Analytical devices and analytical methods

Abstract

To provide an analytical device and analytical method that enable the implementation of digital assays simply and with high measurement sensitivity. [Solution] This disclosure relates to an analytical device comprising: a device holder for holding a microchamber device; a light source for irradiating a first surface of the microchamber device with laser light from diagonally above or diagonally below; one or more aspherical lenses arranged such that their optical axes are substantially parallel to the normal of the first surface of the microchamber device; an imaging device for imaging the microchamber device through the aspherical lenses; and a first optical filter located between the microchamber device and the imaging device, the first optical filter blocking the laser light emitted by the light source and transmitting at least a portion of the light on the longer wavelength side than the laser light emitted by the light source.

Description

[Technical Field]

[0001] This disclosure relates to analytical devices and analytical methods, particularly analytical devices and analytical methods used for observing or measuring phenomena or reactions related to cell or biomolecular samples in the fields of biochemistry, cell biology, biophysics, and other research, as well as for observing and measuring other fine particles. [Background technology]

[0002] Digital assays are known as a measurement method that can simultaneously detect hundreds of thousands of molecules. In digital assays, the target molecule is confined to a microscopic space, enabling quantitative measurement at the single-molecule level.

[0003] Patent documents 1 to 7 and non-patent documents 1 to 2 disclose methods for performing digital assays using a microchamber device equipped with wells having a volume of approximately fL. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] Patent No. 3727026 [Patent Document 2] Japanese Patent Publication No. 2018-38384 [Patent Document 3] Japanese Patent Publication No. 2022-31760 [Patent Document 4] International Publication No. 2017 / 200070 [Patent Document 5] International Publication No. 2018 / 181488 [Patent Document 6] International Publication No. 2019 / 168200 [Patent Document 7] International Publication No. 2020 / 179858 [Non-patent literature]

[0005] [Non-Patent Document 1] KV Tabata et al., Sci.Rep., 2019, 9, 1067 [Non-Patent Document 2] Y. Minagawa et al., Lab on a Chip, 2019, 19, 2678 [Overview of the project] [Problems that the invention aims to solve]

[0006] In the aforementioned patent and non-patent documents, a device that can easily and accurately perform digital assays has not yet been realized. For example, while non-patent document 2 discloses a device for digital assays, there is a need for a device that can perform digital assays more easily or with higher accuracy.

[0007] Therefore, the present invention aims to provide an analytical device and analytical method that can implement a digital assay simply and with high measurement sensitivity. [Means for solving the problem]

[0008] An analytical device according to one embodiment of the present invention comprises: a device holder for holding a microchamber device; a light source for irradiating a first surface of the microchamber device with laser light from diagonally above or diagonally below; one or more aspherical lenses arranged such that their optical axes are substantially parallel to the normal of the first surface of the microchamber device; an imaging device for imaging the microchamber device through the aspherical lenses; and a first optical filter located between the microchamber device and the imaging device, which blocks the laser light emitted by the light source and transmits at least a portion of the light on the longer wavelength side than the laser light emitted by the light source.

[0009] In the above-described analytical device, the light source irradiates the first surface of the microchamber device with laser light from diagonally above or diagonally below. Because laser light tends to have high light intensity and directivity, it is possible to irradiate the analyte-containing sample with high intensity while suppressing stray light that becomes background noise in the analysis. Furthermore, by irradiating the first surface with laser light from diagonally above or diagonally below, that is, by setting the incident angle of the laser light to more than 0° and less than 90°, the signal intensity from the analyte-containing sample introduced into the microchamber device can be improved, and the measurement sensitivity can be improved. One reason for this is presumed to be as follows: That is, by adopting the above configuration, compared to the case where the incident angle of the laser light is 90° and the excitation light is irradiated onto the microchamber device as evanescent light, the light intensity that passes through the microchamber device and irradiates the analyte-containing sample can be increased, and as a result, the signal intensity is presumed to be improved. Furthermore, compared to the case where the excitation light is irradiated with a laser beam incidence angle of 0°, it is possible to suppress the incidence of light reflected at the interface between the microchamber device and the atmosphere, and light transmitted through the microchamber device, into the imaging device. This reduces background light and is expected to improve measurement sensitivity. In addition, since one or more aspherical lenses are placed between the imaging device and the device holder, it is possible to maintain image flatness and improve the spatial resolution of the imaging device while suppressing the occurrence of chromatic aberration. As a result, the above analytical device enables the implementation of digital assays simply and with high measurement sensitivity.

[0010] In the analytical device described above, it is preferable to include a focusing lens that concentrates the laser light emitted from the light source. In this case, the laser light emitted from the light source is the laser light that has passed through the focusing lens and been concentrated. By including such a focusing lens, the excitation light intensity can be improved more efficiently, the signal intensity from the analyte-containing sample set in the microchamber device is improved, and the measurement sensitivity of the digital assay tends to improve.

[0011] In the above analysis device, it is preferable that the magnitude of the angle formed by the first surface of the microchamber device and the central axis of the light beam of the laser light irradiated on the microchamber device is 3 to 40°. Thereby, the excitation light intensity is improved, the signal intensity from the analyte-containing sample set in the microchamber device is further improved, and the measurement sensitivity of the digital assay tends to be improved.

[0012] In the above analysis device, a plurality of aspherical lenses may be provided. The plurality of aspherical lenses are, in order from the microchamber device side, a first lens having a positive refractive power, a second lens having a negative refractive power, and a third lens having a positive refractive power, and both surfaces of the first lens, the second lens, and the third lens are aspherical, and it is preferable to satisfy the following formulas (1), (2), (3), (4), (5) and (6). (1) 4.30 ≤ f1 / f ≤ 4.95 (2) 0.120 ≤ φ1 / n1 ≤ 0.160 (3) 0.0030 ≤ φ1 / ν1 ≤ 0.0050 (4) -3.20 ≤ f2 / f ≤ -2.70 (5) -0.240 ≤ φ2 / n2 ≤ -0.160 (6) -0.0200 ≤ φ2 / ν2 ≤ -0.0120 f: Overall focal length of the attachment lens with respect to the d-line f1: Focal length of the first lens with respect to the d-line φ1: Product of the power of the first lens with respect to the d-line and f (f / f1) n1: Refractive index of the first lens with respect to the d-line ν1: Abbe number of the first lens f2: Focal length of the second lens with respect to the d-line φ2: Product of the power of the second lens with respect to the d-line and f (f / f2) n2: Refractive index of the second lens with respect to the d-line ν2: Abbe number of the second lens

[0013] The above-mentioned aspherical lens satisfies equations (1) and (4) while also satisfying equations (2) and (5), thereby further suppressing the deterioration of image flatness obtained by the imaging device. Furthermore, by satisfying equations (3) and (6) while also satisfying equations (1) and (4), chromatic aberration in the image obtained by the imaging device can be suppressed.

[0014] In the above-described analysis device, the multiple aspherical lenses may satisfy the following equations (I) to (VI) instead of equations (1) to (6) above. According to such an embodiment, deterioration of the flatness of the image obtained by the imaging device is suppressed, chromatic aberration is suppressed, and in addition, a wide area can be observed. (I)-10.0≦f1 / f≦8.00 (II)-0.1500≦φ1 / ν1≦0.0500 (III) 0.100 ≤ φ2 / n2 ≤ 2.000 (IV) -80.0 ≤ f3 / f ≤ 30.0 (V)-0.300≦φ3 / n3≦0.900 (VI)-0.0200≦φ3 / ν3≦0.0500 f: Total focal length of the attachment lens relative to the d line f1: Focal length of the first lens relative to the d line φ1: The product of the power of the first lens with respect to the d line and f (f / f1) ν1: Abbe number of the first lens f2: Focal length of the second lens relative to the d line φ2: The product of the power of the second lens with respect to the d line and f (f / f2) n2: Refractive index of the second lens with respect to the d line f3: Focal length of the third lens relative to the d line φ3: The product of the power of the third lens on the d line and f (f / f3) n3: Refractive index of the third lens with respect to the d line ν3: Abbe number of the third lens

[0015] In the analytical device described above, preferably, the aspherical lens is made of a non-fluorescent material. According to this embodiment, in the analytical device, it is possible to prevent the lens itself from emitting fluorescence due to light from a light source or light emitted from an analyte-containing sample in the microchamber device, thereby preventing noise from being generated during measurement.

[0016] In the above-described analysis device, preferably, the device holder or aspherical lens is movable in the direction normal to the first surface of the microchamber device. In this embodiment, the device holder can be moved closer to or further away from the imaging device, making it easier to focus the imaging device on the observation area in the device holder.

[0017] In the above-described analysis device, a plurality of wells may be provided on the first surface of the microchamber device. In this embodiment, an information processing device connected to or integrated with the imaging device may be further provided, and the information processing device may detect among the plurality of imaged wells that emit light in response to light from a light source.

[0018] An analytical method according to one embodiment of the present invention includes introducing an analyte-containing sample into the microchamber device of the analytical device described above, irradiating the sample with excitation light corresponding to the compound contained in the sample from a light source, and detecting the emission originating from the compound excited by the excitation light by imaging the microchamber device with an imaging device. Because the above analytical method is performed using the above analytical device, a digital assay can be realized simply and with high measurement sensitivity.

[0019] In the above analytical method, it is preferable that the analyte-containing sample contains a fluorescent dye or quantum dots. The above analytical method can perform quantitative analysis of the substance labeled by the above substance at the single-molecule level. This is because the above analytical method has high measurement sensitivity.

[0020] In the above analysis method, preferably, the first optical filter blocks light having a wavelength corresponding to the excitation light of the compound and transmits light having a wavelength corresponding to the light emitted by the excited compound. According to this embodiment, the signal-to-noise ratio can be further improved because stray light, which is background noise in the analysis, is suppressed while light originating from the compound, which is the signal, is transmitted.

[0021] The above-described analysis device may include, instead of an imaging device, an imaging device holder for holding the imaging device. [Effects of the Invention]

[0022] According to the present invention, it is possible to provide an analytical device and analytical method that can easily and with high measurement sensitivity realize a digital assay. [Brief explanation of the drawing]

[0023] [Figure 1A] This is a schematic cross-sectional view showing an example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and the light generated and detected from the analyte-containing sample. [Figure 1B] This is an enlarged view of a microchamber device in an example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto the analyte-containing sample. [Figure 2] This is a schematic cross-sectional view showing an example of an optical system including an aspherical lens in the analytical device of this embodiment. [Figure 3] This is a schematic cross-sectional view showing another example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and generated and detected from the analyte-containing sample. [Figure 4] This is a schematic cross-sectional view showing yet another example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and generated and detected from the analyte-containing sample. [Figure 5]This is a schematic cross-sectional view showing yet another example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and generated and detected from the analyte-containing sample. [Figure 6] This is a schematic cross-sectional view showing yet another example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and generated and detected from the analyte-containing sample. [Figure 7] This is a schematic cross-sectional view showing yet another example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and generated and detected from the analyte-containing sample. [Figure 8] This is a schematic cross-sectional view showing yet another example of the analytical device of this embodiment. The dashed arrows indicate the optical path of light irradiated onto an analyte-containing sample and generated and detected from the analyte-containing sample. [Figure 9] This is a schematic cross-sectional view showing an example of introducing an analyte-containing sample into a microchamber device in the analytical device of this embodiment. [Figure 10] This figure shows the surfaces R1 to R11 and the thickness (distance) d1 to d10 in the numerical example. [Figure 11] This is a schematic cross-sectional view showing the optical system of numerical example 6. [Figure 12] This is a schematic cross-sectional view showing the optical system of numerical example 7. [Figure 13] This is a schematic cross-sectional view showing the optical system of numerical example 8. [Figure 14] This is a schematic cross-sectional view showing the optical system of numerical example 9. [Figure 15] This is a schematic cross-sectional view showing the optical system of numerical example 10. [Figure 16] This is a schematic cross-sectional view showing the optical system of numerical example 11. [Figure 17] This figure shows the results for Examples 1-13 and Comparative Example 1. [Figure 18] This figure shows the results for Example 14 and Comparative Example 2. [Modes for carrying out the invention]

[0024] The following description will detail embodiments for carrying out the present invention (hereinafter referred to as "this embodiment") with reference to the drawings, but the present invention is not limited thereto, and various modifications are possible without departing from its essence. In the following drawings, identical or similar parts are denoted by the same or similar reference numerals. The drawings are schematic and do not necessarily correspond to actual dimensions or proportions. There may be parts in the drawings where the relationships between dimensions or proportions differ from those of other parts.

[0025] [First Embodiment] (Analysis device) Figure 1A is a schematic cross-sectional view of the analysis device 10 of this embodiment. As shown in Figure 1A, the analysis device 10 includes a device holder 320 for holding a microchamber device 300, a light source 500 for irradiating the microchamber device 300 with laser light L, one or more aspherical lenses 100 arranged so that their optical axes are substantially parallel to the normal to the first surface of the microchamber device 300, an imaging device 400 for imaging the microchamber device 300 through the aspherical lenses 100, a first optical filter 200 located between the microchamber device 300 and the imaging device 400, and a focusing lens 510 for focusing the laser light L irradiated from the light source 500. Here, the first optical filter 200 is an optical filter that blocks the light emitted by the light source 500 and transmits at least some of the light on the longer wavelength side than the light emitted by the light source. The imaging device 400 images the microchamber device 300 through the aspherical lenses 100 using the imaging lens 420. As shown in Figure 6 and other figures described later, the condensing lens 510 may be omitted.

[0026] In the embodiment shown in Figure 1A, the light source 500 irradiates the microchamber device 300 with laser light L from diagonally above or diagonally below the first surface of the microchamber device 300. Since laser light tends to have high light intensity, it is possible to irradiate the analyte-containing sample with high intensity excitation light. Also, since laser light tends to have high directivity, it is easy to irradiate only a specific area with light, and prevent other areas from being irradiated with light. Therefore, stray light that becomes background noise in the analysis can be suppressed. Furthermore, by irradiating the microchamber device 300 with laser light L from diagonally above or diagonally below the first surface, the signal intensity from the analyte-containing sample set in the microchamber device can be improved. The reason for this is not particularly limited, but is presumed to be as follows. Irradiating the microchamber device 300 with laser light L from diagonally above or diagonally below the first surface means that the angle of incidence of the laser light L to the microchamber device 300 is greater than 0° and less than 90°. Here, by making the incident angle of the laser light L smaller than 90°, the light intensity irradiated onto the analyte-containing sample after passing through the microchamber device 300 can be increased compared to when the excitation light is irradiated onto the microchamber device 300 as evanescent light. In addition, by making the incident angle of the laser light L onto the microchamber device 300 greater than 0°, it is possible to suppress the incidence of light reflected at the interface between the microchamber device 300 and the atmosphere, and light transmitted through the microchamber device, onto the imaging device 400. Therefore, it is thought that the signal intensity from the analyte-containing sample set in the microchamber device can be improved, and the measurement sensitivity can be improved. Furthermore, by positioning the light source 500 so that the laser light L is irradiated from diagonally above or diagonally below the first surface of the microchamber device 300, the observation optical system such as the imaging device 400, imaging lens 420, first optical filter 200, and aspherical lens 100 can be positioned perpendicularly above the first surface of the microchamber device 300, thus simplifying the device configuration.It is preferable that the light source 500 irradiates the microchamber device 300 with laser light L from the same side as the imaging device 400.

