Fluorescence reader and fluorescence reading method

Abstract

SOLUTION: Provided is a fluorescence reader for acquiring the fluorescence of a micro-array. The fluorescence reader comprises a light receiving optical system for reflecting the fluorescence radiated from the micro-array a number of times by a plurality of reflectors and forming an image on an image forming plane, and an optical sensor located on the image forming plane. The fluorescence reader may be of a telecentric optical system where the imaging plane and the image forming plane of the micro-array are of the same scale. The plurality of reflectors may include a convex mirror and a concave mirror. The plurality of reflectors may only include one convex mirror and one concave mirror.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a fluorescence reading device and a fluorescence reading method. [Background technology]

[0002] Patent Document 1 states, "Fluorescence from biochip 60 generated by excitation with laser light is incident on CCD camera 13 via dichroic mirror 3, dichroic mirror 23, barrier filter 11, and optical system 12." (Paragraph 0040) Patent Document 2 states, "Relay optical system 51 has first mirror 46 having positive power, second mirror 47 having negative power, and third mirror 48 having positive power. Light emitted from rod integrator 45 is totally reflected by TIR prism 53 via first mirror 46, second mirror 47, and third mirror 48 of relay optical system 51, and illuminates DMD (first optical modulation element) 49, which is an image display panel." (Paragraph 0042) [Prior art document] [Patent documents] [Patent Document 1] JP 2007-093250 [Patent Document 2] JP 2020-052342 Summary of the Invention

[0003] A first aspect of the present invention provides a fluorescence reading device for acquiring fluorescence from a microarray, comprising: a light receiving optical system that reflects the fluorescence emitted from the microarray multiple times using multiple reflectors to form an image on an imaging plane; and an optical sensor positioned on the imaging plane.

[0004] The fluorescence reading device may be a telecentric optical system in which the imaging plane of the microarray and the image forming plane have the same magnification.

[0005] In any of the above fluorescence reading devices, the light receiving optical system may not include a lens.

[0006] In any of the above fluorescence reading devices, the plurality of reflectors may include a convex mirror and a concave mirror.

[0007] In any of the above fluorescence reading devices, the convex mirror and the concave mirror may be spherical mirrors.

[0008] In any of the above fluorescence reading devices, the plurality of reflectors may include only one of the convex mirrors and one of the concave mirrors.

[0009] In any of the above fluorescence reading devices, the plurality of reflectors may include only two of the convex mirrors and two of the concave mirrors.

[0010] In any of the above fluorescence reading devices, the plurality of reflectors may include only one of the convex mirrors, one of the concave mirrors, and one retroreflector.

[0011] In any of the above fluorescence reading devices, the plurality of reflectors may include only one of the convex mirrors, one of the concave mirrors, and one right-angle reflector.

[0012] A second aspect of the present invention provides a fluorescence reading method for acquiring fluorescence from a microarray, the fluorescence reading method comprising: forming an image of the fluorescence emitted from the microarray on an imaging plane by reflecting the fluorescence multiple times by multiple reflectors in a light receiving optical system; and receiving the fluorescence with an optical sensor positioned on the imaging plane.

[0013] The above summary of the invention does not list all of the features of the present invention, and subcombinations of these features may also be inventions. [Brief explanation of the drawings]

[0014] [Figure 1] 1 is a schematic diagram showing an outline of a fluorescence reading device 100 according to a first embodiment. [Figure 2]1 is a schematic diagram for explaining the configuration of a light receiving optical system 300 in a fluorescence reading device 100 according to a first embodiment. [Figure 3] The results of a simulation using the fluorescence reading device 100 according to the first embodiment are shown. [Figure 4] FIG. 10 is a schematic diagram for explaining the configuration of a light receiving optical system 400 in a fluorescence reader 102 according to a second embodiment. [Figure 5] The results of a simulation using the fluorescence reader 102 according to the second embodiment are shown. [Figure 6] The results of a simulation using the fluorescence reader 102 according to the second embodiment are shown. [Figure 7] FIG. 10 is a schematic diagram for explaining the configuration of a light receiving optical system 500 in a fluorescence reader 103 according to a third embodiment. [Figure 8] FIG. 10 is a schematic diagram for explaining the configuration of a light receiving optical system 500 in a fluorescence reader 103 according to a third embodiment. [Figure 9] The results of a simulation using the fluorescence reader 103 according to the third embodiment are shown. [Figure 10] The results of a simulation using the fluorescence reader 103 according to the third embodiment are shown. [Figure 11] FIG. 10 is a schematic diagram for explaining the configuration of a light receiving optical system 600 in a fluorescence reader 104 according to a fourth embodiment. [Figure 12] FIG. 10 is a schematic diagram for explaining the configuration of a light receiving optical system 600 in a fluorescence reader 104 according to a fourth embodiment. [Figure 13] 10 shows the results of a simulation using the fluorescence reader 104 according to the fourth embodiment. [Figure 14] 10 shows the results of a simulation using the fluorescence reader 104 according to the fourth embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present invention will be described below through embodiments of the invention, but the following embodiments do not limit the scope of the invention according to the claims. Furthermore, not all of the combinations of features described in the embodiments are necessarily essential to the solution of the invention.

