JP6867320B2 - Light measuring device and light measuring method - Google Patents

Description

The present invention relates to an optical plate, a light irradiation device, a light measurement device, a light irradiation method, and a light measurement method.

Patent Document 1 describes a light irradiation device that irradiates a microplate holding a measurement object in a plurality of wells with light. This light irradiation device includes a light guide member having a main surface on which a plurality of convex portions are formed, a back surface which is a surface opposite to the main surface, and a side surface substantially orthogonal to the main surface. Further, this light irradiation device includes a light source that injects light into the light guide member from the side surface of the light guide member. The light guide member is arranged so that the upper surface of the convex portion is in contact with the back surface of the microplate.

JP-A-2010-230397

In the light irradiation device described in Patent Document 1, the light incident on the light guide member from the side surface of the light guide member is reflected by the main surface and the back surface of the light guide member and is incident on the convex portion, and then the convex portion. It is incident on the well of the macro plate from the upper surface. At this time, light inclined with respect to the depth direction of the well is incident on the well of the microplate. Therefore, it is possible to uniformly irradiate the object to be measured with the light as compared with the case where the light is incident along the depth direction of the well.

By the way, in order to uniformly irradiate the object to be measured with light using the above-mentioned light guide member, it is necessary to process the shape of the convex portion according to the arrangement and shape of the well holding the object to be measured. There is. However, in reality, when the light guide member is made of quartz or the like, a relatively high processing technique is required for processing to form the convex portion, and it is difficult to process it into a desired shape. In other words, it is not easy to process the convex portion so as to irradiate light according to a desired irradiation condition.

The present invention has been made in view of such circumstances, and is an optical plate capable of irradiating light according to a desired irradiation condition, a light irradiation device, a light measurement device, a light irradiation method, and light. It is an object of the present invention to provide a measuring method.

In order to solve the above problems, the optical plate according to the present invention is an optical plate for irradiating an object with light, and has an optical input surface for inputting light (input light) and light (output light). ), A back surface facing the light output surface, and a light diffusing portion formed at least inside the optical plate by condensing laser light to diffuse the light. The optical input surface is a surface between the optical output surface and the back surface, and the light input from the optical input surface (input light) is diffused in the light diffusing portion and output from the optical output surface. And.

This optical plate includes a light diffusing portion that diffuses the light input from the light input surface and outputs the light from the light output surface. That is, the light (input light) input from the optical input surface is diffused inside the optical plate in the light diffusing portion, and is output from the optical output surface as output light. Therefore, if the optical plate is arranged so that the light output surface faces the object, the object can be irradiated with the output light from the light output surface. In particular, in this optical plate, a light diffusing portion is formed by condensing laser light on a member serving as an optical plate (that is, by laser processing). Therefore, it is possible to irradiate the member with light according to a desired irradiation condition, as compared with the case where a convex portion or a concave portion is actually formed on the member.

The optical plate according to the present invention may have a light guide portion that guides light between an optical input surface and a surface facing the optical input surface. In this case, the light input from the light input surface is diffused and output by the light diffusing unit while being guided between the light input surface and the opposite surface by the light guide unit. Therefore, it is possible to more uniformly irradiate an object existing in various places on the region where the optical plate is arranged.

The optical plate according to the present invention includes a first portion in which a light diffusing portion is formed and a second portion integrally formed with the first portion, and the light guide portion is a second portion. It may be configured in. In this case, if the laser processing is controlled so that the light diffusing portion is not formed in, for example, the second portion of the first and second portions integrally configured with each other, the light guide portion is formed by the second portion. Can be configured. That is, in this case, the light guide portion can be configured in the optical plate by controlling the laser processing.

In the optical plate according to the present invention, the light guide portion is formed by joining a first portion in which a light diffusing portion is formed and a second portion formed separately from the first portion. May be configured in the second part. In this case, if a first portion in which the light diffusing portion is formed by laser processing and, for example, a second portion in which the light diffusing portion is not formed are prepared and joined to each other, the second portion guides the light guide. It can form a part.

In the optical plate according to the present invention, the light diffusing portion may be formed over at least a part between the light output surface and the back surface. In this case, the amount of light output from the region close to the light diffusing portion can be increased.

In the optical plate according to the present invention, the light diffusing portion may be composed of a plurality of light diffusing layers formed by condensing laser light. In this case, the amount of light diffused by the light diffusing portion can be increased.