[0027] Furthermore, when a light source is said to irradiate the microchamber device with laser light from diagonally above or diagonally below the first surface of the microchamber device, it means that the angle between the central axis of the laser beam irradiated onto the microchamber device and the first surface of the microchamber device is greater than 0° and less than 90°. Therefore, this embodiment excludes the case in which an analyte-containing sample placed in the microchamber device is irradiated with excitation light solely by evanescent waves generated by light guided by total internal reflection within the microchamber device. In addition, it is not necessary for the light source to be positioned diagonally above or diagonally below the first surface of the microchamber device; a focusing lens as shown in Figure 1A or an optical element such as an optical fiber can be used to irradiate the first surface of the microchamber device with laser light from the light source from diagonally above or diagonally below.

[0028] The angle A between the first surface of the microchamber device 300 and the central axis of the laser beam L irradiated onto the microchamber device 300 is preferably 1 to 70°, 2 to 60°, 3 to 50°, 3 to 40°, or 3 to 30°. When the angle A is 1° or more, reflection at the interface between the microchamber device 300 and the atmosphere is suppressed, making it possible to irradiate the analyte-containing sample with more light, and tending to further improve the signal intensity from the analyte-containing sample. Also, when the angle A is 70° or less, reflection of the laser beam L at the interface between the microchamber device 300 and the atmosphere and incident on the imaging device can be further suppressed. From a similar viewpoint, the angle A may be 4 to 25°, 5 to 20°, or 6 to 15°. The angle A may be adjusted by changing the arrangement of the light source 500, the focusing lens 510, and / or the microchamber device 300.

[0029] Referring to Figure 1B, the incident angle B of the laser light L incident on the chamber within the microchamber device 300 will be described. Figure 1B is an enlarged view of the microchamber device 300 in an example of the analytical device of this embodiment. As shown in Figure 1B, the microchamber device 300 is prepared so that the main body portion 300a, which includes the microchamber, is sandwiched between a plate-shaped member 306 and a lid portion 310, which will be described later. The main body portion 300a may include, for example, a flow channel 312 and a well 302, as will be described later.

[0030] As shown in Figure 1B, after the laser beam L is incident on the microchamber device 300, the laser beam L is incident on the main body portion 300a, which includes the microchamber within the microchamber device 300, at an incident angle B. As shown in Figure 1B, the incident angle B is the angle between the normal to the surface of the main body portion 300a on the side of the plate-shaped member 306 and the central axis of the laser beam L. However, in Figure 1B, the plate-shaped member 306 and the lid portion 310 may be arranged in reverse. The incident angle B can be calculated using Snell's law based on the range of angle A described above, the material (e.g., glass) contained in the plate-shaped member 306 or the lid portion 310, and the refractive index of air.

[0031] The incident angle B is preferably 25-70°, 25-60°, or 25-50°. Alternatively, the incident angle B is preferably 25-40°, 30-45°, or 33-50°. By having the incident angle B within the above numerical range, the signal intensity from the analyte-containing sample can be further improved, and the reflection of the laser light L from the microchamber device 300 and its incidence into the imaging device can be further suppressed.

[0032] The angle A is measured based on the central axis of the laser beam L that directly irradiates the microchamber device 300. For example, in Figure 1A, the angle A is the size of the angle between the central axis of the laser beam L after refraction by the focusing lens 510 and the first surface of the microchamber device 300. The angle A can be adjusted by changing the tilt angle of the laser light source 500 that emits the laser light, and the angle A can also be measured by measuring this tilt angle. There are no particular limitations on the method of changing the tilt angle, but one example is to use a goniometric stage GN05 / M manufactured by THORLABS.

[0033] Referring again to Figure 1A, the focusing lens 510 is not particularly limited as long as it can focus the laser light L emitted from the light source 500 and irradiate the microchamber device 300 with the focused laser light L. The focusing lens 510 may be, for example, a convex lens. Alternatively, the focusing lens 510 may be, for example, a spherical lens or an aspherical lens. As for aspherical lenses that can be used as focusing lenses 510, they are not particularly limited as long as their focal length is suitable for the size of the analysis device 10 and they can focus the light to the desired position, but an example is THORLABS' C060TMD-A (focal length f=9.6mm). The focal length of the focusing lens 510 can be adjusted according to the size of the microchamber device 300.

[0034] The analytical device 10 is used to analyze an analyte-containing sample by irradiating the analyte-containing sample introduced into the microchamber device 300 with laser light L from the light source 500, and imaging the luminescence originating from compounds in the sample excited by the laser light L using the imaging device 400.

[0035] The microchamber device 300 is not particularly limited as long as it can contain a liquid analyte-containing sample and transmits the emission wavelength so that the emission from the analyte-containing sample can be detected by the imaging device 400. The wavelength range transmitted by the microchamber device 300 is not particularly limited, as the emission wavelength may change depending on the type of compound contained in the sample. For example, the microchamber device 300 may be transparent to visible light.

[0036] The microchamber device 300 may have multiple wells on its first surface. The volume of each well is not particularly limited, but is, for example, 10 aL to 100 nL, preferably 1.0 fL to 1.0 pL. The opening diameter or bottom diameter of each well may be, for example, 1.0 μm to 40 μm, and the depth may be about 400 nm to 5.0 μm. The spacing between each well may be 1.0 μm to 100 μm. The area of ​​the region on the first surface of the microchamber device 300 where multiple wells are provided is not particularly limited, for example, 1 mm 2 More than 10cm 2 The following may be the case: The first surface of the microchamber device 300 is, for example, the bottom surface or the top surface of the microchamber device 300.

[0037] The wells provided on the first surface of the microchamber device 300 may be formed separated from each other by side walls having, for example, hydrophobic upper surfaces. More specifically, as shown in Figure 9, which will be described in detail later, if the wells 302 are separated from each other by side walls 304 having hydrophobic upper surfaces 304a, then by introducing a hydrophilic solution, which is an analyte-containing sample, into the microchamber device 300, and then introducing a hydrophobic solution, the analyte-containing sample can be introduced into only a number of wells, and the analyte-containing sample can be divided into volumes corresponding to the volume of each well.

[0038] The first surface of the microchamber device 300 may comprise a plate-like member 306 and a side wall 304 having a hydrophobic upper surface 304a, as shown in Figure 9. In this embodiment, it is preferable that the plate-like member 306 has a hydrophilic surface. A "hydrophilic surface" refers to a surface whose affinity for hydrophilic solvents is higher than its affinity for hydrophobic solvents. The plate-like member may be made of a solid material, such as glass, silicon, or polymer resin.

[0039] The side wall 304 is a structure that separates each of the multiple wells 302, provided on the surface of the plate-like member 306, preferably on a hydrophilic surface. The side wall 304 has a hydrophobic upper surface 304a. Here, "hydrophobic" is used interchangeably with "lipophilic," meaning that its affinity for hydrophobic solvents is higher than its affinity for hydrophilic solvents.

[0040] The well 302 is a region enclosed by the side walls 304, with a portion of the surface of the plate-like member 306 serving as its bottom surface. Therefore, it is preferable that the bottom surface of the well 302 be hydrophilic. The shape of the region enclosed by the bottom surface and side walls of the well may be, for example, cylindrical, prismatic, or the like.

[0041] The microchamber device 300 is placed in the device holder 320 after or before introducing the analyte-containing sample. In Figure 1A, the microchamber device 300 may be positioned with its first surface facing the imaging device 400 (i.e., upward in Figure 1A). The microchamber device 300 may have a lid facing the first surface to prevent the analyte-containing sample inside the microchamber device 300 from leaking out when it is placed in the device holder 320 with its first surface facing upward. The configuration of the lid is not particularly limited. The microchamber device 300 may be positioned with a surface other than the first surface facing the imaging device 400, for example, with the surface facing the first surface facing the imaging device 400.

[0042] The lid portion 310 shown in Figure 9 is positioned opposite the first surface of the microchamber device 300, thereby forming a space within the microchamber device 300 consisting of a channel 312 and a well 302. The surface of the lid portion 310 that faces the first surface of the microchamber device 300 (i.e., the surface constituting the channel 312) may be hydrophobic. In this case, for example, if the lid portion 310 is hydrophobic, the upper and side surfaces of the side wall 304 formed on the first surface are hydrophobic, and the bottom surface of the well 302 is hydrophilic, it is preferable that a hydrophilic solution is introduced only into the well 302. Examples of the microchamber device 300 include the device described in the examples and the device described in KV Tabata et al., Sci.Rep., 2019, 9, 1067.

[0043] The device holder 320 is a component for holding the microchamber device 300 in a predetermined position. The device holder 320 may be fixed or movable, but it is preferable that it be movable in the direction normal to the first surface of the microchamber device 300, i.e., in the up and down direction in Figure 1A. According to this embodiment, the device holder 320 can be moved closer to or further away from the imaging device 400, making it easy to focus the imaging device 400 on the observation area in the device holder 320.

[0044] The analyte-containing sample introduced into the microchamber device 300 is excited by laser light L emitted from the light source 500. The light source 500 is not particularly limited as long as it is capable of emitting laser light. The wavelength of the light emitted by the light source 500 can be appropriately changed depending on the type of compound contained in the sample. For example, the light source 500 may emit light at the wavelength of the absorption peak of the fluorescent substance contained in the sample. The laser light L emitted from the light source 500 is guided to the microchamber device 300 via the focusing lens 510.

[0045] The laser light L emitted from the light source 500 is irradiated onto the first surface of the microchamber device 300 from an oblique angle above or below. If the microchamber device 300 contains a substance that is excited by the light, the substance is excited by the irradiated laser light and generates light, typically fluorescence.

[0046] As described above, the substance contained in the sample within the microchamber device 300 emits light. The imaging device 400 detects the emitted light through the imaging lens 420, the aspherical lens 100, and the first optical filter 200. It is preferable that the analysis device 10 is measured in a state where ambient light is blocked, so that the imaging device 400 can easily detect the emitted light. For example, the analysis device 10 may be equipped with light-shielding means as shown in Figures 3 to 5 described later, or it may be installed in a darkroom.

[0047] The first optical filter 200 blocks the laser light L emitted by the light source 500 and transmits at least some of the light on the longer wavelength side than the laser light emitted by the light source 500. The transmission wavelength of the first optical filter 200 is selected to transmit the light emitted by the substances contained in the sample. Therefore, the first optical filter 200 exclusively blocks the laser light L emitted by the light source 500, which would be noise in the analysis, while exclusively transmitting the light emitted by the compounds contained in the sample.

[0048] The imaging device 400 images the microchamber device 300 via the imaging lens 420 and the aspherical lens 100, and detects light emission from the analyte-containing sample within the microchamber device 300. The imaging lens 420 may be an imaging lens attached to the imaging device 400. Examples of the imaging device 400 include microscopes, cameras, and information processing terminals such as smartphones and tablets. The details of the aspherical lens 100 will be described later, but it plays a role in zooming the imaging area of ​​the imaging device 400 and increasing the spatial resolution of the imaging device 400. By increasing the spatial resolution of the imaging device 400 with the aspherical lens 100, the overall optical performance, including the imaging lens attached to the imaging device, does not deteriorate, and minute objects can be observed with high resolution.

[0049] The imaging device 400 may have functions as an information processing device, such as a smartphone or tablet, or it may be connected to an information processing device. The imaging device 400 may transmit the captured data to the information processing device by wire or wireless connection and then perform information processing, or information processing may be performed by an information processing device integrated with the imaging device.

[0050] (Aspherical lenses) The analysis device 10 includes one or more aspherical lenses 100. The aspherical lenses 100 function as so-called attachment lenses that enhance the spatial resolution of the imaging device 400. Because the analysis device 10 includes aspherical lenses 100 as attachment lenses, it can improve the spatial resolution of the imaging device 400 while maintaining flatness and suppressing the occurrence of chromatic aberration. When spherical lenses are used as attachment lenses, the aberration balance is disrupted, and the optical performance of the entire optical system, including the imaging lens and the attachment lens, tends to deteriorate. The aspherical lenses 100 and / or the imaging device 400 may be movable in the direction normal to the first surface of the microchamber device 300, i.e., vertically in Figure 1A.

[0051] At least one aspherical lens 100 has a positive refractive power. This can improve the spatial resolution of the imaging device 400.

[0052] Figure 2 shows a schematic cross-sectional view of an example configuration of the aspherical lens 100. In Figure 2, the aspherical lens 100 consists of a first lens 101, a second lens 102, and a third lens 103. Figure 2 shows an example in which the aspherical lens 100 is mounted on an imaging lens 420 that images the microchamber device 300 via a first optical filter 200. In Figure 2, only the bottom of the microchamber device 300 is shown, with the side of the microchamber device 300 with the aspherical lens 100 being the outside of the microchamber device 300, and the side of the microchamber device 300 opposite the aspherical lens 100 being the inside of the microchamber device 300. The optical axes AX of the aspherical lens 100, the first optical filter 200, and the imaging lens 420 are arranged to substantially coincide with the observation area of ​​the microchamber device 300. The observation area is, for example, a plurality of wells provided on the first surface of the microchamber device 300. The multiple wells are located on the side opposite to the aspherical lens 100 in Figure 2.

[0053] In Figure 2, the aspherical lens 100 consists of a first lens 101 having a positive refractive power, a second lens 102 having a negative refractive power, and a third lens 103 having a positive refractive power, from the microchamber device 300 side. Both sides of the first lens 101, the second lens 102, and the third lens 103 are aspherical. The aspherical lens 100 preferably satisfies the following formulas (1), (2), (3), (4), (5), and (6).

[0054] (1) 4.30 ≤ f1 / f ≤ 4.95 (2) 0.120 ≤ φ1 / n1 ≤ 0.160 (3) 0.0030 ≤ φ1 / ν1 ≤ 0.0050 (4) -3.20 ≤ f² / f ≤ -2.70 (5) -0.240 ≤ φ2 / n2 ≤ -0.160 (6) - 0.0200 ≤ φ2 / ν2 ≤ - 0.0120 f: Focal length of the entire system with respect to the d - line of the attachment lens f1: Focal length of the first lens with respect to the d - line φ1: Product of the power of the first lens with respect to the d - line and f (f / f1) n1: Refractive index of the first lens with respect to the d - line ν1: Abbe number of the first lens f2: Focal length of the second lens with respect to the d - line φ2: Product of the power of the second lens with respect to the d - line and f (f / f2) n2: Refractive index of the second lens with respect to the d - line ν2: Abbe number of the second lens

[0055] Here, in this specification, the Abbe number is the value ν defined by the following formula using the refractive index n with respect to Fraunhofer's d - line (587.56 nm) d , refractive index n with respect to the F - line (486.1 nm) F , and refractive index n with respect to the C - line (656.3 nm) C is the value ν defined by the following formula d as follows ν d =(n d - 1) / (n F - n C )

[0056] Also, when a lens has a positive refractive power, it means that the power of the lens with respect to the d - line, that is, the reciprocal of the focal length of the lens with respect to the d - line is positive. When a lens has a negative refractive power, it means that the power of the lens with respect to the d - line, that is, the reciprocal of the focal length of the lens with respect to the d - line is negative

[0057] Equations (1) and (4) above define the ratios of the focal lengths of the first lens 101 and the second lens 102 to the total focal length of the aspherical lens 100, respectively. The ratio of the focal length of the third lens 103 to the total focal length of the aspherical lens 100 will be described later, but the aspherical lens 100 should be adjusted so that the desired total focal length f is obtained within the range that satisfies equations (1) and (4) above.

[0058] f1 / f may be 4.33 or greater, 4.34 or greater, 4.51 or greater, 4.62 or greater, or 4.90 or greater within the range of formula (1) above. Also, f1 / f may be 4.94 or less, 4.63 or less, 4.52 or less, or 4.35 or less within the range of formula (1) above.