[0016] FIG. 1 is a schematic diagram showing an outline of a fluorescence reading device 100 according to a first embodiment. FIG. 1 shows an X-axis, a Y-axis, and a Z-axis that are orthogonal to each other. The X-axis extends left and right into the paper, the Y-axis extends deep into the paper, and the Z-axis extends up and down into the paper. The corresponding X, Y, and Z-axes are also shown in the subsequent figures, and redundant explanations will be omitted.

[0017] The fluorescence reading device 100 acquires fluorescence from the microarray 1. The fluorescence reading device 100 is a telecentric optical system in which the imaging plane of the microarray 1 and the image forming plane are the same magnification. The fluorescence reading device 100 includes a light receiving optical system 300 and a CCD camera 107. The CCD camera 107 is an example of an optical sensor, and is located on the image forming plane of the light receiving optical system 300. Note that an optical sensor other than a CCD camera may be located on the image forming plane of the light receiving optical system 300.

[0018] The fluorescence reader 100 according to the first embodiment further includes, for example, a laser light source 101, an irradiation optical system 102, a mirror 103, a dichroic mirror 104, a stage 105, a first barrier filter 201, and a second barrier filter 208.

[0019] In the fluorescence reading method according to the first embodiment, as shown by the arrows in Fig. 1, laser light emitted from a laser light source 101 is expanded and shaped into a beam by an irradiation optical system 102 so as to uniformly irradiate the observation surface of a microarray 1, which is a sample. In this embodiment, the microarray 1 is, for example, a DNA microarray. In addition to a DNA microarray, the microarray 1 may also be, for example, a protein microarray or a sugar chain microarray.

[0020] The light beam shaped by the irradiation optical system 102 is reflected by a mirror 103, and then by a dichroic mirror 104, to become irradiation light that irradiates the microarray 1 placed on a stage 105. Fluorescent molecules to be observed by the fluorescence reading device 100 are arranged on the light receiving surface of the microarray 1.

[0021] As shown by the arrows in Figure 1, where the chief ray 210 is shown, the fluorescent molecules on the microarray 1 are excited by the irradiated light and emit fluorescence. The fluorescence from the fluorescent molecules has a very wide divergence angle and is emitted in all directions. The fluorescence from the fluorescent molecules on the microarray 1 passes through the dichroic mirror 104, is imaged on the sensor surface of the CCD camera 107 by the light-receiving optical system 300, and is received by the CCD camera 107, i.e., is detected as an image by the CCD camera 107. The dichroic mirror 104 transmits light in the wavelength range of the fluorescence from the fluorescent molecules and reflects other wavelength ranges, such as the wavelength range of the illumination light. The sensor surface of the CCD camera 107 coincides with the image-forming surface mentioned above.

[0022] Before entering the light-receiving optical system 300, the fluorescence from the fluorescent molecules on the microarray 1 passes through a first barrier filter 201 arranged between the light-receiving optical system 300 and the dichroic mirror 104. The light from the microarray 1 includes not only fluorescence emitted from fluorescent molecules excited by light irradiating the microarray 1, but also light such as reflected light from the irradiating light and autofluorescence generated by substances in the sample other than the target fluorescent molecules and the substrate of the microarray. The first barrier filter 201 transmits only light in the fluorescence wavelength band of the fluorescent molecules on the microarray 1 and blocks light other than the fluorescence of the fluorescent molecules.

[0023] Before being imaged on the sensor surface of the CCD camera 107, the fluorescence emitted from the light-receiving optical system 300 passes through a second barrier filter 208 disposed between the light-receiving optical system 300 and the CCD camera 107. Similar to the first barrier filter 201, the second barrier filter 208 transmits only light in the fluorescence wavelength band of the fluorescent molecules on the microarray 1 and blocks light other than the fluorescence of the fluorescent molecules.