The optical plate according to the present invention may further include a reflecting portion that extends along a direction intersecting the light output surface and reflects light. In this case, it is possible to irradiate the object with light more uniformly.

The light irradiation device according to the present invention is characterized by including the above-mentioned optical plate and a light source that outputs light (input light) input to the light input surface. Further, the light measuring device according to the present invention measures the light from the above-mentioned light irradiation device, the holding member for holding the object, and the object irradiated with the light (output light) output from the light output surface. It is characterized by including an imaging device for imaging.

Further, the light irradiation method according to the present invention is a surface between the light output surface for outputting light (output light), the back surface facing the light output surface, and the light output surface and the back surface, and is light (input). A light irradiation method for irradiating an object with light using an optical plate including a light input surface on which light) is input and a light diffusing portion formed by condensing laser light, which is applied to the light input surface. By inputting light (input light), the light diffuser diffuses the light and outputs light (output surface) from the light output surface to irradiate the object with light.

Further, the optical measurement method according to the present invention is a surface between an optical output surface for outputting light (output light), a back surface facing the optical output surface, and an optical output surface and the back surface, and is light (input). An object irradiated with light (output light) by irradiating an object with light using an optical plate including an optical input surface on which light) is input and a light diffusing portion formed by condensing laser light. It is an optical measurement method that detects the measurement light from an object. By inputting light (input light) to the light input surface, the light is diffused in the light diffusing part and the light (output light) is emitted from the light output surface. It is characterized in that it is output to irradiate an object with light, and the measurement light from the object transmitted through the light output surface and the back surface is detected.

These light irradiation devices and light measurement devices include the above-mentioned optical plates. Further, these light irradiation methods and light measurement methods use the above-mentioned optical plates. Therefore, for the same reason, it is possible to irradiate light according to desired irradiation conditions.

In the optical measuring device according to the present invention, the imaging device may capture the measurement light emitted from the back surface through the optical plate.

According to the present invention, it is possible to provide an optical plate, a light irradiation device, a light measurement device, a light irradiation method, and a light measurement method capable of irradiating light according to desired irradiation conditions.

It is a schematic cross-sectional view which shows the structure of the optical measuring apparatus which concerns on 1st Embodiment. It is a figure which shows the structure of the optical plate shown in FIG. It is a figure which shows the main process of the method of forming a light diffusing part. It is a figure which shows the main process of the method of forming a light diffusing part. It is a schematic cross-sectional view which shows how light is diffused in a light diffusion part. It is a figure which shows the relationship between the structure of the light diffusing part, and the intensity distribution of the light output from an optical plate. It is a schematic cross-sectional view which shows how the light from a pair of light diffusing parts adjacent to each other is irradiated to the measurement object. It is a figure which shows the simulation result of the light intensity when viewed from the 3rd direction. It is a schematic cross-sectional view which shows how the light diffused in a light diffusing part irradiates an object to be measured. It is a schematic cross-sectional view which shows how light is sequentially diffused by a plurality of light diffusing parts. It is a schematic cross-sectional view which shows the example of the optical plate which further has a reflection part. It is sectional drawing which shows the modification of the optical plate. It is a figure which shows the modification of the optical plate. It is a figure which shows the modification of the optical plate. It is a schematic cross-sectional view which shows the structure of the optical measuring apparatus which concerns on 2nd Embodiment. It is a figure which shows the structure of the optical plate shown in FIG. It is a schematic cross-sectional view which shows how light is diffused in a light diffusion part. It is a schematic cross-sectional view which shows how the light from a plurality of light diffusing parts arranged just below a well is irradiated to a measurement object. It is a figure which shows the structure of the optical measuring apparatus which concerns on 3rd Embodiment. It is a figure which shows the structure of the optical plate shown in FIG. It is a figure which shows the example of the case where a measurement object is held in a petri dish. It is a figure which shows another example when the object to be measured is held in a petri dish. It is a figure which shows the example of the case where the measurement object is held by the slide glass. It is a figure which shows another example when the object to be measured is held on a slide glass.