[0059] f2 / f may be -3.18 or greater, -3.05 or greater, -3.01 or greater, -3.00 or greater, -2.88 or greater, or -2.76 or greater, within the range of equation (4) above. Also, f2 / f may be -2.80 or less, -2.87 or less, -3.00 or less, or -3.17 or less, within the range of equation (4) above.

[0060] The values ​​of f, f1, and f2 are not particularly limited, as they can be adjusted as appropriate by scaling, which is the process of changing the size of the optical system while maintaining the relationships between the components of the aspherical lens 100. For example, f may be between 1.10 mm and 10.0 mm, between 1.20 mm and 5.00 mm, or between 1.30 mm and 2.00 mm. Once f is set, f1 and f2 are designed to satisfy equations (1) and (4) above, respectively.

[0061] f should be adjusted according to the focal length of the imaging lens 420 and the desired magnification. The magnification of the attachment lens refers to the ratio of the image size with the attachment lens attached to the image size without the attachment lens. The magnification β is the focal length f of the imaging lens. 400 It is determined by the focal length f of the attachment lens, the distance between the imaging lens and the attachment lens, and other factors.

[0062] The magnification β of the aspherical lens 100 is preferably greater than 1.00, more preferably 1.10 or greater, and even more preferably 1.20 or greater. The focal length f of the aspherical lens 100 is equal to the focal length f of the imaging lens 420. 400 Depending on the value of β, you may adjust it as appropriate so that it falls within the above range.

[0063] Equations (2) and (5) above define the values ​​obtained by normalizing the ratio of power to refractive index of each lens in the first lens 101 and the second lens 102 by the power (1 / f) of the aspherical lens 100. In an optical system including multiple lenses, as the sum of the ratios of power to refractive index of each lens approaches 0, the flatness of the image plane of the entire optical system improves, that is, image field curvature is suppressed. The sum of the ratios of power to refractive index of each lens, or the value obtained by normalizing this sum by the power of the entire optical system, is also called the Petzval sum. In the aspherical lens 100, since φ1 / n1 and φ2 / n2 are defined by equations (2) and (5) above, the Petzval sum in the aspherical lens 100 can be brought closer to 0 by appropriately adjusting the value obtained by normalizing the ratio of power to refractive index of the third lens 103 by the power (1 / f) of the aspherical lens 100, thereby suppressing the deterioration of the flatness of the image obtained by the imaging device 400. Furthermore, φi (i=1,2,3) can also be understood as the ratio of the power of the i-th lens on the d-line (1 / fi) (i=1,2,3) to the total system power (1 / f) of aspherical lens 100 on the d-line.

[0064] φ1 / n1 may be 0.130 or greater, 0.140 or greater, 0.144 or greater, or 0.150 or greater, within the range of formula (2) above. Also, φ1 / n1 may be 0.155 or less, 0.145 or less, 0.142 or less, 0.140 or less, or 0.135 or less, within the range of formula (2) above.

[0065] φ2 / n2 may be -0.230 or greater, -0.220 or greater, -0.210 or greater, -0.205 or greater, or -0.190 or greater, within the range of formula (5) above. Also, φ2 / n2 may be -0.180 or less, -0.190 or less, -0.200 or less, or -0.210 or less, within the range of formula (5) above.

[0066] The value of n1 is not particularly limited as long as φ1 / n1 satisfies the above formula (2), but may be, for example, 1.20 or more and 2.00 or less, preferably 1.20 or more and 1.80 or less, more preferably 1.40 or more and 1.70 or less, and even more preferably 1.45 or more and 1.60 or less.

[0067] The value of n2 is not particularly limited as long as φ2 / n2 satisfies the above formula (5), but may be, for example, 1.30 or more and 2.10 or less, preferably 1.40 or more and 1.90 or less, more preferably 1.50 or more and 1.80 or less, and even more preferably 1.55 or more and 1.75 or less.

[0068] The relative magnitudes of n1 and n2 are not particularly limited, but it is preferable that n2 is larger, and more preferably that n2 is 0.05 or more, or 0.10 or more, larger than n1. In this case, the upper limit of the difference between n2 and n1 is not particularly limited and may be, for example, 0.50, 0.40, 0.30, or 0.20.

[0069] Equations (3) and (6) above define the values ​​obtained by normalizing the ratio of the power of the first lens 101 and the second lens 102 to the Abbe number using the power (1 / f) of the aspherical lens 100. In an optical system including multiple lenses, chromatic aberration can be suppressed in the entire optical system when the sum of the ratios of the power of each lens to the Abbe number approaches 0. In the aspherical lens 100, since φ1 / ν1 and φ2 / ν2 are defined by equations (3) and (6) above, the sum in the aspherical lens 100 can be brought closer to 0 by appropriately adjusting the value obtained by normalizing the ratio of the power of the third lens 103 to the Abbe number using the power (1 / f) of the aspherical lens 100, thereby reducing chromatic aberration in the image obtained by the imaging device 400.

[0070] φ1 / ν1 may be 0.0035 or greater, 0.0038 or greater, 0.0039 or greater, or 0.0040 or greater within the range of formula (3) above. Also, φ1 / ν1 may be 0.0045 or less, 0.0042 or less, 0.0040 or less, or 0.0039 or less within the range of formula (3) above.

[0071] φ2 / ν2 may be -0.0190 or greater, -0.0185 or greater, -0.0170 or greater, -0.0160 or greater, or -0.0150 or greater within the range of formula (6) above. Also, φ2 / ν2 may be -0.0130 or less, -0.0140 or less, -0.0150 or less, or -0.0160 or less within the range of formula (6) above.

[0072] The value of ν1 is not particularly limited as long as φ1 / ν1 satisfies the above formula (3), but may be, for example, 20 or more and 100 or less, preferably 30 or more and 90 or less, and more preferably 40 or more and 70 or less.

[0073] The value of ν2 is not particularly limited as long as φ2 / ν2 satisfies the above formula (6), but may be, for example, 5.0 or more and 80 or less, preferably 10 or more and 60 or less, and more preferably 15 or more and 40 or less.

[0074] The relative magnitudes of ν1 and ν2 are not particularly limited, but it is preferable that ν1 is larger, and it is even more preferable that ν1 is 10 or more, or 20 or more, larger than ν2. In this case, the upper limit of the difference between ν1 and ν2 is not particularly limited and may be, for example, 100, 80, 60, 50, or 40.

[0075] The aspherical lens 100 is preferably further satisfied with the following formulas (7) and (8).

[0076] (7) 0.000 ≤ P ≤ 0.510 (8) - 0.0046 ≤ Q ≤ 0.0046 P: Sum of φ1 / n1, φ2 / n2, and φ3 / n3 Q: The sum of φ1 / ν1, φ2 / ν2, and φ3 / ν3 φ3: The product of the power of the third lens on the d line and f (f / f3) f3: Focal length of the third lens relative to the d line n3: Refractive index of the third lens with respect to the d line ν3: Abbe number of the third lens

[0077] In equation (7) above, P is the value corresponding to the Petzval sum. In the aspherical lens 100, the first lens 101, the second lens 102, and the third lens 103 are aspherical lenses, so when the first lens 101 and the second lens satisfy equations (2) and (5) above, high flatness can be achieved in the image obtained by the imaging device 400. When P satisfies equation (7) above, even higher flatness can be achieved in the image obtained by the imaging device 400.

[0078] P may be 0.050 or more, 0.100 or more, 0.200 or more, 0.300 or more, or 0.400 or more within the range of formula (7) above. P is preferably 0.500 or less, more preferably 0.490 or less, and even more preferably 0.482 or less within the range of formula (7) above.

[0079] In equation (8) above, Q is a value that mainly relates to the chromatic aberration of the image obtained by the imaging device 400. In the aspherical lens 100, the first lens 101, the second lens 102, and the third lens 103 are aspherical lenses, so if the first lens 101 and the second lens satisfy equations (3) and (6) above, respectively, the occurrence of chromatic aberration in the image obtained by the imaging device 400 can be further suppressed. By satisfying equation (8) above, the occurrence of chromatic aberration in the image obtained by the imaging device 400 can be further reduced.

[0080] Q is preferably -0.0040 or greater, more preferably -0.0030 or greater, even more preferably -0.0010 or greater, and even more preferably 0.0000 or greater, within the range of formula (8) above. Q is preferably 0.0042 or less or 0.0040 or less, within the range of formula (8) above. Q may be 0.0038 or less, or 0.0035 or less.

[0081] The aspherical lens 100 may satisfy the following formulas (2A), (3A), (5A), (6A), (7A), and (8A) instead of the above formulas (2), (3), (5), (6), (7), and (8). (2A) 0.080 ≤ 1 / f1n1 ≤ 0.110 (3A) 0.0020 ≤ 1 / f1ν1 ≤ 0.0035 (5A)-0.160≦1 / f2n2≦-0.110 (6A)-0.0140≦1 / f2ν2≦-0.0080 (7A) 0.000 ≤ P' ≤ 0.350 (8A)-0.0031≦Q'≦0.0031 P': Sum of 1 / f1n1, 1 / f2n2, and 1 / f3n3 Q': Sum of 1 / f1ν1, 1 / f2ν2, and 1 / f3ν3

[0082] 1 / f1n1 may be 0.090 or greater, 0.094 or greater, 0.095 or greater, or 0.098 or greater, within the range of formula (2A) above. Also, 1 / f1n1 may be 0.105 or less, 0.98 or less, 0.095 or less, 0.093 or less, or 0.090 or less, within the range of formula (2A) above.

[0083] 1 / f1ν1 may be 0.0023 or greater, 0.0025 or greater, 0.0026 or greater, or 0.0027 or greater within the range of formula (3A) above. Also, 1 / f1ν1 may be 0.0030 or less, 0.0028 or less, 0.0026 or less, or 0.0025 or less within the range of formula (3A) above.

[0084] 1 / f2n2 may be -0.150 or greater, -0.142 or greater, -0.140 or greater, -0.130 or greater, or -0.127 or greater, within the range of equation (5A) above. Also, 1 / f2n2 may be -0.120 or less, -0.125 or less, -0.135 or less, or -0.140 or less, within the range of equation (5A) above.

[0085] 1 / f²ν² may be -0.0125 or greater, -0.0110 or greater, -0.0105 or greater, or -0.0100 or greater, within the range of equation (6A) above. Also, 1 / f²ν² may be -0.0085 or less, -0.0090 or less, -0.0100 or less, or -0.0105 or less, within the range of equation (6A) above.

[0086] P' and Q' are values ​​obtained by dividing P and Q by f, respectively. P' may be 0.030 or more, 0.070 or more, 0.130 or more, 0.200 or more, or 0.250 or more within the range of formula (7A) above. P' is preferably 0.340 or less, more preferably 0.330 or less, and even more preferably 0.320 or less within the range of formula (7A) above. Q' is preferably -0.0028 or greater, more preferably -0.0020 or greater, even more preferably -0.0010 or greater, and even more preferably 0.0000 or greater, within the range of formula (8A) above. Q' is preferably 0.0029 or less or 0.0028 or less, within the range of formula (8A) above. Q' may be 0.0025 or less, or 0.0023 or less.

[0087] The aspherical lens 100 preferably further satisfies the following formulas (9), (10), and (11). The aspherical lens 100 preferably satisfies the following formulas (9), (10), and (11) in addition to the above formulas (1) to (6), and more preferably satisfies the following formulas (9), (10), and (11) in addition to the above formulas (1) to (8).

[0088] (9) 1.10 ≤ f3 / f ≤ 1.40 (10) 0.000 ≤ φ3 / n3 ≤ 0.600 (11) 0.0000≦φ3 / ν3≦0.0200 The definitions of f3, φ3, n3, and ν3 are as described above.

[0089] Equations (9) to (11) above define the characteristics of the third lens 103. Since the first lens 101 and the second lens 102 of the aspherical lens 100 satisfy equations (1) to (6) above, even when mounted on an existing imaging lens, the optical performance of the image obtained by the imaging device 400 does not deteriorate, and fine objects can be observed with high resolution. By appropriately adjusting the characteristics of the third lens 103, the desired characteristics can be achieved in the aspherical lens 100, but by having the third lens 103 satisfy equations (9) to (11) above, the deterioration of the flatness of the image obtained by the imaging device 400 and the occurrence of chromatic aberration can be further suppressed.

[0090] Equation (9) above defines the ratio of the focal length of the third lens 103 to the total focal length of the aspherical lens 100.

[0091] f3 / f may be 1.15 or greater, 1.20 or greater, or 1.24 or greater within the range of formula (9) above. Also, f3 / f may be 1.35 or less, 1.30 or less, or 1.28 or less within the range of formula (9) above. The value of f3 is not particularly limited, as it can be appropriately adjusted by scaling the aspherical lens 100.

[0092] Equation (10) above defines the value obtained by normalizing the ratio of the power of the third lens 103 to the refractive index with the power (1 / f) of the aspherical lens 100.

[0093] φ3 / n3 may be 0.100 or more, 0.300 or more, 0.400 or more, 0.450 or more, or 0.500 or more, within the range of formula (10) above. Also, φ3 / n3 may be 0.580 or less, 0.550 or less, or 0.530 or less, within the range of formula (10) above.

[0094] The value of n3 is not particularly limited, but may be, for example, 1.20 or more and 2.00 or less, preferably 1.20 or more and 1.80 or less, more preferably 1.40 or more and 1.70 or less, and even more preferably 1.45 or more and 1.60 or less. The relative sizes of n1, n2, and n3 are not particularly limited, but it is preferable that n3 is smaller than n2. n3 may be the same as n1, or may be in the range of n1 ± 0.10.

[0095] The above equation (11) defines the value obtained by normalizing the ratio of the power of the third lens 103 to the Abbe number with the power (1 / f) of the aspherical lens 100.

[0096] φ3 / ν3 may be 0.0050 or greater, 0.0100 or greater, or 0.0140 or greater within the range of formula (11) above. Also, φ3 / ν3 may be 0.0190 or less, 0.0180 or less, or 0.0150 or less within the range of formula (11) above.

[0097] The value of ν3 is not particularly limited, but may be, for example, 20 or more and 100 or less, preferably 30 or more and 90 or less, and more preferably 40 or more and 70 or less. The relative magnitudes of ν1, ν2, and ν3 are not particularly limited, but it is preferable that ν3 is greater than ν2. ν3 may be the same as ν1, and may be in the range of n1 ± 5.0.

[0098] The aspherical lens 100 may satisfy the following formulas (10A) and (11A) instead of the above formulas (10) and (11). (10A) 0.000 ≤ 1 / f3n3 ≤ 0.400 (11A)0.0000≦1 / f3ν3≦0.0140

[0099] 1 / f3n3 may be 0.060 or greater, 0.200 or greater, 0.250 or greater, 0.300 or greater, or 0.330 or greater within the range of formula (10A) above. Also, 1 / f3n3 may be 0.380 or less, 0.370 or less, or 0.360 or less within the range of formula (10A) above.

[0100] 1 / f3ν3 may be 0.0030 or greater, 0.0070 or greater, or 0.0090 or greater within the range of formula (11A) above. Also, 1 / f3ν3 may be 0.0130 or less, 0.0120 or less, or 0.0100 or less within the range of formula (11A) above.

[0101] The materials constituting the first lens 101, the second lens 102, and the third lens 103 are not particularly limited and may be inorganic materials such as glass, organic materials such as resin, or composite materials combining inorganic and organic materials.