[0024] In this way, the dichroic mirror 104, the first barrier filter 201 and the second barrier filter 208 are installed to extract only the fluorescent wavelengths of the fluorescent molecules to be observed in the microarray 1, and the transmission wavelength bands are designed according to the fluorescent molecules.

[0025] The light receiving optical system 300 reflects the fluorescence emitted from the microarray 1 multiple times using multiple reflectors to form an image on an imaging plane. The light receiving optical system 300 does not include a lens. The multiple reflectors include a convex mirror 301 and a concave mirror 302. In this embodiment, the convex mirror 301 and the concave mirror 302 are spherical mirrors. In this embodiment, the multiple reflectors in the light receiving optical system 300 include only one convex mirror 301 and one concave mirror 302.

[0026] An optical system consisting of spherical convex mirror 301 and concave mirror 302 may also be referred to as an Offner optical system, an Offner reflector optical system, or the like. While convex mirror 301 and concave mirror 302 are spherical mirrors, the multiple reflectors in light-receiving optical system 300 may include reflectors other than spherical mirrors, as long as they can reflect the fluorescence emitted from microarray 1 multiple times and form an image on the image plane. For example, the multiple reflectors in light-receiving optical system 300 may include retroreflectors or right-angle reflectors, which will be described in multiple embodiments below. For example, the multiple reflectors in light-receiving optical system 300 may include a spherical mirror in which a portion of the reflection area, e.g., a reflection area that has little effect on detecting an image of fluorescence from fluorescent molecules with CCD camera 107, is aspherical, i.e., the reflector is essentially a spherical mirror. A retroreflector may also be referred to as a retroreflector.

[0027] Figure 2 is a schematic diagram for explaining the configuration of the light receiving optical system 300 in the fluorescence reading device 100 of the first embodiment. Figure 2 shows the light beam of fluorescence emitted from fluorescent molecules on the microarray 1, with the direction of light travel indicated by an arrow on the chief ray. The same applies to the subsequent figures, and redundant explanations will be omitted.

[0028] The observation surface of the microarray 1 is located at the light-receiving surface focal position 310 of the light-receiving optical system 300. In the light-receiving optical system 300, the fluorescence emitted from the microarray 1 is first reflected by the concave mirror 302, then by the convex mirror 301, and again by the concave mirror 302, and is detected by the CCD camera 107 located at the image-forming surface focal position 320. As described above, the fluorescence reading device 100 is a telecentric optical system in which the imaging surface of the microarray 1 and the image-forming surface are the same magnification, and the image is neither magnified nor reduced.

[0029] 1 and 2, the fluorescence reading device 100 according to the first embodiment comprises a plurality of reflectors, i.e., a plurality of mirror components, that are not affected by chromatic aberration. This allows the fluorescence reading device 100 to be applied to, for example, multi-wavelength microarray detection, and to obtain with high precision an image of a microarray 1 on which fluorescent molecules are arranged.

[0030] Fig. 3 shows the results of a simulation using the fluorescence reader 100 according to the first embodiment. More specifically, Fig. 3 shows the results of a simulation of the configuration of the light receiving optical system 300 in the fluorescence reader 100 according to the first embodiment, where the radius of the spherical mirror is R, the radius of the concave mirror 302 is R=150, the radius of the convex mirror 301 is R=75, and the numerical aperture is NA=0.22.

[0031] Each graph in Figure 3 shows the phase shift [λ] of fluorescence detected at multiple coordinate positions on the sensor surface of CCD camera 107, with the center of the sensor surface as the origin. More specifically, the graph shows the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions at three points on the sensor surface shifted from the origin by [X, Y] = [0.00, 1.00], [0.00, 0.00], and [0.00, -1.00], i.e., when the angle of view is widened. The width of the angle of view in each graph corresponds to the size of the aperture of the optical system.

[0032] According to the simulation results in Fig. 3, in the first embodiment, the positive and negative phase shifts are reversed between the tangential and sagittal directions, i.e., the aberration is dominated by astigmatism and appears centrosymmetrically. Note that, because the phase shift is centrosymmetric, the graph for the negative angle of view in the sagittal direction is not shown.