Hereinafter, an embodiment of an optical plate, a light irradiation device, and a light measurement device according to the present invention will be described in detail with reference to the drawings. In the description of the drawings, the same elements or the corresponding elements may be designated by the same reference numerals, and duplicate description may be omitted. The Cartesian coordinate system S may be shown in the drawings below.
[First Embodiment]

FIG. 1 is a schematic cross-sectional view showing the configuration of the optical measuring device according to the first embodiment. As shown in FIG. 1, the light measuring device 1 according to the first embodiment includes an object holding mechanism 10, a light irradiating device 20, an optical system 30, and an imaging device 40. The object holding mechanism 10 has, for example, a microplate (holding member) 11 and a plate holder 12 that supports the microplate 11.

The microplate 11 includes a main surface 11a, a back surface 11b on the opposite side of the main surface 11a, and a side surface 11c connecting the main surface 11a and the back surface 11b. The back surface 11b is a surface facing the main surface 11a. A plurality of wells 13 arranged in a two-dimensional array along the extending direction of the main surface 11a are formed on the main surface 11a. The number of wells 13 is, for example, 96 or 384. Here, the well 13 is a prismatic recess extending from the main surface 11a toward the back surface 11b.

Therefore, the bottom surface 13s of the well 13 is a plane substantially parallel to the main surface 11a and the back surface 11b. The bottom 14 of the microplate 11 is made of a material such as glass, quartz, and plastic that transmits light. Alternatively, the entire microplate 11 is made of a material such as glass, quartz, and plastic that transmits light. The bottom portion 14 of the microplate 11 is a portion of the microplate 11 on the back surface 11b side of the bottom surface 13s of the well 13. Further, the shape of the bottom surface 13s of the well 13 may be a U-shape, a V-shape, a pyramid shape, or the like.

The microplate 11 holds an object (object) A to be measured. The object A to be measured is housed in the well 13. The object to be measured A is, for example, a cell such as a stem cell or a tissue containing the cell (tissue). The well 13 contains, for example, a solution B such as a culture solution, a fluorescence indicator, and an evaluation compound together with the measurement object A. The object to be measured A is here settled on the bottom surface 13s of the well 13. However, the measurement target A may be a floating cell or the like. In that case, the object A to be measured is suspended in the culture solution in the well 13. The microplate 11 is supported by the plate holder 12 (placed on the plate holder 12) while holding the object A to be measured in this way.

The plate holder 12 supports the periphery of the microplate 11 so that the back surface 11b of the microplate 11 is exposed from the plate holder 12. More specifically, the plate holder 12 supports the outer peripheral portion and the side surface 11c of the back surface 11b so that the region of the back surface 11b facing the well 13 is exposed. The microplate 11 is manually placed on the plate holder 12. Alternatively, when the object holding mechanism 10 has a plate loader (not shown), the microplate 11 is automatically placed on the plate holder 12 by the plate loader.

The light irradiation device 20 includes an optical plate 50 and a light source 21. The optical plate 50 is for irradiating the object A to be measured with light. The light source 21 is for inputting light from the optical input surface of the optical plate 50 to the optical plate 50. That is, the light source 21 outputs the light input to the light input surface of the optical plate 50. Details of the optical plate 50 will be described later. The light source 21 is, for example, an LED (Light Emitting Diode), an LD (Laser Diode), an SLD (Super Luminescent Diode), a lamp (for example, a xenon lamp, etc.) and the like. The light source 21 is not limited to the mode in which the light source 21 is arranged on the light input surface side of the optical plate 50 as described later. That is, the light source 21 may be arranged at an arbitrary position, for example, and may be in a mode in which light is input to the light input surface of the optical plate 50 via a light guide or the like. The light from the light source 21 irradiates the measurement object A held on the microplate 11 via the optical plate 50.

The optical system 30 transmits the measurement light (for example, fluorescence, reflected light, scattered light) from the measurement object A irradiated with light from the light source 21 through the optical plate 50, and forms an image on the image pickup apparatus 40. Therefore, the optical system 30 includes a wavelength selection filter 31 and a lens 32. The wavelength selection filter 31 blocks the transmission of light such as light (excitation light) emitted from the light source 21 to the measurement object A through the optical plate 50, and selectively selects the measurement light from the measurement object A. To Penetrate. The wavelength selection filter 31 may be arranged between the lens 32 and the image pickup apparatus 40.