[0102] The first lens 101, the second lens 102, and the third lens 103 are preferably made of a non-fluorescent material. In this embodiment, in the analytical device 10, it is possible to prevent the lenses themselves from emitting fluorescence due to light from the light source 500 or light emitted from the analyte-containing sample in the microchamber device 300, thereby preventing noise from being generated during measurement.

[0103] In an embodiment in which the first lens 101, the second lens 102, and the third lens 103 are composed of a non-fluorescent material, the aspherical lens 100 preferably satisfies the following formulas (1'), (2'), (3'), (4'), (5'), and (6'), more preferably satisfies the following formulas (7') and (8'), or the following formulas (9'), (10'), and (11'), and even more preferably satisfies the following formulas (7'), (8'), (9'), (10'), and (11'). According to this embodiment, it is possible to prevent the lens itself from emitting light, and to further suppress deterioration of the flatness of the image obtained by the imaging device 400 and the occurrence of chromatic aberration.

[0104] (1') 4.33 ≤ f1 / f ≤ 4.94 (2') 0.130 ≤ φ1 / n1 ≤ 0.155 (3') 0.0035 ≤ φ1 / ν1 ≤ 0.0042 (4')-3.05≦f2 / f≦-2.70 (5')-0.230≦φ2 / n2≦-0.180 (6')-0.0190≦φ2 / ν2≦-0.0130 (7') 0.000 ≤ P ≤ 0.482 (8')-0.0040≦Q≦0.0040 (9') 1.20 ≤ f3 / f ≤ 1.30 (10') 0.400 ≤ φ3 / n3 ≤ 0.550 (11') 0.0100 ≤ φ3 / ν3 ≤ 0.0150 The definitions of f, f1, f2, f3, n1, n2, n3, ν1, ν2, ν3, φ1, φ2, φ3, P, and Q are the same as described above.

[0105] Furthermore, in this embodiment, the aspherical lens 100 may satisfy the following formulas (2A'), (3A'), (5A'), (6A'), (7A'), (8A'), (10A'), and (11A') instead of the above formulas (2'), (3'), (5'), (6'), (7'), (8'), (10'), and (11'). (2A') 0.085 ≤ 1 / f1n1 ≤ 0.105 (3A')0.0023≦1 / f1ν1≦0.0028 (5A')-0.150≦1 / f2n2≦-0.140 (6A')-0.0105≦1 / f2ν2≦-0.0090 (7A')0.000≦P'≦0.320 (8A')-0.0028≦Q'≦0.0028 (10A')0.300≦1 / f3n3≦0.370 (11A')0.0070≦1 / f3ν3≦0.0100 The definitions of P' and Q' are the same as described above.

[0106] In equations (1') to (11') above, the values ​​of f1 / f, φ1 / n1, φ1 / ν1, f2 / f, φ2 / n2, φ2 / ν2, P, Q, f3 / f, φ3 / n3, φ3 / ν3, f, n1, n2, n3, ν1, ν2, and ν3 may be within the range of equations (1') to (11') above, obtained by arbitrarily combining the upper and lower limits exemplified in equations (1) to (11) above. Furthermore, in the above equations (2A'), (3A'), (5A'), (6A'), (7A'), (8A'), (10A'), and (11A'), the values ​​of 1 / f1n1, 1 / f1ν1, 1 / f2n2, 1 / f2ν2, P', Q', 1 / f3n3, and 1 / f3ν3 may be within the range obtained by arbitrarily combining the upper and lower limits exemplified in the above equations (2A), (3A), (5A), (6A), (7A), (8A), (10A), and (11A).

[0107] The above formulas (1') to (11'), as well as (2A'), (3A'), (5A'), (6A'), (7A'), (8A'), (10A'), and (11A'), are formulas that further restrict the numerical range of the above formulas (1) to (11), as well as (2A), (3A), (5A), (6A), (7A), (8A), (10A), and (11A). When the first lens 101, the second lens 102, and the third lens 103 are made of non-fluorescent material, the refractive index, Abbe number, and shape of each lens may be limited to some extent. By limiting the numerical range of equations (1) to (11) to the ranges of equations (1') to (11') and (2A'), (3A'), (5A'), (6A'), (7A'), (8A'), (10A'), and (11A'), the deterioration of the flatness of the image obtained by the imaging device 400 and the occurrence of chromatic aberration can be further suppressed.

[0108] In another embodiment, the aspherical lens 100 may consist of a first lens having positive or negative refractive power, a second lens having positive refractive power, and a third lens having positive or negative refractive power, viewed from the microchamber device 300 side. Both sides of the first lens 101, the second lens 102, and the third lens 103 are aspherical. It is also preferable that the aspherical lens 100 satisfies the following formulas (I), (II), (III), (IV), (V), and (VI).

[0109] (I)-10.0≦f1 / f≦8.00 (II)-0.1500≦φ1 / ν1≦0.0500 (III) 0.100 ≤ φ2 / n2 ≤ 2.000 (IV) -80.0 ≤ f3 / f ≤ 30.0 (V)-0.300≦φ3 / n3≦0.900 (VI)-0.0200≦φ3 / ν3≦0.0500 The definitions of f, f1, φ1, ν1, f2, φ2, n2, f3, φ3, n3, and ν3 are the same as above.

[0110] Equations (I) and (IV) above define the ratios of the focal lengths of the first lens 101 and the third lens 103 to the total focal length of the aspherical lens 100, respectively. The ratio of the focal length of the second lens 102 to the total focal length of the aspherical lens 100 will be described later, but it is sufficient to adjust the aspherical lens 100 so that the desired total focal length f is obtained within the range that satisfies equations (I) to (VI) above.

[0111] f1 / f may be -9.00 or greater, -8.50 or greater, -3.00 or greater, -2.30 or greater, -1.40 or greater, -0.50 or greater, 1.50 or greater, 2.80 or greater, or 5.00 or greater, within the range of formula (I) above. Also, f1 / f may be 6.00 or less, 5.60 or less, 5.00 or less, 3.20 or less, 0.50 or less, -0.20 or less, -0.40 or less, -1.00 or less, -1.50 or less, -2.10 or less, or -7.00 or less, within the range of formula (I) above.

[0112] f3 / f may be -70.0 or greater, -67.0 or greater, -30.0 or greater, -15.0 or greater, -7.20 or greater, -5.10 or greater, 0.50 or greater, 1.25 or greater, 10.0 or greater, 15.0 or greater, or 18.0 or greater, within the range of formula (IV) above. Also, f3 / f may be 25.0 or less, 20.0 or less, 10.0 or less, 3.00 or less, 2.00 or less, 1.50 or less, 1.05 or less, -3.00 or less, -7.00 or less, -25.0 or less, -60.0 or less, or -65.0 or less, within the range of formula (IV) above.

[0113] The values ​​of f, f1, and f3 are not particularly limited, as they can be adjusted as appropriate by scaling, which is the process of changing the size of the optical system while maintaining the relationships between the components of the aspherical lens 100. For example, f may be between 1.10 mm and 10.0 mm, between 1.50 mm and 8.00 mm, or between 2.00 mm and 5.00 mm. Once f is set, f1 and f3 are designed to satisfy equations (I) and (IV) above, respectively.

[0114] The magnification β of the aspherical lens 100 is preferably greater than 1.00, more preferably 1.10 or greater, and even more preferably 1.20 or greater. The focal length f of the aspherical lens 100 is equal to the focal length f of the imaging lens. 400 Depending on the values ​​of the above, β may be adjusted as appropriate so that it falls within the above range.

[0115] Equations (II) and (VI) above define the values ​​obtained by normalizing the ratio of the power of each lens to the Abbe number in the first lens 101 and the third lens 103 by the power (1 / f) of the aspherical lens 100.

[0116] φ1 / ν1 may be -0.1500 or greater, -0.0900 or greater, -0.0500 or greater, -0.0350 or greater, -0.0200 or greater, -0.0065 or greater, 0.0030 or greater, 0.0050 or greater, or 0.0090 or greater, within the range of formula (II) above. Also, φ1 / ν1 may be 0.0200 or less, 0.0150 or less, 0.0080 or less, 0.0060 or less, -0.0055 or less, -0.0150 or less, -0.0300 or less, -0.0700 or less, or -0.0800 or less, within the range of formula (II) above.

[0117] φ3 / ν3 may be -0.0120 or greater, -0.0090 or greater, -0.0060 or greater, -0.0010 or greater, 0.0010 or greater, 0.0020 or greater, 0.0080 or greater, 0.0120 or greater, or 0.0170 or greater, within the range of formula (VI) above. Also, φ3 / ν3 may be 0.0200 or less, 0.0180 or less, 0.0140 or less, 0.0040 or less, 0.0025 or less, -0.0005 or less, -0.0040 or less, -0.0055 or less, or -0.0080 or less, within the range of formula (VI) above.

[0118] The value of ν1 is not particularly limited as long as φ1 / ν1 satisfies the above formula (II), but for example it may be 10 or more and 100 or less, preferably 15 or more and 90 or less, more preferably 18 or more and 70 or less, and even more preferably 19 or more and 60 or less.

[0119] The value of ν3 is not particularly limited as long as φ3 / ν3 satisfies the above formula (VI), but for example it may be 10 or more and 100 or less, preferably 15 or more and 90 or less, more preferably 18 or more and 75 or less, even more preferably 19 or more and 70 or less, and even more preferably 22 or more and 60 or less.

[0120] The relative magnitudes of ν1 and ν3 are not particularly limited; ν1 may be larger, and ν3 may be larger. It is more preferable that the absolute value of the difference between ν1 and ν3 is 10 or greater, or 20 or greater. In this case, the upper limit of the absolute value of the difference between ν1 and ν3 is not particularly limited and may be, for example, 100, 80, 60, 50, or 40.

[0121] Equations (III) and (V) above specify the values ​​obtained by normalizing the ratio of the power to the refractive index of each lens in the second lens 102 and the third lens 103 by the power (1 / f) of the aspherical lens 100.

[0122] φ2 / n2 may be 0.150 or more, 0.200 or more, 0.380 or more, 0.500 or more, 0.600 or more, 0.800 or more, or 0.950 or more, within the range of formula (III) above. Also, φ2 / n2 may be 1.500 or less, 1.100 or less, 0.700 or less, 0.450 or less, 0.300 or less, or 0.250 or less, within the range of formula (III) above.

[0123] φ3 / n3 may be -0.250 or greater, -0.170 or greater, -0.120 or greater, -0.090 or greater, -0.030 or greater, -0.010 or greater, 0.010 or greater, 0.030 or greater, 0.200 or greater, 0.300 or greater, 0.480 or greater, or 0.620 or greater, within the range of formula (V) above. Also, φ3 / n3 may be 0.800 or less, 0.640 or less, 0.500 or less, 0.100 or less, 0.040 or less, -0.005 or less, -0.060 or less, -0.080 or less, or -0.110 or less, within the range of formula (V) above.

[0124] The value of n2 is not particularly limited as long as φ2 / n2 satisfies the above formula (III), but may be, for example, 1.20 or more and 2.00 or less, preferably 1.30 or more and 1.80 or less, more preferably 1.40 or more and 1.75 or less, even more preferably 1.45 or more and 1.75 or less, and even more preferably 1.50 or more and 1.70 or less.

[0125] The value of n3 is not particularly limited as long as φ3 / n3 satisfies the above formula (V), but may be, for example, 1.20 or more and 2.00 or less, preferably 1.30 or more and 1.80 or less, more preferably 1.40 or more and 1.75 or less, even more preferably 1.45 or more and 1.75 or less, and even more preferably 1.50 or more and 1.70 or less.

[0126] The relative sizes of n2 and n3 are not particularly limited, but n2 may be larger than n3. The absolute value of the difference between n2 and n3 may be greater than or equal to 0.03, 0.05, or 0.07. In this case, the upper limit of the absolute value of the difference between n2 and n3 is not particularly limited and may be, for example, 0.50, 0.40, 0.30, 0.20, or 0.10.

[0127] The aspherical lens 100 is preferably further satisfied with the following formulas (VII) and (VIII).

[0128] (VII) 0.000 ≤ P ≤ 0.800 (VIII)-0.0600≦Q≦0.0500 The definitions of P, Q, n1, and ν2 are the same as above.

[0129] In equation (VII) above, P is the value corresponding to the Petzval sum. In the aspherical lens 100, since the first lens 101, the second lens 102, and the third lens 103 are aspherical lenses, if the second lens 102 and the third lens 103 satisfy equations (III) and (V) above, respectively, high flatness can be achieved for the entire optical system. If P satisfies equation (VII) above, even higher flatness can be achieved for the entire optical system.

[0130] P may be 0.050 or more, 0.160 or more, 0.200 or more, 0.250 or more, 0.400 or more, 0.550 or more, 0.570 or more, or 0.650 or more, within the range of formula (VII) above. P is preferably 0.680 or less, but may also be 0.600 or less, 0.570 or less, 0.480 or less, 0.280 or less, or 0.230 or less, within the range of formula (VII) above.

[0131] In equation (VIII) above, Q is a value that mainly relates to the chromatic aberration of the aspherical lens 100. In the aspherical lens 100, the first lens 101, the second lens 102, and the third lens 103 are aspherical lenses, so if the first lens 101 and the third lens 103 satisfy equations (II) and (VI) above, respectively, the occurrence of chromatic aberration can be suppressed for the entire optical system. By satisfying equation (VIII) above, the occurrence of chromatic aberration can be further reduced for the entire optical system.

[0132] Q is preferably -0.0450 or more, more preferably -0.0300 or more, and may be -0.0110 or more, -0.0050 or more, 0.0100 or more, 0.0200 or more, or 0.0215 or more, within the range of formula (VIII) above. Q is preferably 0.0400 or less, more preferably 0.0250 or less, and may be 0.0220 or less, 0.0120 or less, -0.0030 or less, -0.0095 or less, -0.0280 or -0.0430 or less, within the range of formula (VII) above.

[0133] The aspherical lens 100 may satisfy the following formulas (II-A), (III-A), (VA), (VI-A), (VII-A), and (VIII-A) instead of the above formulas (II), (III), (V), (VI), (VII), and (VIII). (II-A) -0.0400≦1 / f1ν1≦0.0100 (III-A) 0.050≦1 / f2n2≦0.400 (VA) -0.100 ≤ 1 / f3n3 ≤ 0.300 (VI-A) -0.0080≦1 / f3ν3≦0.0100 (VII-A) 0.000≦P'≦0.650 (VIII-A)-0.0200≦Q'≦0.0200 The definitions of P' and Q' are the same as above.

[0134] 1 / f1ν1 may be -0.0260 or greater, -0.0110 or greater, -0.0090 or greater, -0.0020 or greater, or 0.0030 or greater within the range of formula (II-A) above. Also, 1 / f1ν1 may be 0.0040 or less, 0.0035 or less, -0.0010 or less, -0.0080 or less, -0.0100 or less, or -0.0240 or less within the range of formula (II-A) above.

[0135] 1 / f2n2 may be 0.100 or greater, 0.120 or greater, 0.160 or greater, 0.250 or greater, 0.280 or greater, or 0.300 or greater within the range of formula (III-A) above. Also, 1 / f2n2 may be 0.330 or less, 0.300 or less, 0.270 or less, 0.180 or less, 0.140 or less, or 0.120 or less within the range of formula (III-A) above.