[0033] For comparison with the fluorescence reader 100 of this embodiment, a comparative example of a fluorescence reader is assumed, which includes a relay lens optical system with two triplet lenses each consisting of three lenses, instead of the light-receiving optical system 300. More specifically, the relay lens optical system of the comparative fluorescence reader includes a triplet lens consisting of three lenses as a light-receiving lens group and a triplet lens consisting of three lenses as an imaging lens group, in order to image the spatial distribution of fluorescence from fluorescent molecules on the microarray onto the sensor surface of the CCD camera to produce a fluorescence image. The triplet lens has the function of correcting monochromatic aberrations such as spherical aberration, coma aberration, and astigmatism, as well as chromatic aberration, which is affected by the wavelength dependence of the refractive index, and corrects deviations from ideal imaging to generate an image with minimal blur.

[0034] As mentioned above, the fluorescence of fluorescent molecules on the microarray is emitted in all directions. Therefore, in observations using very weak light, such as fluorescence observations, the fluorescence reading device of the comparative example enables highly sensitive measurement by receiving the fluorescence emitted in all directions over as wide an area as possible with the light-receiving lens group of the relay lens optical system. According to the fluorescence reading device of the comparative example, the light-receiving range of the fluorescence emitted in all directions is determined by the numerical aperture NA of the first lens in the light-receiving lens group, onto which the fluorescence first enters, and NA is expressed as NA = n·sinθ. Here, n is the refractive index of the medium between the microarray and the first lens, and θ is the maximum angle of light incident from the microarray on the first lens. The medium in question is air, so n = 1.

[0035] The fluorescence reading device of this comparative example requires a relay lens optical system consisting of six lenses to correct aberrations, resulting in high costs. Furthermore, to detect fluorescence with high sensitivity, a relay lens optical system with a high NA receiving lens group is required. To increase the NA, it is necessary to shorten the focal length between the microarray and the first lens or use a first lens with a large effective diameter. However, the relay lens optical system requires the installation of a dichroic mirror and a first filter between the microarray and the first lens. This focal length cannot be narrowed because it depends on the component size and the space occupied by the installation jig. On the other hand, if a first lens with a large effective diameter is used, the lens diameter must be custom-made, resulting in high component costs. In this case, the other five lenses in the relay lens optical system, which depend on the first lens diameter, must also be custom-made, resulting in high component costs.

[0036] In the comparative fluorescence reader, chromatic aberration also occurs because the refractive index of the lens medium is a function of wavelength. In the comparative fluorescence reader, there is a theoretical limit to the correction of chromatic aberration, and in devices that detect a wide wavelength range or observe multicolor fluorescence, the correction of chromatic aberration is insufficient, resulting in blurred images.

[0037] The fluorescence reading device of the comparative example also has a relay lens optical system with six components, which requires a high level of expertise during device assembly and significantly increases the time required for optical axis adjustment. Furthermore, the optical axis adjustment requires a significant amount of labor, both during regular device maintenance and during irregular maintenance to correct optical axis abnormalities. Furthermore, as the number of components increases, the risk of optical axis misalignment, which depends on component misalignment, increases. Furthermore, relay lens optical systems using lenses require optical design that takes wavelength into account as a parameter, making the design difficult. Furthermore, in relay lens optical systems using lenses, ghosting can occur due to reflections on the lens surfaces, and the configuration of the relay lens optical system increases the size of the device in the optical axis direction.

[0038] In contrast, the fluorescence reading device 100 of this embodiment includes the above-mentioned light-receiving optical system 300, unlike the relay lens optical system of the fluorescence reading device of the comparative example. According to the fluorescence reading device 100 of this embodiment, for example, the number of parts in the light-receiving optical system 300 can be reduced to two, a convex mirror 301 and a concave mirror 302, thereby reducing the number of parts compared to the fluorescence reading device of the comparative example. This simplifies the optical axis adjustment, which requires a high level of expertise and time during device assembly and maintenance, and also reduces the risk of optical axis misalignment.

[0039] Furthermore, in the fluorescence reading device 100 of this embodiment, the light receiving optical system 300 is wavelength independent, unlike the relay lens optical system of the comparative example, and therefore optical design is easy. Furthermore, in the fluorescence reading device 100 of this embodiment, the light receiving optical system 300 does not include a lens, and therefore there is no influence of ghosting due to the lens. Furthermore, in the fluorescence reading device 100 of this embodiment, the device size in the optical axis direction of the light receiving optical system 300 can be made smaller than that of the relay lens optical system in the fluorescence reading device of the comparative example.