The wavelength selection filter 31 is, for example, a wavelength band selection filter (bandpass filter), a lowpass filter, a highpass filter, or the like. In the wavelength selection filter 31, for example, the wavelength of fluorescence of the dye that dyes the object A to be measured is selected. For example, in the wavelength selection filter 31, wavelengths of 450 nm and 560 nm are selected when the dye is a voltage-sensitive dye VSP, and wavelengths of 518 nm are selected when the dye is a calcium ion-sensitive dye Fluo-4. Is selected. The wavelength selection filter 31 may be selected so as to transmit the fluorescence wavelengths of various dyes. For example, the wavelength selection filter 31 may be selected to transmit light having a wavelength of, for example, 450 nm or more. The lens 32 is, for example, a camera lens, which collects the measurement light transmitted through the wavelength selection filter 31 and forms an image on the light receiving surface of the image pickup device (imaging device) 40.

The image pickup apparatus 40 captures the measurement light imaged by the lens 32. The image pickup device 40 is, for example, an area image sensor such as a CCD image sensor or a CMOS image sensor. Further, if the imaging device 40 is configured to be able to detect each of the measurement lights from each well 13, for example, a point sensor such as a photomultiplier tube, a photodiode, or an avalanche photodiode may be used.

In the present embodiment, the light irradiation device 20, the optical system 30, and the image pickup device 40 are arranged in this order on the back surface 11b side of the microplate 11. Therefore, the image pickup apparatus 40 is emitted from the measurement object A irradiated with light from the light irradiation apparatus 20, exits from the back surface 11b of the microplate 11, passes through the optical plate 50, and exits from the back surface of the optical plate 50. The measured light is received through the optical system 30 and an image is taken.

Here, FIG. 2 is a diagram showing the configuration of the optical plate shown in FIG. In particular, FIG. 2B is a plan view of the optical plate 50, and FIG. 2A is a cross-sectional view taken along the line IIa-IIa of FIG. 2B. Further, FIG. 2C is a partially enlarged view of FIG. 2A. As shown in FIG. 2, the optical plate 50 has a substantially rectangular plate shape. The optical plate 50 has a main surface 50a and a back surface 50b on the opposite side of the main surface 50a. The back surface 50b is a surface facing the main surface 50a.

The optical plate 50 has a first portion 51 including a main surface 50a and a second portion 52 including a back surface 50b. The first portion 51 and the second portion 52 have a substantially rectangular flat plate shape. The first portion 51 and the second portion 52 may be integrally formed with each other, or may be formed separately from each other and joined to each other. When the first portion 51 and the second portion 52 are formed as separate bodies, they can be fixed to each other by, for example, thin-film deposition or adhesion. As a result, the first portion 51 and the second portion 52 are optically bonded to each other. The dimensions of the first portion 51 in the first direction (here, the X-axis direction) along the main surface 50a are substantially the same as the dimensions of the second portion 52 in the first direction. Therefore, both end faces of the first portion 51 and the second portion 52 in the first direction form the side surface 50c of the optical plate 50 so as to coincide with each other (continuously). However, both end faces of the first portion 51 and the second portion 52 in the first direction do not have to coincide with each other.

On the other hand, the dimensions of the first portion 51 in the second direction along the main surface 50a (the direction intersecting the first direction, here the Y-axis direction) are the dimensions of the second portion 52 in the second direction. Smaller than the dimensions. Therefore, both end faces of the first portion 51 and the second portion 52 in the second direction do not coincide with each other and form the side surfaces 50d and 50f of the optical plate 50. The side surface 50d is the end face of the first portion 51, and the side surface 50f is the end face of the second portion 52. The side surface 50f is a surface that is substantially parallel to the side surface 50d and projects outward from the side surface 50d of the optical plate 50. These side surfaces 50c, 50d, 50f are surfaces between the main surface 50a and the back surface 50b. However, both end faces of the first portion 51 and the second portion 52 in the second direction may form the side surfaces of the optical plate 50 so as to coincide with each other (continuously).

As shown in FIG. 1, here, the light source 21 is arranged so as to be in contact with the side surface 50f. Therefore, in the optical plate 50, the side surface 50f is an optical input surface for inputting light (input light) into the optical plate 50. However, the side surface 50d or the side surface 50c may be used as the optical input surface. On the other hand, the optical plate 50 is arranged so that its main surface 50a faces the measurement object A (the back surface 11b of the microplate 11) held by the microplate 11. Therefore, in the optical plate 50, the main surface 50a facing the measurement object A serves as an optical output surface for outputting light (output light) from the optical plate 50 toward the measurement object A.

