (73) in which M pinholes (72) are disposed and aligned with the M condenser lenses (2), the M light emitting points (71) being formed by light emission from a portion of at least one light emitting area. Light emission detection device according to one of the Claims 1 to 10 , further comprising: a capillary arrangement with a plurality of capillaries (49) aligned on a plane and a device (85) for holding the capillary arrangement, the light-emitting point arrangement (7) being formed from individual light-emitting points from the capillary arrangement (49) and the condenser lens arrangement ( 8) is integrated into the device (85). Light emission detection device according to Claim 11 , further comprising: a pinhole arrangement (73) which is arranged between the capillary arrangement and the condenser lens arrangement (8) and in which M pinholes (72) are provided which are aligned with the M condenser lenses (2).

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