[0136] 1 / f3n3 may be -0.060 or greater, -0.040 or greater, -0.010 or greater, 0.015 or greater, 0.130 or greater, or 0.190 or greater within the range of the above formula (VA). Also, 1 / f3n3 may be 0.210 or less, 0.150 or less, 0.030 or less, -0.001 or less, -0.030 or less, or -0.050 or less within the range of the above formula (VA).

[0137] 1 / f3ν3 may be -0.0042 or greater, -0.0025 or greater, -0.0005 or greater, 0.0010 or greater, 0.0040 or greater, or 0.0050 or greater within the range of equation (VI-A) above. Also, 1 / f3ν3 may be 0.0060 or less, 0.0045 or less, 0.0015 or less, -0.0001 or less, -0.0020 or less, or -0.0050 or less within the range of equation (VI-A) above.

[0138] P' may be 0.020 or more, 0.030 or more, 0.070 or more, 0.100 or more, 0.150 or more, 0.180 or more, or 0.200 or more, within the range of formula (VII-A) above. P' may be preferably 0.500 or less, more preferably 0.300 or less, and may also be 0.250 or less, 0.190 or less, 0.160 or less, or 0.110 or less, within the range of formula (VII-A) above. Q' is preferably -0.0140 or greater within the range of formula (VIII-A) above, but may also be -0.0050 or greater, -0.0010 or greater, 0.0000, 0.0030, or 0.0080 or greater. Q' is preferably 0.0140 or less within the range of formula (VIII-A) above, but may also be 0.0090 or less, 0.0040 or less, 0.0000 or less, or -0.0045 or less.

[0139] The aspherical lens 100 preferably further satisfies the following formulas (XI), (X), and (XI). In addition to the above formulas (I) to (VI), the aspherical lens 100 preferably satisfies the following formulas (IX), (X), and (XI), and more preferably satisfies the above formulas (I) to (VIII), in addition to the following formulas (IX), (X), and (XI).

[0140] (IX) -1.500≦φ1 / n1≦0.300 (X) 0.500 ≤ f² / f ≤ 3.50 (XI) 0.0050≦φ2 / ν2≦0.0400 The definitions of n1 and ν2 are as described above.

[0141] Since the aspherical lens 100 satisfies equations (I) to (VI) above, the overall optical performance does not deteriorate even when attached to an existing imaging lens, and fine objects can be observed with high resolution. If the aspherical lens 100 further satisfies equations (IX) to (XI) above, the deterioration of the flatness of the image obtained by the imaging device 400 and the occurrence of chromatic aberration can be further suppressed.

[0142] φ1 / n1 may be -1.260 or greater, -0.600 or greater, -0.500 or greater, -0.350 or greater, -0.280 or greater, -0.080 or greater, 0.050 or greater, 0.100 or greater, or 0.200 or greater, within the range of formula (IX) above. Also, φ1 / n1 may be 0.220 or less, 0.150 or less, 0.110 or less, -0.060 or less, -0.150 or less, -0.250 or less, -0.300 or less, -0.450 or less, -0.900 or less, or -1.200 or less, within the range of formula (IX) above.

[0143] The value of n1 is not particularly limited, but may be, for example, 1.10 or more and 2.00 or less, preferably 1.20 or more and 1.90 or less, more preferably 1.30 or more and 1.80 or less, and even more preferably 1.40 or more and 1.70 or less. The relative sizes of n1, n2, and n3 are not particularly limited, but n1 may be greater than n2, or n1 may be less than n2. The absolute value of the difference between n1 and n2 may be greater than 0.03, 0.05, or 0.07. In this case, the upper limit of the absolute value of the difference between n1 and n2 is not particularly limited, and may be, for example, 0.50, 0.40, 0.30, 0.20, or 0.15. n3 may be the same as n1, n1 may be greater than n3, or n1 may be less than n3.

[0144] f2 / f may be 0.600 or greater, 0.900 or greater, 1.00 or greater, 1.60 or greater, 2.00 or greater, or 2.70 or greater within the range of formula (X) above. Also, f2 / f may be 3.00 or less, 2.20 or less, 1.70 or less, 1.10 or less, 1.00 or less, or 0.700 or less within the range of formula (X) above. The value of f2 is not particularly limited as it can be appropriately adjusted by scaling the aspherical lens 100.

[0145] φ2 / ν2 may be 0.0100 or greater, 0.0140 or greater, 0.0160 or greater, 0.0170 or greater, 0.0180 or greater, 0.0200 or greater, or 0.0270 or greater, within the range of formula (XI) above. Also, φ2 / ν2 may be 0.0280 or less, 0.0200 or less, 0.0190 or less, 0.0180 or less, 0.0170 or less, 0.0155 or less, or 0.0120 or less, within the range of formula (XI) above.

[0146] The value of ν2 is not particularly limited, but may be, for example, 10 or more and 100 or less, preferably 15 or more and 80 or less, and more preferably 20 or more and 60 or less. The relative magnitudes of ν1, ν2, and ν3 are not particularly limited, but ν1 may be greater than ν2, or ν1 may be less than ν2. The absolute value of the difference between ν1 and ν2 is more preferably 10 or more, or 20 or more. In this case, the upper limit of the absolute value of the difference between ν1 and ν2 is not particularly limited, and may be, for example, 100, 80, 60, 50, or 40. ν3 may be the same as ν2, ν2 may be greater than ν3, or ν2 may be less than ν3.

[0147] The aspherical lens 100 may satisfy the following formulas (IX-A) and (XI-A) instead of the above formulas (IX) and (XI). (IX-A) -0.450≦1 / f1n1≦0.200 (XI-A) 0.0015≦1 / f2ν2≦0.0150

[0148] 1 / f1n1 may be -0.380 or greater, -0.160 or greater, -0.140 or greater, -0.030 or greater, 0.030 or greater, or 0.100 or greater within the range of equation (IX-A) above. Also, 1 / f1n1 may be 0.120 or less, 0.050 or less, -0.015 or less, -0.120 or less, -0.150 or less, or -0.370 or less within the range of equation (IX-A) above.

[0149] 1 / f²ν² may be 0.0028 or greater, 0.0035 or greater, 0.0045 or greater, 0.0070 or greater, 0.0080 or greater, or 0.0083 or greater within the range of equation (XI-A) above. Also, 1 / f²ν² may be 0.0090 or less, 0.0080 or less, 0.0075 or less, 0.0050 or less, or 0.0030 or less within the range of equation (XI-A) above.

[0150] In this embodiment, the materials constituting the first lens 101, the second lens 102, and the third lens 103 are not particularly limited and may be inorganic materials such as glass, organic materials such as resin, or composite materials combining inorganic and organic materials.

[0151] In this embodiment, it is preferable that the first lens 101, the second lens 102, and the third lens 103 are made of a non-fluorescent material. According to this embodiment, in the analytical device 10, it is possible to prevent the lenses themselves from emitting fluorescence due to light from the light source 500 or light emitted from the analyte-containing sample in the microchamber device 300, thereby preventing noise from being generated during measurement.

[0152] In this embodiment, when the first lens 101, the second lens 102, and the third lens 103 are made of a non-fluorescent material, the aspherical lens 100 preferably satisfies the following formulas (I'), (II'), (III'), (IV'), (V'), and (VI'), more preferably satisfies the following formulas (VII') and (VIII'), or the following formulas (IX'), (X'), and (XI'), and even more preferably satisfies the following formulas (VII'), (VIII'), (IX'), (X'), and (XI'). According to this embodiment, it is possible to prevent the lens itself from emitting light, and to further suppress deterioration of the flatness of the image obtained by the imaging device 400 and the occurrence of chromatic aberration.

[0153] (I') -8.50≦f1 / f≦5.60 (II') -0.0900≦φ1 / ν1≦0.0150 (III') 0.200 ≤ φ2 / n2 ≤ 1.100 (IV') -67.0≦f3 / f≦20.0 (V') -0.170≦φ3 / n3≦0.640 (VI') -0.0090≦φ3 / ν3≦0.0180 (VII') 0.000≦P≦0.680 (VIII') -0.0450≦Q≦0.0250 (IX') -1.260≦φ1 / n1≦0.220 (X') 0.630 ≤ f² / f ≤ 2.85 (XI') 0.0100≦φ2 / ν2≦0.0285 The definitions of f, f1, f2, f3, n1, n2, n3, ν1, ν2, ν3, φ1, φ2, φ3, P, and Q are the same as described above.

[0154] Furthermore, in this embodiment, the aspherical lens 100 may satisfy the following formulas (II-A'), (III-A'), (V-A'), (VI-A'), (VII-A'), (VIII-A'), (IX-A'), and (XI-A') instead of the above formulas (II'), (III'), (V'), (VI'), (VII'), (VIII'), (IX-A'), and (XI-A'). (II-A') -0.0260≦1 / f1ν1≦0.0040 (III-A') 0.100≦1 / f2n2≦0.330 (V-A') -0.060≦1 / f3n3≦0.210 (VI-A') -0.0042≦1 / f3ν3≦0.0060 (VII-A') 0.020≦P'≦0.500 (VIII-A') -0.0140≦Q'≦0.0140 (IX-A') -0.380≦1 / f1n1≦0.120 (XI-A') 0.0028≦1 / f2ν2≦0.0090 The definitions of P' and Q' are the same as described above.

[0155] In the above equations (I') to (XI'), the values ​​of f1 / f, φ1 / n1, φ1 / ν1, f2 / f, φ2 / n2, φ2 / ν2, P, Q, f3 / f, φ3 / n3, φ3 / ν3, f, n1, n2, n3, ν1, ν2, and ν3 may be within the range of the above equations (I') to (XI') and obtained by arbitrarily combining the upper and lower limits exemplified in the above equations (I) to (XI). Furthermore, in the above formulas (II-A'), (III-A'), (V-A'), (VI-A'), (VII-A'), (VIII-A'), (IX-A'), and (XI-A'), the values ​​of 1 / f1n1, 1 / f1ν1, 1 / f2n2, 1 / f2ν2, P', Q', 1 / f3n3, and 1 / f3ν3 may be within the range obtained by arbitrarily combining the upper and lower limits exemplified in the above formulas (II-A), (III-A), (VA), (VI-A), (VII-A), (VIII-A), (IX-A), and (XI-A).

[0156] The above formulas (I') to (XI'), as well as (II-A'), (III-A'), (V-A'), (VI-A'), (VII-A'), (VIII-A'), (IX-A'), and (XI-A') are formulas that further restrict the numerical range of the above formulas (I) to (XI), as well as (II-A), (III-A), (VA), (VI-A), (VII-A), (VIII-A), (IX-A), and (XI-A). When the first lens 101, the second lens 102, and the third lens 103 are made of non-fluorescent material, the refractive index, Abbe number, and shape of each lens may be limited to some extent. By limiting the numerical range of the above formulas (I) to (XI) to the ranges of the above formulas (I') to (XI') and (II-A'), (III-A'), (V-A'), (VI-A'), (VII-A'), (VIII-A'), (IX-A'), and (XI-A'), the deterioration of the flatness of the image obtained by the imaging device 400 and the occurrence of chromatic aberration can be further suppressed.

[0157] In one embodiment of the attachment lens of this embodiment, the above formulas (I) to (VI) and (IX) to (XI) may be the following formulas (I-1) to (VI-1) and (IX-1) to (XI-1). (I-1)-10.0≦f1 / f≦-7.00 (II-1)-0.0200≦φ1 / ν1≦-0.0055 (III-1) 0.600 ≤ φ2 / n2 ≤ 1.100 (IV-1)-80.0≦f3 / f≦-60.0 (V-1)-0.030≦φ3 / n3≦-0.005 (VI-1)-0.0060≦φ3 / ν3≦-0.0005 (IX-1)-0.080≦φ1 / n1≦-0.060 (X-1)0.900≦f2 / f≦1.10 (XI-1)0.0140≦φ2 / ν2≦0.0200

[0158] In one embodiment of the attachment lens of this embodiment, the above formulas (I) to (VI) and (IX) to (XI) may be the following formulas (I-2) to (VI-2) and (IX-2) to (XI-2). (I-2) 5.00 ≤ f1 / f ≤ 6.00 (II-2)0.0050≦φ1 / ν1≦0.0150 (III-2) 0.600 ≤ φ2 / n2 ≤ 0.700 (IV-2)-15.0≦f3 / f≦-3.00 (V-2)-0.120≦φ3 / n3≦-0.060 (VI-2)-0.0090≦φ3 / ν3≦-0.0040 (IX-2) 0.050 ≤ φ1 / n1 ≤ 0.150 (X-2)0.900≦f2 / f≦1.10 (XI-2)0.0160≦φ2 / ν2≦0.0200

[0159] In one embodiment of the attachment lens of this embodiment, the above formulas (I) to (VI) and (IX) to (XI) may be the following formulas (I-3) to (VI-3) and (IX-3) to (XI-3). (I-3) 1.50 ≤ f1 / f ≤ 5.00 (II-3)0.0030≦φ1 / ν1≦0.0080 (III-3) 0.150 ≤ φ2 / n2 ≤ 0.300 (IV-3) 15.0 ≤ f3 / f ≤ 25.0 (V-3)0.010≦φ3 / n3≦0.100 (VI-3)0.0010≦φ3 / ν3≦0.0040 (IX-3) 0.100 ≤ φ1 / n1 ≤ 0.300 (X-3)2.00≦f2 / f≦3.50 (XI-3)0.0100≦φ2 / ν2≦0.0190

[0160] In one embodiment of the attachment lens of this embodiment, the above formulas (I) to (VI) and (IX) to (XI) may be the following formulas (I-4) to (VI-4) and (IX-4) to (XI-4). (I-4)-3.00≦f1 / f≦-1.50 (II-4)-0.0350≦φ1 / ν1≦-0.0055 (III-4) 0.500 ≤ φ2 / n2 ≤ 0.700 (IV-4)-7.20≦f3 / f≦-3.00 (V-4)-0.170≦φ3 / n3≦-0.080 (VI-4)-0.0120≦φ3 / ν3≦-0.0055 (IX-4)-0.350≦φ1 / n1≦-0.150 (X-4)0.900≦f2 / f≦1.70 (XI-4)0.0050≦φ2 / ν2≦0.0200

[0161] In one embodiment of the attachment lens of this embodiment, the above formulas (I) to (VI) and (IX) to (XI) may be the following formulas (I-5) to (VI-5) and (IX-5) to (XI-5). (I-5)-2.30≦f1 / f≦-1.00 (II-5)-0.0500≦φ1 / ν1≦-0.0150 (III-5) 0.300 ≤ φ2 / n2 ≤ 0.640 (IV-5) 0.50 ≤ f3 / f ≤ 2.00 (V-5)0.480≦φ3 / n3≦0.800 (VI-5)0.0120≦φ3 / ν3≦0.0500 (IX-5)-0.600≦φ1 / n1≦-0.300 (X-5)1.00≦f2 / f≦2.20 (XI-5)0.0050≦φ2 / ν2≦0.170

[0162] In one embodiment of the attachment lens of this embodiment, the above formulas (I) to (VI) and (IX) to (XI) may be the following formulas (I-6) to (VI-6) and (IX-6) to (XI-6). (I-6)-1.40≦f1 / f≦-0.20 (II-6)-0.1500≦φ1 / ν1≦-0.0700 (III-6) 0.800 ≤ φ2 / n2 ≤ 1.500 (IV-6) 0.50 ≤ f3 / f ≤ 3.00 (V-6)0.300≦φ3 / n3≦0.640 (VI-6)0.0080≦φ3 / ν3≦0.0200 (IX-6)-1.500≦φ1 / n1≦-0.900 (X-6)0.500≦f2 / f≦1.00 (XI-6)0.0200≦φ2 / ν2≦0.0400

[0163] It goes without saying that this embodiment is not limited to the embodiments described above. In each of the above embodiments, the values ​​of f1 / f, φ1 / n1, φ1 / ν1, f2 / f, φ2 / n2, φ2 / ν2, f3 / f, φ3 / n3, φ3 / ν3, f, n1, n2, n3, ν1, ν2, and ν3 may be within the range of the above embodiments, obtained by arbitrarily combining the upper and lower limits exemplified in the above embodiments (I) to (XI).