Here, a plurality of light diffusing portions 60 are formed on the optical plate 50. Here, the light diffusing portion 60 is formed inside the optical plate 50. In particular, the light diffusing portion 60 is formed in the first portion 51 and not in the second portion 52. That is, the first portion 51 is a portion where the light diffusing portion 60 is formed, and the second portion 52 is a portion where the light diffusing portion 60 is not formed. The portion where the light diffusing portion 60 is not formed is, for example, a portion where the light diffusing portion 60 does not exist between the end faces facing each other. The light diffusing unit 60 causes the light input into the optical plate 50 from the side surface 50f to be output to the outside of the optical plate 50 from the main surface 50a, which is the light output surface, while diffusing (for example, scattering or refracting). That is, the light diffusing unit 60 is for diffusing light inside the optical plate 50. On the other hand, the second portion 52 functions as a light guide unit that guides light without diffusing light between a pair of side surfaces 50f and 50f facing each other. That is, the light guide portion is configured in the second portion 52. By providing a portion in which the light diffusing portion 60 does not exist between the end faces facing each other in this way, the light is emitted from the light output surface so that the light intensity along the light output surface (here, the main surface 50a) becomes substantially uniform. Can be output.

The light diffusing portion 60 extends along the first direction (X-axis direction) between the pair of side surfaces 50c and 50c. Further, the light diffusing portions 60 are arranged at substantially equal intervals along the second direction (Y-axis direction). The spacing between the arrangements of the light diffusing portions 60 is defined according to the object A to be measured, and is defined by, for example, the dimensions and arrangement of the wells 13 in the second direction. More specifically, the light diffusing unit 60 alternates with the well 13 (so as to sandwich the well 13) when viewed from a third direction (here, the Z-axis direction) intersecting the first and second directions. Is arranged. Therefore, most of the light diffusing portions 60 are arranged so as to be located between the wells 13 adjacent to each other when viewed from the third direction. In other words, the microplate 11 is arranged so that the light diffusing portion 60 has such an arrangement.

Here, the light diffusing portion 60 extends along the third direction as well. More specifically, the light diffusing portion 60 extends from the main surface 50a toward the back surface 50b with a predetermined length in the thickness direction (third direction) of the optical plate 50. That is, the light diffusing portion 60 is formed so as to extend over a part between the main surface 50a and the back surface 50b. Such a light diffusing portion 60 is formed by condensing laser light on a member (hereinafter, may be referred to as a “processed object”) that is a source of the optical plate 50.

An example of a method of forming the light diffusing portion 60 will be described. The specific procedure for forming the light diffusing portion 60 is not limited to the following examples. First, as shown in FIG. 3A, a work target 70 as a base of the optical plate 50 (or the first portion 51) is prepared. A plurality of (for example, virtual) scheduled processing lines 5 are set in the processing object 70 so as to correspond to the positions where the light diffusing portion 60 is formed. Subsequently, as shown in FIG. 3B, for example, the main surface 50a of the object to be processed 70 is used as the incident surface of the laser beam L, and the laser beam is emitted at a predetermined position (predetermined depth) in the third direction. The light collecting point P of L is positioned. The condensing point P is a point where the laser beam L condenses.

In that state, the condensing point P of the laser beam L is moved in the first direction relative to the object to be machined 70 along the scheduled machining line 5 (that is, the laser beam L is scanned in the first direction). Let). As a result, as shown in FIG. 3C, a row of modified regions 7 along the first direction is formed inside the workpiece 70. The modified region 7 constitutes, for example, a light diffusing portion 60 (first light diffusing portion 61 in FIG. 2) located on the outermost side (side surface 50d side) of the optical plate 50 in the second direction.

Subsequently, as shown in FIG. 4A, the position of the focusing point P of the laser beam L in the third direction is maintained at a predetermined position (predetermined depth) in the second direction. The position of the focusing point P of the laser beam L is shifted. In this state, as shown in FIG. 4B, the laser beam L is scanned again in the first direction along another scheduled processing line 5 (the focusing point P is relatively moved). As a result, as shown in FIG. 4C, another row of modified regions 7 along the first direction is formed. The modified region 7 constitutes, for example, a light diffusion unit 60 (second light diffusion unit 62 in FIG. 2) adjacent to the first light diffusion unit 61.