[0040] Furthermore, the fluorescence reading device 100 according to this embodiment does not require custom lenses as in the fluorescence reading device of the comparative example in order to detect fluorescence with high sensitivity, and instead uses mirror components that are less expensive than custom lenses, thereby reducing costs.

[0041] 3 shows that the amount of aberration in the tangential direction within a 3 mm square field of view is approximately 2λ, while the amount of aberration in the fluorescence reader of the comparative example is approximately 1λ, so the amount of aberration in the fluorescence reader 100 of this embodiment is twice that of the comparative example. Within this acceptable range of aberration, the fluorescence reader 100 of this embodiment is able to eliminate the effects of chromatic aberration and reduce the number of parts in the light-receiving optical system 300 to two, the convex mirror 301 and the concave mirror 302, thereby achieving cost savings compared to the fluorescence reader of the comparative example, which uses six lenses.

[0042] 4 is a schematic diagram illustrating the configuration of a light receiving optical system 400 in a fluorescence reader 102 according to the second embodiment. The fluorescence reader 102 according to the second embodiment differs from the fluorescence reader 100 according to the first embodiment in that it includes a light receiving optical system 400 instead of the light receiving optical system 300. Other components of the fluorescence reader 102 according to the second embodiment are similar to those of the fluorescence reader 100 according to the first embodiment, and therefore the same reference numerals as those of the corresponding components in the fluorescence reader 100 according to the first embodiment are used, and redundant explanations will be omitted.

[0043] Similar to the light receiving optical system 300, the light receiving optical system 400 reflects the fluorescence emitted from the microarray 1 multiple times using multiple reflectors to form an image on an imaging plane, but does not include a lens. The multiple reflectors include a convex mirror and a concave mirror. In this embodiment, the multiple reflectors in the light receiving optical system 400 include only two convex mirrors 301-1 and 301-2 and two concave mirrors 302-1 and 302-2.

[0044] 4, in light receiving optical system 400, a set of one convex mirror 301-1 and one concave mirror 302-1 and a set of one convex mirror 301-2 and one concave mirror 302-2 are arranged at conjugate positions. Each of these sets has the same configuration as light receiving optical system 300 in the first embodiment.

[0045] The observation surface of the microarray 1 is located at the light-receiving surface focal position 310 of the light-receiving optical system 400. In the light-receiving optical system 400, the fluorescence emitted from the microarray 1 is first reflected by the concave mirror 302-1, then by the convex mirror 301-1, and again by the concave mirror 302-1 to form an image at the focal position. Thereafter, it is reflected by the concave mirror 302-2, then by the convex mirror 301-2, and again by the concave mirror 302-2 to be detected by the CCD camera 107 located at the image-forming surface focal position 320.

[0046] 5 and 6 show the results of a simulation using the fluorescence reader 102 according to the second embodiment. More specifically, the results of a simulation of the configuration of the light receiving optical system 400 in the fluorescence reader 102 according to the second embodiment, where the radius of the spherical mirror is R, the radii of the concave mirrors 302-1 and 302-2 are R=150, the radii of the convex mirrors 301-1 and 301-2 are R=75, and the numerical aperture is NA=0.22, are shown in FIGS.

[0047] 5 and 6 show the phase shift [λ] of fluorescence detected at multiple coordinate positions on the sensor surface of CCD camera 107, with multiple points on the sensor surface as origins. More specifically, each graph in Fig. 5 shows the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions, i.e., when the angle of view is widened, at three points on the sensor surface that are respectively shifted from the origins at [X, Y] = [0, 0], [0, -1.5], and [0, 1.5], by [X, Y] = [0.00, 1.00], [0.00, 0.00], respectively. Each graph in Figure 6 shows the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions, i.e., when the angle of view is widened, at three points on the sensor surface, [X, Y] = [0, 0], [-2, -1.5], and [2, 1.5], respectively, from the origin. The graphs in Figure 6 show the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions, i.e., when the angle of view is widened. The width of the angle of view in each graph corresponds to the aperture size of the optical system. Note that, as in Figure 3, the phase shift is centrosymmetric in Figure 5, and therefore the graphs for the negative angle of view in the sagittal direction are omitted.