[0164] The aspherical lens 100 of this embodiment enables observation of minute objects with a wider field of view. The half-angle of view ω of the aspherical lens 100 of this embodiment is preferably 15 degrees or more, more preferably 20 degrees or more, even more preferably 30 degrees or more, and may be 40 degrees or more, 45 degrees or more, 50 degrees or more, or 55 degrees or more. The upper limit of the half-angle of view ω is not particularly limited and may be, for example, 70 degrees, 65 degrees, or 60 degrees.

[0165] [Second Embodiment] Figure 3 is a schematic cross-sectional view of an analysis device 20 of this embodiment, which is different from the first embodiment. As shown in Figure 3, the analysis device 20 differs from the analysis device 10 shown in Figure 1A in that it includes a light-shielding means 600. As shown in Figure 3, the aspherical lens 100, the first optical filter 200, the device holder 320, the microchamber device 300, and the light source 500 may be housed in the light-shielding means 600. The light-shielding means 600 prevents a decrease in analysis accuracy due to stray light from the external environment. The light-shielding means is not particularly limited as long as it is a means to suppress light from entering the inside of the analysis device from the outside, or more specifically, a means to suppress or substantially prevent the detection of light other than the light that originates from the microchamber device and is detected by the imaging device via the first optical filter and the aspherical lens. Examples of such light-shielding means include light-shielding cases (dark boxes), light-shielding sheets, and light-shielding curtains. The light-shielding means may be made of resin, metal, wood, or cloth. The analysis device 20 may be installed in a darkroom.

[0166] The light-shielding means 600 only needs to suppress light of the same or near wavelength as the light generated from the microchamber device from entering the inside of the analysis device 20, but preferably it blocks visible light, and more preferably it blocks both visible light and ultraviolet light.

[0167] In Figure 3, the imaging device 400 is located outside the light-shielding means 600. The imaging device 400 has a hole 460 formed in it, and the imaging device 400 images the microchamber device 300 through the hole 460. In the light-shielding means 600, the hole 460 can be covered by the imaging lens 420, or the hole 460 can be closed by inserting the imaging lens 420 into the hole 460, thereby substantially preventing light from outside the light-shielding means 600 from being detected by the imaging lens 420. The light-shielding means 600 may also be provided with sealing means to seal the gap created by inserting the imaging lens 420 into the hole 460 or the gap created by covering the hole 460 with the imaging lens 420. Examples of sealing means include resin members (e.g., rubber rings) and adhesives. The imaging device 400 may also be located inside the light-shielding means 600.

[0168] [Third Embodiment] Figure 4 is a schematic cross-sectional view of an analysis device 30 of this embodiment, which is different from the first embodiment. As shown in Figure 4, the analysis device 30 differs from the analysis device 20 shown in Figure 3 in that it includes an imaging device holder 440 instead of an imaging device 400.

[0169] During analysis, the analysis device 30 is used by mounting the imaging device on the imaging device holder 440 such that the imaging lens is inserted into the hole 460 or covers the hole 460. Other configurations may be the same as in the first embodiment or its modified form.

[0170] [Fourth Embodiment] Figure 5 is a schematic cross-sectional view of an analysis device 40 of this embodiment, which is different from the first embodiment. As shown in Figure 5, the analysis device 40 differs from the analysis device 20 shown in Figure 3 in that it includes a light-shielding means 610 that houses a microchamber device 300, a device holder 320, an aspherical lens 100, and a first optical filter 200, instead of a light-shielding means 600.

[0171] In Figure 5, the light source 500 is located outside the light shielding means 610, and the focusing lens 510 is fitted into the light shielding means 610. Laser light emitted from the light source 500 reaches the focusing lens 510, is focused by the focusing lens 510, and irradiates the microchamber device 300. The periphery of the focusing lens 510 may be provided with a sealing means to seal the gap between it and the light shielding means 610. Examples of sealing means include resin components (e.g., rubber rings) and adhesives.

[0172] The condensing lens 510 may be installed on the outside or inside of the light-shielding means 610 so as to cover the holes provided in the light-shielding means 610. Other configurations may be the same as those of the first embodiment, the second embodiment, or their variations.

[0173] [Fifth Embodiment] Figure 6 is a schematic cross-sectional view of the analysis device 50 of this embodiment, which is different from the first embodiment. As shown in Figure 6, the analysis device 50 differs from the analysis device 20 shown in Figure 1A in that it does not have a focusing lens 510, and the laser light L emitted from the light source 500 is directly irradiated onto the microchamber device 300.

[0174] In the embodiment shown in Figure 6, angle A is the angle between the line connecting the laser beam emission port of the light source 500 and the spot on the first surface of the microchamber device 300 that is irradiated with laser light, and the first surface of the microchamber device 300.

[0175] In the fifth embodiment, an optical element for guiding the laser light L, such as an optical fiber or other waveguide, may be provided between the light source 500 and the microchamber device 300.

[0176] [Sixth Embodiment] Figure 7 is a schematic cross-sectional view of an analysis device 60 of this embodiment, which is different from the first embodiment. As shown in Figure 7, the analysis device 60 differs from the analysis device 10 shown in Figure 1A in that it further includes a lens 110. The lens 110 is provided between the first optical filter 200 and the microchamber device 300. By including the lens 110, the spatial resolution of the imaging device 400 tends to be further improved. In this embodiment, the lens 110 may be an aspherical lens. Alternatively, this embodiment may be realized using a microscope in which the lens 110 is the objective lens and the aspherical lens 100 is the eyepiece lens.

[0177] [Seventh Embodiment] Figure 8 is a schematic cross-sectional view of an analysis device 70 of this embodiment, which is different from the first embodiment. As shown in Figure 8, the analysis device 70 differs from the analysis device 50 shown in Figure 6 in that it includes a lens 110. The lens 110 is provided between the first optical filter 200 and the microchamber device 300. By including the lens 110, the spatial resolution of the imaging device 400 tends to be further improved. In this embodiment, the lens 110 may be an aspherical lens. Alternatively, this embodiment may be realized using a microscope in which the lens 110 is the objective lens and the aspherical lens 100 is the eyepiece lens.

[0178] [Differentiation] The above-described embodiment is illustrative for explaining the present invention and is not intended to limit the present invention to this embodiment alone. The present invention can be modified in various ways without departing from its spirit.

[0179] For example, in Figures 3 to 5, the light-shielding means 600 is configured to surround all four sides of each component and prevent stray light from entering except for the holes 460. However, the light-shielding means 600 does not necessarily have an upper surface, i.e., a surface or portion where the holes 460 are provided. In this embodiment, the imaging device 400 or the imaging device holder 440 covers the opening of the light-shielding means 600 to prevent stray light from entering the light-shielding means 600.

[0180] Therefore, the light-shielding means in the analytical device of this embodiment can house at least a microchamber device and a device holder, a first optical filter, and an aspherical lens, and has a hole in which the imaging lens of an imaging device can be positioned to image the microchamber device via the first optical filter and the aspherical lens; can house at least a microchamber device and a device holder, and a first optical filter, and has a hole in which the imaging lens of an imaging device can be positioned to image the microchamber device via the first optical filter and the aspherical lens, and an aspherical lens is provided in the hole; can house at least a microchamber device and a device holder, and has a hole in which the imaging lens of an imaging device can be positioned to image the microchamber device via the first optical filter and the aspherical lens Examples of the light-shielding means include: having a hole through which the imaging lens of an imaging device can be positioned to image a microchamber device via a casing, and an aspherical lens and a first optical filter provided in the hole; having a structure that can house at least a microchamber device and a device holder, a first optical filter and an aspherical lens, and is partially open, and is configured so that it can be sealed to prevent external light from entering the interior by installing an imaging device in the opening; and having a structure that can house at least a microchamber device and a device holder and a first optical filter, and is partially open, and is configured so that it can be sealed to prevent external light from entering the interior by installing an imaging device with an aspherical lens attached to the entire surface of the imaging lens in the opening. In these embodiments, at least one or more optical elements such as a light source and a focusing lens may be arranged inside the light-shielding means. The light-shielding means may be provided with an openable and closable outlet for removing each component housed inside. The ends of the opening may be provided with a sealing means such as a resin member (e.g., a rubber ring) and an adhesive.

[0181] Furthermore, in Figures 1A and 3-8, the image may be inverted vertically so that the imaging device 400 or the imaging device holder 440 is located at the bottom. In this case, the first surface of the microchamber device 300 is located at the bottom, and the lid may be omitted.

[0182] [Analysis method] The analytical device 10 shown in Figure 1A can perform analysis by introducing an analyte-containing sample into a microchamber device 300, irradiating the sample with excitation light corresponding to the compound contained in the sample from a light source 500, and detecting the luminescence originating from the compound excited by the excitation light by imaging the microchamber device 300 with an imaging device 400.

[0183] Therefore, the analysis method of this embodiment includes introducing an analyte-containing sample into a microchamber device of an analysis device, irradiating the sample with excitation light corresponding to the compound contained in the sample from a light source, and detecting the emission originating from the compound excited by the excitation light by imaging the microchamber device with an imaging device.

[0184] The analysis device 30 shown in Figure 4 has an imaging device placed in the imaging device holder 440, an analyte-containing sample is introduced into the microchamber device 300, excitation light corresponding to the compound contained in the sample is irradiated from the light source 500, and the microchamber device 300 is imaged by the imaging device. By detecting the emission originating from the compound excited by the excitation light, analysis can be performed.

[0185] Therefore, the analysis method of this embodiment may include arranging an imaging device in the imaging device holding section, introducing an analyte-containing sample into the microchamber device of the analysis device, irradiating the sample with excitation light corresponding to the compound contained in the sample from a light source, and detecting the emission originating from the compound excited by the excitation light by imaging the microchamber device with the imaging device.

[0186] The substance to be analyzed by the analytical device of this embodiment is not particularly limited, but may be, for example, nucleic acids, proteins, sugars, lipids and complexes thereof, as well as viruses. Alternatively, it may be a substance that can be labeled with a fluorescent dye or quantum dots. Examples of fluorescent dyes include fluorescent nucleic acid aptamers and fluorescent proteins. The target substance is preferably nucleic acid, protein, sugar, lipid and complexes thereof, which can serve as markers for various diseases or infectious diseases. Nucleic acids include natural nucleic acids such as DNA and RNA, and artificial nucleic acids such as LNA and PNA, and polymers thereof are also included. Nucleic acids can be labeled with fluorescent nucleic acid aptamers and detected optically. In a measurement system where the concentration of analyte is low to perform quantitative analysis at the single-molecule level, when the substance to be analyzed is nucleic acid, the intensity of luminescence derived from the compound excited by the excitation light tends to be weaker. However, since the analytical method of this embodiment has high measurement sensitivity, a highly accurate digital assay can be realized even when the substance to be analyzed is nucleic acid in the analytical method of this embodiment. Similarly, when the substance to be analyzed is labeled with one or more elements selected from the group consisting of fluorescent nucleic acid aptamers and fluorescent proteins, the intensity of the luminescence originating from the compound excited by the excitation light tends to be particularly weak. However, in the analytical method of this embodiment, a highly accurate digital assay can be achieved even when measuring such substances.

[0187] (Introduction of analyte-containing samples) In the analysis method of this embodiment, first, the imaging device is optionally placed in the imaging device holder 440, and then the analyte-containing sample is introduced into the microchamber device 300. This introduction may be performed before or after placing the microchamber device 300 in the device holder 320. For example, after placing an appropriate amount of the analyte-containing sample into the microchamber device 300, the analyte-containing sample can be introduced by closing the opening of the microchamber device 300 with the lid.

[0188] If the microchamber device 300 includes a bottom portion having a plate-like member and side walls having a hydrophobic upper surface, and a lid portion provided opposite the bottom portion, and a space consisting of a channel and a well is formed within the microchamber device 300, then the analyte-containing sample can be introduced into the microchamber device 300 as follows. The following explanation will be given with reference to Figure 9 as appropriate.

[0189] First, sample 308 containing analyte 308a is introduced into the microchamber device 300. Figure 9 shows an example in which chromogenic substrate 308b, which reacts with or binds to analyte 308a to produce fluorescence, is introduced.

[0190] Sample 308 contains a suitable solvent for dissolving or suspending the analyte and chromogenic substrate, and solvents commonly used for detecting nucleic acids, proteins, sugars, lipids and their complexes, as well as viruses, can be used. Examples of such solvents include water, alcohol, ether, ketone, nitrile solvents, dimethyl sulfoxide (DMSO), and N,N-dimethylformamide (DMF), with water being preferred.

[0191] The sample 308, containing the analyte and chromogenic substrate, can be injected, for example, through an inlet provided on the lid 310 and connected to the inside of the microchamber device 300. An outlet for discharging the solvent and gas may be connected to the opposite side of the inlet. The sample 308 introduced into the microchamber device 300 travels through the space inside the microchamber device 300 and fills the microchamber device 300. Thus, the sample 308 containing the analyte 308a and chromogenic substrate 308b is introduced into the channel 312 and each well 302.

[0192] If the concentration of analyte in sample 308 is low, some wells will contain only one molecule of analyte, while others will contain none. Conversely, if the concentration of analyte in sample 308 is higher, each well may contain two or more molecules of analyte. On the other hand, it is preferable that the chromogenic substrate is included in sample 308 at a concentration sufficiently higher than that of the analyte. In this case, the chromogenic substrate is introduced into almost all of the wells.

[0193] Next, a hydrophobic solution or gas is introduced into the microchamber device 300. The hydrophobic solution or gas is injected through the same or a different inlet as sample 308 and discharged through the same or a different outlet as sample 308. The hydrophobic solution or gas introduced into the microchamber device 300 proceeds through the microchamber device 300, displacing the sample 308 that was filling the flow path of the microchamber device 300, but without displacing the sample 308 in the wells. As a result, droplets of sample 308 of a defined volume are formed in the wells. By adjusting the concentration of analyte in the sample 308, some of the droplets formed in each well 302 contain analyte 308a along with the chromogenic substrate 308b.

[0194] (Irradiation with excitation light) Next, excitation light corresponding to the compounds contained in the analyte-containing sample in the microchamber device 300 is irradiated from the light source 500. For example, if the analyte-containing sample contains a fluorescent substance, light having a wavelength corresponding to the excitation light of that fluorescent substance is irradiated from the light source 500 onto the microchamber device 300.

[0195] In the introduction of an analyte-containing sample, when the sample containing the analyte and chromogenic substrate is introduced into a well provided on the first surface of the microchamber device 300, excitation light corresponding to the compound produced by the reaction between the analyte and the chromogenic substrate may be irradiated. For example, the analyte may be a substance having an enzyme on its surface or internally that has substrate cleavage activity for the chromogenic substrate, and the chromogenic substrate may be a substance that is cleaved by the enzyme to release a reaction product as a chromophore. Alternatively, the analyte and the chromogenic substrate may form a complex to produce a reaction product as a chromophore. The chromogenic substrate may be, for example, a substance that produces a reaction product having different optical properties after reaction with an enzyme, a substance whose light scattering intensity or optical rotation changes before and after the reaction, or a substance that exhibits fluorescence to excitation light of a specific wavelength after the reaction. The chromogenic substrate does not have to be a single substance, and may be, for example, a mixture of an enzyme that replicates and amplifies the analyte, a replication substrate, and a fluorescent dye that binds to the amplified analyte and fluoresces.