In this way, while changing the position of the condensing point P of the laser beam L in the second direction (without changing the third direction), the laser beam L is directed in the first direction along each scheduled processing line 5. Scan for. Then, after the scanning of the laser beam L along all the scheduled processing lines 5 is completed, the position of the focusing point P of the laser beam L in the third direction is set to the main surface 50a side (that is, the incident surface of the laser beam L). Shift to the side) and repeat the same process. As a result, as shown in FIG. 2, the light diffusing portion 60 composed of a plurality of rows of modified regions 7 arranged along the third direction extends along the first direction, and , A plurality are formed so as to be arranged along the second direction. As a result, the optical plate 50 is manufactured.

Here, the modified region 7 may be formed continuously or intermittently. Further, the modified region 7 may be in a row shape or a dot shape. The modified region 7 may be formed at least inside the object to be processed 70. Further, cracks may be formed starting from the modified region 7, and the cracks and the modified region 7 may be exposed on the outer surface (for example, the main surface 50a) of the object to be processed 70.

Further, the laser beam L here is transmitted through the object to be processed 70 and is particularly absorbed in the vicinity of the condensing point P inside the object to be processed 70, whereby the modified region 7 is formed in the object to be processed 70. Is done (ie, internally absorbed laser machining). Therefore, since the laser beam L is hardly absorbed by the main surface 50a of the object to be processed 70, the main surface 50a of the object to be processed 70 does not melt. Generally, when the main surface 50a is melted and removed to form a removal portion such as a hole or a groove (front surface absorption type laser machining), the machining region gradually progresses from the main surface 50a side to the back surface 50b side.

The modified region 7 formed in the present embodiment refers to a region in which the density, refractive index, mechanical strength and other physical characteristics are different from those of the surroundings. The modified region 7 includes, for example, a melt processing region, a crack region, a dielectric breakdown region, a refractive index change region, and the like, and there is also a region in which these are mixed. Further, the modified region 7 includes a region in which the density of the modified region 7 is changed in comparison with the density of the non-modified region in the material of the object to be processed 70, and a region in which lattice defects are formed (these are these). Collectively referred to as the dense metastatic region).

In addition, the melt-treated region, the refractive index change region, the region where the density of the modified region changes as compared with the density of the non-modified region, and the region where lattice defects are formed are further inside those regions and the modified region. Cracks (cracks, microcracks, etc.) may be included at the boundary between 7 and the non-modified region. The included cracks may extend over the entire surface of the modified region 7, or may be formed only in a part or in a plurality of parts.

Further, in the present embodiment, the modification region 7 is formed by forming a plurality of modification spots (processing marks) along the scheduled processing line 5. A reforming spot is a reforming portion formed by a one-pulse shot of pulsed laser light (that is, one-pulse laser irradiation: laser shot), and is a reforming region formed by gathering the reforming spots (or independently). It becomes 7. Examples of the modified spot include crack spots, melt-treated spots, refractive index change spots, and spots in which at least one of these is mixed. The modified region 7 shown in the cross section of FIG. 2 (c), FIG. 3 (c), and FIG. 4 (c) is, for example, this modified spot.

The modified region 7 may be formed linearly by combining linear modified spots extending along the first direction. In that case, by using the laser beam L of the beam spot having a long axis at the condensing point P, a linear modified spot along the long axis can be formed. Further, by adjusting the direction of the long axis of the beam spot of the laser beam L at the condensing point P, the extending direction of the linear modified spot can be adjusted.

An example of the laser processing conditions for forming the modified region 7 (that is, the light diffusing portion 60) in the present embodiment is as follows.
Laser light source: Semiconductor laser excitation Yb: KGW laser Wavelength of laser light L: 1030 nm
Repeat frequency of laser light L: 50 kHz
Pulse width of laser light L: 350 fs
Laser light L output: 10 μJ / pulse

FIG. 5 is a schematic cross-sectional view showing how light is diffused in the light diffusing portion. In FIG. 5, the optical path is indicated by an arrow. Further, in FIG. 5, for ease of explanation, a case where the light diffusing portion 60 is composed of a single modified region 7 (modified spot) in the third direction is illustrated. As shown in FIG. 5, the light input into the optical plate 50 and reaching the light diffusing unit 60 is diffused in each direction by the light diffusing unit 60.