[0048] The simulation results shown in FIGS. 5 and 6 indicate that in the second embodiment, aberration is corrected across the entire field of view, with the maximum aberration within a 3 mm square field of view being approximately 1λ. Therefore, the fluorescence reader 102 according to the second embodiment can achieve aberrations equivalent to those of the comparative fluorescence reader described above. The fluorescence reader 102 according to this embodiment has the same advantages as the fluorescence reader 100 according to the first embodiment. The fluorescence reader 102 according to this embodiment, for example, can eliminate the effects of chromatic aberration and reduce the number of components in the light-receiving optical system 400 to four: two convex mirrors 301-1 and 301-2 and two concave mirrors 302-1 and 302-2. This reduces costs compared to the comparative fluorescence reader using six lenses. The fluorescence reader 102 according to this embodiment can also further reduce monochromatic aberrations compared to the fluorescence reader 100 according to the first embodiment.

[0049] Fig. 7 is a schematic diagram illustrating the configuration of the light receiving optical system 500 in the fluorescence reading device 103 of the third embodiment. Fig. 8 is a schematic diagram illustrating the configuration of the light receiving optical system 500 in the fluorescence reading device 103 of the third embodiment. The fluorescence reading device 103 of the third embodiment differs from the fluorescence reading device 100 of the first embodiment in that it includes a light receiving optical system 500 instead of the light receiving optical system 300. Other configurations of the fluorescence reading device 103 of the third embodiment are similar to those of the fluorescence reading device 100 of the first embodiment, and therefore the same reference numbers as those of the corresponding configurations in the fluorescence reading device 100 of the first embodiment are used, and redundant explanations will be omitted.

[0050] Similar to the light receiving optical system 300, the light receiving optical system 500 reflects the fluorescence emitted from the microarray 1 multiple times using multiple reflectors to form an image on an imaging plane, but does not include a lens. The multiple reflectors include a convex mirror and a concave mirror. In this embodiment, the multiple reflectors in the light receiving optical system 500 include only one convex mirror 301, one concave mirror 302, and one retroreflector 303.

[0051] 7 and 8, one convex mirror 301 and one concave mirror 302 in the light-receiving optical system 500 have the same configuration as the light-receiving optical system 300 in the first embodiment. Unlike the light-receiving optical system 300, the light-receiving optical system 500 causes the fluorescence from the fluorescent molecules to pass through the convex mirror 301 and the concave mirror 302 twice, once as an optical path on the light-receiving side and once as an optical path on the imaging side.

[0052] The observation surface of the microarray 1 is located at the light-receiving surface focal position 310 of the light-receiving optical system 500. In the light-receiving optical system 500, the fluorescence emitted from the microarray 1 is first reflected by the concave mirror 302, then by the convex mirror 301, and again by the concave mirror 302, before entering the retroreflector 303. The fluorescence reflected by the retroreflector 303 is reflected by the concave mirror 302, then by the convex mirror 301, and again by the concave mirror 302, and is detected by the CCD camera 107 located at the image-forming surface focal position 320.

[0053] The retroreflector 303 may be composed of one mirror component, or may be composed of, for example, three mirror components. In this embodiment, the retroreflector 303 is a corner cube mirror composed of one mirror component with three flat surfaces. In FIG. 8, the retroreflector 303 is shown divided into three parts simply for clarity of explanation.

[0054] 9 and 10 show the results of a simulation using the fluorescence reader 103 according to the third embodiment. More specifically, the results of a simulation of the configuration of the light receiving optical system 500 in the fluorescence reader 103 according to the third embodiment, where the radius of the spherical mirror is R, the radius of the concave mirror 302 is R=150, the radius of the convex mirror 301 is R=75, and the numerical aperture is NA=0.22, are shown in FIGS.

[0055] 9 and 10 show the phase shift [λ] of fluorescence detected at multiple coordinate positions on the sensor surface of CCD camera 107, with multiple points on the sensor surface as origins. More specifically, each graph in Fig. 9 shows the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions at three points on the sensor surface that are shifted from the origins, that is, by [X, Y] = [0, 0], [0, -1.5], and [0, 1.5], respectively, by [X, Y] = [0.00, 1.00], [0.00, -1.00], and [0.00, 0.00], i.e., when the angle of view is widened. The graphs in Figure 10 show the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions at three points on the sensor surface, [X, Y] = [1.00, 1.00], [-1.00, -1.00], and [0.00, 0.00], respectively, from the origin at [X, Y] = [0, 0], [-2, -1.5], and [2, 1.5]. In other words, the angle of view is widened. The width of the angle of view in each graph corresponds to the aperture size of the optical system.