[0196] To promote the reaction between the analyte and the chromogenic substrate, the microchamber device 300 into which the sample is introduced may be subjected to a temperature cycle treatment of heating, cooling, or repeated heating and cooling. These temperature treatments may be performed before setting the microchamber device 300 inside the analytical device, or they may be performed inside the analytical device after setting the microchamber device 300 inside the analytical device.

[0197] For example, if the analyte is a nucleic acid, the chromogenic substrate may be a fluorescent dye for nucleic acids. Examples of fluorescent dyes for nucleic acids include Ethidium Bromide, Acridine Orange, SYBR Green, EvaGreen, TO1-Biotin, DFHBI, DFHBI-1T, SHERLOCK probe, and DETECTR probe.

[0198] (Imaging of microchamber devices) Next, the imaging device 400 images the microchamber device 300 to detect luminescence derived from the compound excited by the irradiation of the excitation light described above.

[0199] In the introduction of the analyte-containing sample, when a sample containing an analyte and a chromogenic substrate is introduced into the wells provided on the first surface of the microchamber device 300, in the droplets formed in each well, the reaction between the analyte and the chromogenic substrate proceeds and reaction products are generated only in the wells where both coexist. Here, when the volume of the well is extremely small, for example, on the order of 10 aL to 100 nL, the concentration of the trace amount of reaction products generated from the reaction of one analyte molecule with the chromogenic substrate becomes high. Therefore, when the excitation light is irradiated, high-intensity luminescence that can be detected by the imaging device 400 is generated.

[0200] Therefore, in the introduction of the analyte-containing sample, when a sample containing an analyte and a chromogenic substrate is introduced into the wells provided on the first surface of the microchamber device 300, quantitative analysis at the single-molecule level can be performed.

[0201] (Analysis of imaging data) The imaging data obtained as described above may be analyzed by an information processing device connected to or integrated with the imaging device 400. For example, the concentration of the analyte in the sample in the microchamber device 300 may be analyzed by measuring the luminance or brightness of the imaging data.

[0202] In the introduction of the analyte-containing sample, when a sample containing an analyte and a chromogenic substrate is introduced into the wells provided on the first surface of the microchamber device 300, the concentration of the analyte in the sample may be analyzed by measuring the number of wells that emit light in response to the excitation light among the plurality of imaged wells. Also, not only by measuring the number of wells that emit light, but by measuring the luminance or brightness of each well, it may be analyzed how many molecules of the analyte have been introduced into each well.

[0203] [Numerical Examples 1 to 5] The numerical examples of the aspherical lens 100 described above will be described below, but the present invention is not limited thereto. In the following numerical examples, the unit of length is shown in mm.

[0204] Each parameter of the optical system is shown in FIG. 10. R1 to R10 are, in order, the first main surface of the microchamber device 300, the second main surface of the microchamber device 300, the first main surface of the first optical filter 200, the second main surface of the first optical filter 200, the first main surface of the first lens 101, the second main surface of the first lens 101, the first main surface of the second lens 102, the second main surface of the second lens 102, the first main surface of the third lens 103, and the second main surface of the third lens 103. Also, R11 means the arrangement position of the imaging lens. Here, a plurality of wells are arranged on the surface R1.

[0205] Also, in Numerical Examples 1 to 5, the refractive index and Abbe number of the microchamber device 300 are 1.531 and 56.0, respectively, the refractive index and Abbe number of the first optical filter 200 are 1.523 and 54.5, respectively, and the refractive index and Abbe number of the first to third lenses are the values described in Table 8 below. Note that the materials used for the microchamber device, the first optical filter, and each lens are not particularly limited as long as they satisfy the physical property values described in Numerical Examples 1 to 5. For example, the refractive index and Abbe number of the microchamber device 300, and the first lens 101 and the third lens 103 can be obtained by a cycloolefin polymer (COP) or the like. Examples of such first lens 101 and third lens 103 include ZEONEX manufactured by Nippon Zeon Co., Ltd. The refractive index and Abbe number of the first optical filter 200 can be obtained by a general commercially available UV cut filter or the like. The refractive index and Abbe number of the second lens 102 can be obtained by a commercially available lens or the like. Examples of such second lens 102 include Yupizer manufactured by Mitsubishi Gas Chemical Company, Inc. Note that at least Numerical Examples 1 and 3 are numerical examples when all lenses are made of non-fluorescent materials.

[0206] The distances d1 to d10 in Numerical Examples 1 to 5 are shown in the following table. [Table 1]

[0207] In numerical examples 1 to 5, R1, R2, R3, and R4 were assumed to be planes. R5, R6, R7, R8, R9, and R10 were aspherical surfaces, and each surface is represented by the following aspherical function. In the following formulas, the direction of the optical axis is taken as z, the direction perpendicular to the optical axis is taken as y, K is the conicity coefficient, R is the radius of curvature, and a04, a06, a08, a10, a12, a14, and a16 are the aspherical coefficients.

number

[0208] In numerical examples 1 to 5, the conicity coefficient K in the above formula was set to 0, and the radii of curvature R of surfaces R5 to R10 were set to the values ​​shown in the table below. Note that a negative radius of curvature means that the curvature is convex to the right in Figure 10. [Table 2]

[0209] Furthermore, the aspherical coefficients a04, a06, a08, a10, a12, a14, and a16 in numerical examples 1 to 5 were set to the values ​​shown in the table below. In the table below, "E±x" means "×10± x It means "...".

[0210] (Numerical Example 1) [Table 3]

[0211] (Numerical Example 2) [Table 4]

[0212] (Numerical Example 3) [Table 5]

[0213] (Numerical Example 4) [Table 6]

[0214] (Numerical Example 5) [Table 7]

[0215] The parameters in Numerical Examples 1 to 5 were the values shown in the following table. The definitions of the symbols in the following table are as described above. [Table 8]

[0216] Also, as the overall characteristics of the aspherical lens composed of the first lens 101, the second lens 102, and the third lens 103 in each numerical example, the focal length f, the effective F-number, and the half angle ω were the values shown in the following table. From the above, it was found that in Numerical Examples 1 to 5, even when mounted on an existing imaging lens, the overall optical performance does not deteriorate, and it is possible to observe a fine object with high resolution. [Table 9]

[0217] [Numerical Examples 6 to 11] Schematic cross-sectional views of the optical systems corresponding to Numerical Examples 6 to 11 below are shown in FIGS. 11 to 16. The parameters of the optical systems are as shown in FIG. 10 in the same manner as in Numerical Examples 1 to 5 above.

[0218] In numerical examples 6 to 11, the refractive index and Abbe number of the microchamber device 300 were set to 1.531 and 56.0, respectively, the refractive index and Abbe number of the first optical filter 200 were set to 1.523 and 54.5, respectively, and the refractive index and Abbe numbers of the first to third lenses were the values ​​shown in Table 18 below. The materials used for the microchamber device, the first optical filter, and each lens are not particularly limited as long as they satisfy the physical properties described in Numerical Examples 6 to 11. For example, a refractive index of 1.531 and an Abbe number of 56.0, as in the microchamber device 300, can be obtained from cycloolefin polymer (COP), etc. An example of a COP lens is ZEONEX manufactured by Nippon Zeon Co., Ltd. The refractive index and Abbe number of the first optical filter 200 can be obtained from a general commercially available UV-cut filter, etc. Lenses with other refractive indices and Abbe numbers can be obtained from commercially available lenses, etc. Examples of such lenses include Yupizeta manufactured by Mitsubishi Gas Chemical Co., Ltd., OKP manufactured by Osaka Gas Chemical Co., Ltd., APL manufactured by Mitsui Chemicals, and glass lenses (manufactured by Ohara, SCHOTT, HOYA, HIKARI, etc.). Numerical Examples 6 to 11 are numerical examples in which all lenses are made of non-fluorescent materials.

[0219] The distances d1 to d10 in numerical examples 6 to 11 are shown in the table below. [Table 10]

[0220] In numerical examples 6 to 11, R1, R2, R3, and R4 were assumed to be planes. R5, R6, R7, R8, R9, and R10 were aspherical surfaces, and each surface is represented by the following aspherical function. In the following formulas, the direction of the optical axis is taken as z, the direction perpendicular to the optical axis is taken as y, K is the conicity coefficient, R is the radius of curvature, and a04, a06, a08, a10, a12, a14, and a16 are the aspherical coefficients.

number

[0221] In numerical examples 6 to 11, the radii of curvature R of surfaces R5 to R10 were set to the values ​​shown in the table below. Note that a negative radius of curvature means that the curvature is convex to the right in Figure 10. [Table 11]

[0222] Furthermore, the conicity coefficient K and asphericity coefficients a04, a06, a08, a10, a12, a14, and a16 in numerical examples 6 to 11 were the values ​​shown in the table below. In the table below, "E±x" means "×10± x It means "...".

[0223] (Numerical Example 6) [Table 12]

[0224] (Example 7 of numerical values) [Table 13]

[0225] (Numerical Example 8) [Table 14]

[0226] (Numerical Example 9) [Table 15]

[0227] (Numerical Example 10) [Table 16]

[0228] (Numerical Example 11) [Table 17]

[0229] The parameters in numerical examples 6 to 11 were as shown in the table below. The definitions of the symbols in the table below are as described above. [Table 18]

[0230] Furthermore, the overall characteristics of the aspherical lens consisting of the first lens 101, the second lens 102, and the third lens 103 in numerical examples 6 to 11 were as shown in the table below: focal length f, effective F-number, and half-angle of view ω. From the above, it was found that in numerical examples 6 to 11, even when the object to be photographed is at close range, a wide area can be observed even when attached to an existing imaging lens, and the overall optical performance does not deteriorate, making it possible to observe minute objects with high resolution. [Table 19]

[0231] [Note] The present invention includes the following embodiments. [1] A device holder for holding a microchamber device, A light source that irradiates the first surface of the microchamber device with laser light from diagonally above or diagonally below, One or more aspherical lenses are arranged such that their optical axes are substantially parallel to the normal of the first surface of the microchamber device, An imaging device for imaging the microchamber device through the aspherical lens, A first optical filter located between the microchamber device and the imaging device, the first optical filter blocking the laser light emitted by the light source and transmitting at least a portion of the light with wavelengths longer than the laser light emitted by the light source, An analytical device equipped with the following features. [2] The system includes a focusing lens that concentrates the laser light emitted from the aforementioned light source, The laser light irradiated onto the microchamber device is laser light irradiated from the light source and focused after passing through the focusing lens. [1] The analytical device described above. [3] The angle between the first surface of the microchamber device and the central axis of the laser beam irradiated onto the microchamber device is 3 to 40°. The analytical device described in [1] or [2]. [4] An analytical device according to any one of [1] to [3], comprising a plurality of the aforementioned aspherical lenses. [5] The plurality of aspherical lenses are, from the microchamber device side, a first lens having positive refractive power, a second lens having negative refractive power, and a third lens having positive refractive power. Both sides of the first lens, the second lens, and the third lens are aspherical, Multiple aspherical lenses satisfy the following formulas (1), (2), (3), (4), (5), and (6): [4] The analytical device described above. (1) 4.30 ≤ f1 / f ≤ 4.95 (2) 0.120 ≤ φ1 / n1 ≤ 0.160 (3) 0.0030 ≤ φ1 / ν1 ≤ 0.0050 (4) -3.20 ≤ f² / f ≤ -2.70 (5) -0.240 ≤ φ2 / n2 ≤ -0.160 (6) -0.0200 ≤ φ2 / ν2 ≤ -0.0120 f: Total focal length of the attachment lens relative to the d line f1: Focal length of the first lens relative to the d line φ1: The product of the power of the first lens with respect to the d line and f (f / f1) n1: Refractive index of the first lens with respect to the d line ν1: Abbe number of the first lens f2: Focal length of the second lens relative to the d line φ2: The product of the power of the second lens with respect to the d line and f (f / f2) n2: Refractive index of the second lens with respect to the d line ν2: Abbe number of the second lens [6] The plurality of aspherical lenses are, from the microchamber device side, a first lens having a positive or negative refractive power, a second lens having a positive refractive power, and a third lens having a positive or negative refractive power. Both sides of the first lens, the second lens, and the third lens are aspherical, Multiple aspherical lenses satisfy the following formulas (I), (II), (III), (IV), (V), and (VI): [4] The analytical device described above. (I)-10.0≦f1 / f≦8.00 (II)-0.1500≦φ1 / ν1≦0.0500 (III) 0.100 ≤ φ2 / n2 ≤ 2.000 (IV) -80.0 ≤ f3 / f ≤ 30.0 (V)-0.300≦φ3 / n3≦0.900 (VI)-0.0200≦φ3 / ν3≦0.0500 f: Total focal length of the attachment lens relative to the d line f1: Focal length of the first lens relative to the d line φ1: The product of the power of the first lens with respect to the d line and f (f / f1) ν1: Abbe number of the first lens f2: Focal length of the second lens relative to the d line φ2: The product of the power of the second lens with respect to the d line and f (f / f2) n2: Refractive index of the second lens with respect to the d line f3: Focal length of the third lens relative to the d line φ3: The product of the power of the third lens on the d line and f (f / f3) n3: Refractive index of the third lens with respect to the d line ν3: Abbe number of the third lens [7] The analytical device according to any one of [1] to [6], wherein the aspherical lens is made of a non-fluorescent material. [8] The analytical device according to any one of [1] to [7], wherein the device holder or the aspherical lens is movable in the direction normal to the first surface of the microchamber device. [9] The analytical device according to any one of [1] to [8], wherein a plurality of wells are provided on the first surface of the microchamber device.

[10] The imaging device further comprises an information processing device connected to or integrated with the imaging device, The analysis device according to [9], wherein the information processing device detects a well among the plurality of imaged wells that emits light in response to the laser light.

[11] Introducing an analyte-containing sample into the microchamber device of the analytical device described in any of [1] to

[10] , The process involves irradiating the sample with excitation light corresponding to the compound contained in the sample from the light source, By imaging the microchamber device with the imaging device, the light emission originating from the compound excited by the excitation light is detected. A method for analyzing a sample, including [specific details].

[12] The aforementioned analyte-containing sample contains a fluorescent dye or quantum dots.

[11] by the method described in

[11] .

[13] The method according to

[11] or

[12] , wherein the first optical filter blocks light having a wavelength corresponding to the excitation light of the compound and transmits light having a wavelength corresponding to the light emitted by the excited compound.

[14] A device holder for holding a microchamber device, A light source that irradiates the first surface of the microchamber device with laser light from diagonally above or diagonally below, One or more aspherical lenses are arranged such that their optical axes are substantially parallel to the normal of the first surface of the microchamber device, An imaging device holding unit for holding the imaging device in a position where the microchamber device can be imaged via the aspherical lens, A first optical filter located between the microchamber device and the held imaging device, the first optical filter blocking the laser light emitted by the light source and transmitting at least a portion of the light on the longer wavelength side than the laser light emitted by the light source, An analytical device equipped with the following features. [Examples]

[0232] The present invention will be described more specifically below using examples and comparative examples. The present invention is not limited in any way by the following examples.