[0056] According to the simulation results shown in FIG. 10 , in the third embodiment, as in the first embodiment, the phase shifts in the tangential and sagittal directions are reversed, i.e., astigmatism is dominant and the aberration is centrally symmetric. The simulation results shown in FIGS. 9 and 10 indicate that the amount of aberration within a 3 mm square field of view is approximately 2λ in the tangential direction and approximately 1λ in the sagittal direction. The amount of tangential aberration in the fluorescence reader 103 according to this embodiment is twice that of the comparative example. The fluorescence reader 103 according to this embodiment has the same effects as the fluorescence reader 100 according to the first embodiment. The fluorescence reader 103 according to this embodiment can, within an acceptable range of aberration, eliminate the effects of chromatic aberration, and reduce the number of components in the light receiving optical system 500 to three: the convex mirror 301, the concave mirror 302, and the retroreflector 303. This reduces costs compared to the fluorescence reader of the comparative example, which uses six lenses. Furthermore, the fluorescence reading device 103 according to this embodiment can further reduce monochromatic aberration compared to the fluorescence reading device 100 according to the first embodiment.

[0057] Fig. 11 is a schematic diagram illustrating the configuration of a light receiving optical system 600 in a fluorescence reader 104 of the fourth embodiment. Fig. 12 is a schematic diagram illustrating the configuration of the light receiving optical system 600 in the fluorescence reader 104 of the fourth embodiment. The fluorescence reader 104 of the fourth embodiment differs from the fluorescence reader 100 of the first embodiment in that it includes a light receiving optical system 600 instead of the light receiving optical system 300. Other configurations of the fluorescence reader 104 of the fourth embodiment are similar to those of the fluorescence reader 100 of the first embodiment, and therefore the same reference numbers as those of the corresponding configurations in the fluorescence reader 100 of the first embodiment are used, and redundant explanations will be omitted.

[0058] The light receiving optical system 600, like the light receiving optical system 300, reflects the fluorescence emitted from the microarray 1 multiple times using multiple reflectors to form an image on an imaging plane, but does not include a lens. The multiple reflectors include a convex mirror and a concave mirror. In this embodiment, the multiple reflectors in the light receiving optical system 600 include only one convex mirror 301, one concave mirror 302, and one right-angle reflector 304.

[0059] 11 and 12, one convex mirror 301 and one concave mirror 302 in the light-receiving optical system 600 have the same configuration as the light-receiving optical system 300 in the first embodiment. Unlike the light-receiving optical system 300, the light-receiving optical system 600 causes the fluorescence from the fluorescent molecules to pass through the convex mirror 301 and the concave mirror 302 twice, once as an optical path on the light-receiving side and once as an optical path on the imaging side.

[0060] The observation surface of the microarray 1 is located at the light-receiving surface focal position 310 of the light-receiving optical system 600. In the light-receiving optical system 600, the fluorescence emitted from the microarray 1 is first reflected by the concave mirror 302, then by the convex mirror 301, and again by the concave mirror 302, before entering the right-angle reflector 304. The fluorescence reflected by the right-angle reflector 304 is reflected by the concave mirror 302, then by the convex mirror 301, and again by the concave mirror 302, and is detected by the CCD camera 107 located at the image-forming surface focal position 320.

[0061] Note that right-angle reflector 304 may be composed of one mirror component, or may be composed of, for example, two mirror components. The angle of incidence of the fluorescence onto the mirror component of right-angle reflector 304 is not particularly limited as long as it can reflect the fluorescence toward concave mirror 302. In this embodiment, right-angle reflector 304 is composed of two mirror components, each consisting of two plane mirrors arranged at an incident angle of 90 degrees.

[0062] 13 and 14 show the results of a simulation using the fluorescence reader 104 according to the fourth embodiment. More specifically, the results of a simulation of the configuration of the light receiving optical system 600 in the fluorescence reader 104 according to the fourth embodiment, where the radius of the spherical mirror is R, the radius of the concave mirror 302 is R=150, the radius of the convex mirror 301 is R=75, and the numerical aperture is NA=0.22, are shown in FIGS.

[0063] 13 and 14 show the phase shift [λ] of fluorescence detected at multiple coordinate positions on the sensor surface of CCD camera 107, with multiple points on the sensor surface as origins. More specifically, each graph in Fig. 13 shows the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions at three points on the sensor surface that are shifted from the origins, that is, by [X, Y] = [0, 1.5], [0, 0], and [0, -1.5], respectively, by [X, Y] = [0.00, 1.00], [0.00, 0.00], and [0.00, -1.00], i.e., when the angle of view is widened. The graphs in Figure 14 show the change in phase shift [λ] when the coordinates are further shifted in the tangential and sagittal directions at three points on the sensor surface, [X, Y] = [-2, -1.5], [0, 0], and [2, 1.5], respectively, from the origin on the sensor surface by [X, Y] = [1.00, 1.00], [0.00, 0.00], and [-1.00, -1.00], i.e., when the angle of view is widened. The width of the angle of view in each graph corresponds to the aperture size of the optical system.

[0064] The simulation results shown in FIG. 13 indicate that in the fourth embodiment, aberration is corrected across the entire field of view, with the maximum aberration within a 3 mm square field of view being approximately 1λ. Therefore, the fluorescence reader 104 according to the fourth embodiment can achieve aberrations equivalent to those of the comparative fluorescence reader described above. The fluorescence reader 104 according to this embodiment has the same advantages as the fluorescence reader 100 according to the first embodiment. The fluorescence reader 104 according to this embodiment, for example, can eliminate the effects of chromatic aberration and reduce the number of components in the light receiving optical system 600 to four: the convex mirror 301, the concave mirror 302, and the right-angle reflector 304 consisting of two plane mirrors. This reduces costs compared to the comparative fluorescence reader using six lenses. The fluorescence reader 103 according to this embodiment can also further reduce monochromatic aberrations compared to the fluorescence reader 100 according to the first embodiment. If the right-angle reflector 304 is configured from one component, the number of components in the light receiving optical system 600 can be reduced to three.

[0065] Although the present invention has been described above using embodiments, the technical scope of the present invention is not limited to the scope described in the above embodiments. It will be apparent to those skilled in the art that various modifications and improvements can be made to the above embodiments. It is clear from the claims that such modifications and improvements can also be included within the technical scope of the present invention.

[0066] It should be noted that the execution order of each process, such as operations, procedures, steps, and stages, in the devices, systems, programs, and methods shown in the claims, specifications, and drawings is not specifically stated as "before," "prior to," etc., and that the processes can be performed in any order unless the output of a previous process is used in a subsequent process. Even if the operational flow in the claims, specifications, and drawings is described using "first," "next," etc. for convenience, this does not mean that the processes must be performed in this order. [Explanation of symbols]

[0067] 1. Microarray 100 Fluorescence reader 101 Laser light source 102 Irradiation optical system 103 Mirror 104 Dichroic Mirror 105 Stages 107 CCD camera 201 First barrier filter 208 Second Barrier Filter 210 Chief ray 300 Light receiving optical system 301 Convex mirror 302 concave mirror 310 Photosensitive surface focal position 320 Image plane focal position 102 Fluorescence reader 400 Light receiving optical system 301-1 Convex mirror 302-1 Concave mirror 301-2 Convex mirror 302-2 Concave mirror 103 Fluorescence Reader 500 Receiving optical system 303 Retroreflector 104 Fluorescence Reader 600 Light receiving optical system 304 Right Angle Reflector

Claims

1. A fluorescence reader for acquiring fluorescence from a microarray, comprising: a light receiving optical system that reflects the fluorescence emitted from the microarray multiple times using multiple reflectors to form an image on an image forming surface; an optical sensor located on the imaging plane; A fluorescence reading device comprising:

2. The imaging surface of the microarray and the image forming surface are a telecentric optical system with a unit magnification.

2. The fluorescence reader of claim 1.

3. The light receiving optical system does not include a lens.

2. The fluorescence reader of claim 1.

4. the plurality of reflectors include a convex mirror and a concave mirror; 4. A fluorescence reading device according to claim 1.

5. the convex mirror and the concave mirror are spherical mirrors; 5. The fluorescence reader of claim 4.

6. the plurality of reflectors include only one convex mirror and one concave mirror; 5. The fluorescence reader of claim 4.

7. the plurality of reflectors include only two of the convex mirrors and two of the concave mirrors; 5. The fluorescence reader of claim 4.

8. the plurality of reflectors include only one of the convex mirrors, one of the concave mirrors, and one retroreflector; 5. The fluorescence reader of claim 4.

9. the plurality of reflectors include only one of the convex mirrors, one of the concave mirrors, and one right-angle reflector; 5. The fluorescence reader of claim 4.

10. A fluorescence reading method for acquiring fluorescence from a microarray, comprising: The fluorescence emitted from the microarray is reflected multiple times by multiple reflectors in a light receiving optical system to form an image on an image forming plane; receiving the fluorescent light with an optical sensor located on the imaging plane; A fluorescence reading method comprising:

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