[0233] [Example 1] An analytical device with the configuration shown in Figure 8 was fabricated. First, the observation optical system was fabricated. The main rear camera of a smartphone (iPhone® 15 Pro) was used as the imaging device. The smartphone was also used as an information processing device. The imaging conditions were ISO sensitivity 1250, exposure 1.0 second, and spot metering. The objective lens used as the aspherical lens 110 on the microchamber device side was a THORLABS C0390TME-A, and the eyepiece lens used as the aspherical lens 100 on the imaging device side was a THORLABS C340TMD-A. A first optical filter (cut wavelength 510 nm, made by stacking two short-wavelength cut filters manufactured by Asahi Spectroscopic Co., Ltd.) was placed between the eyepiece lens and the objective lens. The smartphone was positioned so that the lens of the main rear camera was directly above the eyepiece lens. A TYLASER 488 nm (50 mW) laser device was used as the light source. The laser device was positioned diagonally above the first surface (observation surface) of the microchamber device, as described later, such that the angle A between the first surface of the microchamber device and the central axis of the laser beam irradiated onto the microchamber device is 4°. In this way, the observation optical system was fabricated.

[0234] The angle A was adjusted as follows: First, it was confirmed that the central axis of the laser beam emitted from the light source placed on a flat surface was parallel to that surface. Then, the angle A was adjusted by changing the angle between the surface on which the light source was placed and the first surface using a THORLABS GN05 / M goniometric stage. For example, when the angle between the surface on which the light source was placed and the first surface was set to 4°, the angle A also became 4°.

[0235] The microchamber device was fabricated as follows: First, a fluororesin film (Asahi Glass Co., Ltd., CYTOP, Cat No.: CTL-816AP) was spin-coated onto a glass substrate (Matsunami Glass Co., Ltd., cover glass, 24 mm x 32 mm, thickness 0.17 mm, refractive index 1.5). The spin-coating conditions were 2000 rpm for 30 seconds. A glass substrate with a fluororesin coating was baked at 180°C for 1 hour to ensure close adhesion between the fluororesin coating and the glass substrate. The spin coating and baking process was repeated a total of four times to form a fluororesin coating approximately 3 μm thick on the glass substrate. A photoresist film was formed by spin-coating a positive-type photoresist (AZ Electronic Materials, AZP4903) onto a fluororesin film. The spin-coating conditions were 4000 rpm for 60 seconds. A glass substrate with a photoresist film formed on it was baked at 110°C for 1 hour to ensure close adhesion between the photoresist film and the fluororesin film.

[0236] Next, a chamber shape pattern was formed using photolithography. The photoresist film was irradiated with ultraviolet light through a mask patterned with the chamber shape. The glass substrate was immersed in a resist developer solution to remove the portion of the photoresist film that had been irradiated with ultraviolet light. The substrate was placed in a dry etching apparatus (SAMCO Corporation, RIE-10NR), and dry etching was performed using O2 plasma to remove the fluororesin coating in the resist removal area, exposing the glass on the bottom surface. The glass substrate was cleaned with acetone and ethanol to completely remove the photoresist film, and a chamber array chip with a patterned chamber shape was obtained. The resulting chamber array chip has approximately 5.7 million chambers, each with a diameter of 4 μm and a height of 3 μm, formed in a 20 x 20 mm chamber array area at a repeating pitch of 9 μm.

[0237] A microchamber device was fabricated by attaching double-sided tape cut to the shape of the channel onto a chamber array chip, and then bonding a 5mm thick perforated glass that had been spin-coated with fluororesin (Asahi Glass Co., Ltd., CYTOP, Cat No.: CTL-809M). The two holes in the perforated glass serve as the inlet and outlet.

[0238] In the microchamber device obtained as described above, fluorescent beads (Fluoresbrite YG 1.0 μm, Polysciences) were dispersed in a buffer solution and then flowed in. After that, laser light was irradiated, and observation was performed using the observation optical system described above.

[0239] [Examples 2-10] The observation was carried out in the same manner as in Example 1, except that the angle A between the first surface and the central axis of the laser beam irradiated onto the microchamber device was adjusted to the values ​​shown in the table below.

[0240] [Table 20]

[0241] [Comparative Example 1] The observation was carried out in the same manner as in Example 1, except that the angle A between the first surface and the central axis of the laser beam irradiated onto the microchamber device was adjusted to be 0°. In this case, the analyte is irradiated with excitation light by evanescent waves generated by light that has been totally reflected and guided within the microchamber device.

[0242] [Example 11] An analytical device with the configuration shown in Figure 7 was fabricated. First, the observation optical system was constructed. The main rear camera of a smartphone (iPhone® 15 Pro) was used as the imaging device. The smartphone was also used as an information processing device. The imaging conditions were ISO sensitivity 1250, exposure 1.0 second, and spot metering. A THORLABS C060TMD-A was used as the focusing lens, a THORLABS C0390TME-A was used as the objective lens as an aspherical lens on the microchamber device side, and a THORLABS C340TMD-A was used as the eyepiece lens as an aspherical lens on the imaging device side. A first optical filter (cut wavelength 510 nm, made by stacking two short-wavelength cut filters manufactured by Asahi Spectroscopic Co., Ltd.) was placed between the eyepiece lens and the objective lens. The smartphone was positioned so that the lens of the main rear camera was directly above the eyepiece lens. A TYLASER 488 nm (50 mW) laser device was used as the light source. The focusing lens and laser device were positioned so that the angle between the central axis of the laser beam incident on the focusing lens and the horizontal plane was 3°. Using the distance between the rotation axis of the laser device and the focusing lens (32 mm), the effective focal length of the focusing lens (9.6 mm), and the distance between the observation position in the microchamber device (i.e., the laser incident position on the first surface) and the focusing lens (approximately 14 mm), the angle A between the first surface of the microchamber device and the central axis of the laser beam irradiated onto the microchamber device was calculated to be approximately 7°. The observation optical system was fabricated in this manner.

[0243] The observation was performed in the same manner as in Example 1, except that the observation optical system was as described above.

[0244] [Examples 12-13] Except for adjusting the angle of the laser device so that the angle between the central axis of the laser beam incident on the focusing lens and the horizontal plane is 4° (Example 12) or 5° (Example 13), observations were performed in the same manner as in Example 11. The angle A between the first plane and the central axis of the laser beam irradiated onto the microchamber device is calculated in the same manner as in Example 11, resulting in approximately 9° (Example 12) and approximately 12° (Example 13).

[0245] The observation results are shown in Figure 17. As shown in Figure 17, the observation images in the example were observed with higher measurement sensitivity compared to the observation images in the comparative example.

[0246] [Example 14] The same observation optical system as in Example 12 was used, except that the rear main camera of the iPhone® 8 was used instead of the rear main camera of the iPhone® 15 Pro. The imaging conditions were ISO sensitivity 4000, exposure 1.0 second, and spot metering. Furthermore, nucleic acid detection was performed using the same microchamber device as in Example 12, as described below. The R-phycoerythrin beads (R-PE beads) used for alignment were fabricated by biotinizing polystyrene beads (made by micromod) with amino groups on their surface and a diameter of 3 μm using EZ-Link NHS-PEG4-Biotin (Thermo Scientific), and then attaching Streptavidin R-PE Conjugate (made by TCI). As the analyte (target) for detection, a DNA fragment (181 bp) obtained by amplified by a PCR reaction of a portion of the N gene of the SARS-CoV-2 virus was used. As the chromogenic reagent, we utilized a system in which the target DNA was amplified by a Recombinase Polymerase Amplification reaction using a forward primer containing the T7 promoter sequence and a reverse primer containing the Mango Aptamer sequence. Then, an RNA molecule containing the Mango Aptamer sequence was generated from the amplified DNA using T7 polymerase, and this molecule fluoresces when it binds to the chromogenic substrate TO1-Biotin. The nucleic acid detection reaction sample solutions were prepared as follows: Reaction premix A consisted of 2.1 μL of a forward primer with a T7 promoter sequence at a concentration of 10 μM, 2.1 μL of a reverse primer with a Mango Aptamer sequence at a concentration of 10 μM, one TwistAmp Basic pellet (TwistDX), 29.5 μL of TwistAmp Rehydration Buffer (TwistDX), 0.75 μL of TT7 polymerase (1000 U / μL, TOYOBO), 1 μL of TO-1 Biotin (50 μM, abm), 2 μL of NTP (25 mM, Invitrogen), and 0.5 μL of S-386 (1% (v / v), AGC Seimi Chemical). Water was added to bring the volume to 42.5 μL. Reaction premix B was a 280 mM magnesium acetate solution (TwistDX). 17 μL of reaction premix A, 1 μL of analyte solution (final concentration 300 fM), 1 μL of R-PE bead solution, and 1 μL of reaction premix B were mixed to prepare the reaction sample solution. The reaction sample solution was immediately injected into the microchamber device. Next, a hydrophobic liquid (Fomblin YLVAC 06 / 6, manufactured by Solvay, with 0.1% S-386 (v / v) added) was injected into the microchamber device to isolate the microchamber and separate the reaction sample solution. After that, the microchamber device was held at 39°C for 30 minutes to promote the nucleic acid detection reaction, and then laser light was irradiated and observation was performed using the observation optical system described above. Focusing was performed by manually fine-tuning the installation height of the microchamber device while observing the fluorescence of the R-PE beads, which exhibit bright yellow fluorescence, with the imaging device.

[0247] [Comparative Example 2] The observation was performed in the same manner as in Example 14, except that the angle A between the first surface and the central axis of the laser beam irradiated onto the microchamber device was adjusted to be 0°. The imaging conditions were ISO sensitivity 2000, exposure 1.0 second, and spot metering.

[0248] The observation results are shown in Figure 18. As shown in Figure 18, nucleic acids were observed with high measurement sensitivity in Example 14. In Comparative Example 2, what was observed was R-PE beads used for alignment and was not derived from nucleic acids. [Explanation of Symbols]

[0249] 10, 20, 30, 40, 50, 60, 70…Analysis device, 100…Aspherical lens, 110…Lens, 101…First lens, 102…Second lens, 103…Third lens, 200…First optical filter, 300…Microchamber device, 302…Well, 304…Side wall, 304a…Top surface, 306…Plate-shaped member, 308…Sample, 308a…Analyte, 308b…Chromogenic substrate, 310…Lid, 312…Flow channel, 320…Device holder, 400…Imaging device, 420…Imaging lens, 440…Imaging device holder, 460…Hole, 500…Light source, 510…Concentrating lens, 520…Waveguide, 600, 610…Light shielding means.

Claims

1. A device holder for holding a microchamber device, A light source that irradiates the first surface of the microchamber device with laser light from diagonally above or diagonally below, One or more aspherical lenses are arranged such that their optical axes are substantially parallel to the normal of the first surface of the microchamber device, An imaging device for imaging the microchamber device through the aspherical lens, A first optical filter located between the microchamber device and the imaging device, the first optical filter blocking the laser light emitted by the light source and transmitting at least a portion of the light on the longer wavelength side than the laser light emitted by the light source, An analytical device equipped with the following features.

2. The system includes a focusing lens that concentrates the laser light emitted from the aforementioned light source, The laser light irradiated onto the microchamber device is laser light irradiated from the light source and focused after passing through the focusing lens. The analytical device according to claim 1.

3. The angle between the first surface of the microchamber device and the central axis of the laser beam irradiated onto the microchamber device is 3 to 40°. The analytical device according to claim 1.

4. The analytical device according to claim 1, comprising a plurality of aspherical lenses.

5. The plurality of aspherical lenses, from the microchamber device side, consist of a first lens having positive refractive power, a second lens having negative refractive power, and a third lens having positive refractive power. Both sides of the first lens, the second lens, and the third lens are aspherical, Multiple aspherical lenses satisfy the following formulas (1), (2), (3), (4), (5), and (6): The analytical device according to claim 4. (1) 4.30 ≤ f1 / f ≤ 4.95 (2) 0.120 ≤ φ1 / n1 ≤ 0.160 (3) 0.0030 ≤ φ1 / ν1 ≤ 0.0050 (4) -3.20 ≤ f² / f ≤ -2.70 (5) -0.240 ≤ φ2 / n2 ≤ -0.160 (6) -0.0200 ≤ φ² / ν² ≤ -0.0120 f: Total focal length of the attachment lens relative to the d line f1: Focal length of the first lens relative to the d line φ1: The product of the power of the first lens with respect to the d line and f (f / f1) n1: Refractive index of the first lens with respect to the d line ν1: Abbe number of the first lens f2: Focal length of the second lens relative to the d line φ2: The product of the power of the second lens with respect to the d line and f (f / f2) n2: Refractive index of the second lens with respect to the d line ν²: Abbe number of the second lens

6. The plurality of aspherical lenses are, from the microchamber device side, a first lens having positive or negative refractive power, a second lens having positive refractive power, and a third lens having positive or negative refractive power. Both sides of the first lens, the second lens, and the third lens are aspherical, Multiple aspherical lenses satisfy the following formulas (I), (II), (III), (IV), (V), and (VI): The analytical device according to claim 4. (I) -10.0≦f1 / f≦8.00 (II) -0.1500≦φ1 / ν1≦0.0500 (III) 0.100≦φ2 / n2≦2.000 (IV) -80.0≦f3 / f≦30.0 (V)-0.300≦φ3 / n3≦0.900 (VI) -0.0200≦φ3 / ν3≦0.0500 f: Total focal length of the attachment lens relative to the d line f1: Focal length of the first lens relative to the d line φ1: The product of the power of the first lens with respect to the d line and f (f / f1) ν1: Abbe number of the first lens f2: Focal length of the second lens relative to the d line φ2: The product of the power of the second lens with respect to the d line and f (f / f2) n2: Refractive index of the second lens with respect to the d line f3: Focal length of the third lens relative to the d line φ3: The product of the power of the third lens with respect to the d line and f (f / f3) n3: Refractive index of the third lens with respect to the d line ν3: Abbe number of the third lens

7. The analytical device according to claim 1, wherein the aspherical lens is made of a non-fluorescent material.

8. The analytical device according to claim 1, wherein the device holder or the aspherical lens is movable in the direction normal to the first surface of the microchamber device.

9. The analytical device according to claim 1, wherein a plurality of wells are provided on the first surface of the microchamber device.

10. The imaging device further comprises an information processing device connected to or integrated with the imaging device, The analysis device according to claim 9, wherein the information processing device detects a well among the plurality of imaged wells that emits light in response to the laser light.

11. Introducing an analyte-containing sample into the microchamber device of the analytical device according to any one of claims 1 to 10, The process involves irradiating the sample with excitation light corresponding to the compound contained in the sample from the light source, By imaging the microchamber device with the imaging device, the light emission originating from the compound excited by the excitation light is detected. A method for analyzing a sample, including [specific details].

12. The aforementioned analyte-containing sample contains a fluorescent dye or quantum dots. The method according to claim 11.

13. The method according to claim 11, wherein the first optical filter blocks light having a wavelength corresponding to the excitation light of the compound and transmits light having a wavelength corresponding to the light emitted by the excited compound.

14. A device holder for holding a microchamber device, A light source that irradiates the first surface of the microchamber device with laser light from diagonally above or diagonally below, One or more aspherical lenses are arranged such that their optical axes are substantially parallel to the normal of the first surface of the microchamber device, An imaging device holding unit for holding the imaging device in a position where the microchamber device can be imaged via the aspherical lens, A first optical filter located between the microchamber device and the held imaging device, the first optical filter blocking the laser light emitted by the light source and transmitting at least a portion of the light on the longer wavelength side than the laser light emitted by the light source, An analytical device equipped with the following features.

Images