Of the light diffused by the light diffusing unit 60, the light incident on the main surface 50a at a relatively small angle is emitted from the main surface 50a while being refracted on the main surface 50a. The light emitted from the main surface 50a propagates through the intermediate layer M (air layer as an example) between the microplate 11 and the optical plate 50, and is incident on the microplate 11 from the back surface 11b of the microplate 11. The light incident on the microplate 11 irradiates the measurement object A held in the well 13. Then, the measurement light (fluorescence, reflected light, scattered light, etc.) generated by irradiating the measurement object A with light passes through the main surface 50a and the back surface 50b, and is transmitted by the image pickup apparatus 40. Detected.

Here, it is assumed that the refractive index of the intermediate layer M is smaller than the refractive index of the optical plate 50. Therefore, the light diffused in the light diffusing portion 60 is refracted so as to be diffused more widely between the main surface 50a and the intermediate layer M (that is, refracted so that the angle of the well 13 with respect to the depth direction becomes large). Then, the object to be measured A is irradiated. On the other hand, among the light diffused by the light diffusing unit 60, the light incident on the main surface 50a at a relatively large angle is totally reflected by the main surface 50a and is not emitted from the main surface 50a. The light totally reflected on the main surface 50a propagates in the optical plate 50 and is diffused again in another light diffusing unit 60. When the material of the optical plate 50 is synthetic quartz having a refractive index of about 1.464 and the material of the intermediate layer M is air having a refractive index of about 1.00, total reflection occurs on the main surface 50a. The resulting critical angle is about 43 degrees.

Here, an embodiment of the light irradiation method and the light measurement method will be described. In the light irradiation method according to the present embodiment, the optical plate 50 is used as described above. That is, in the light irradiation method according to the present embodiment, the main surface 50a (light output surface) for outputting light, the back surface 50b facing the main surface 50a, and the surface between the main surface 50a and the back surface 50b. An optical plate 50 including a side surface 50f (light input surface) into which light is input and a light diffusing portion 60 formed by condensing the laser beam L is used on the measurement object A (object). Irradiate with light.

More specifically, in the light irradiation method according to the present embodiment, first, light is input to the side surface 50f using the light source 21, and light is input from the side surface 50f into the optical plate 50. The light input into the optical plate 50 guides the inside of the optical plate 50. The light that guides the inside of the optical plate 50 and reaches the light diffusing unit 60 is diffused in the light diffusing unit 60. Then, at least a part of the light diffused by the light diffusing unit 60 (light incident on the main surface 50a at a relatively small angle) is output from the main surface 50a and irradiated to the measurement object A.

That is, in the light irradiation method according to the present embodiment, the light input step of inputting light to the side surface 50f, the light diffusion step of diffusing the light input from the side surface 50f in the light diffusing unit 60, and the diffusing in the light diffusing unit 60. The light output step of outputting the light output from the main surface 50a and the light irradiation step of irradiating the measurement object A with the light output from the main surface 50a are included. Further, the light irradiation method according to the present embodiment can include an optical light guide step that guides the light input from the side surface 50f in the optical plate 50.

Further, in the light measurement method according to the present embodiment, after the light irradiation method described above, the measurement light (fluorescence, reflected light, scattered light, etc.) from the measurement object A irradiated with the light from the optical plate 50 is further applied. Is detected (imaging) by the imaging device 40. In particular, here, the measurement light from the measurement object A that has passed through the main surface 50a and the back surface 50b is detected. That is, the light measurement method according to the present embodiment further includes a light detection step for detecting the measurement light from the measurement object A transmitted through the main surface 50a and the back surface 50b, in addition to each step of the light irradiation method described above. ..

FIG. 6 is a diagram showing the relationship between the configuration of the light diffusing portion and the intensity distribution of the light output from the optical plate. FIG. 6A shows a case where the light diffusing portion 60 is composed of a single modified region 7 (modified spot) in the third direction as shown in FIG. Further, FIG. 6B shows a case where the light diffusing portion 60 is composed of a plurality of modified regions 7 (modified spots) in the third direction as shown in FIG. 2C. That is, FIG. 6B shows a case where the light diffusing portion 60 extends from the main surface 50a toward the back surface 50b with a predetermined length in the thickness direction (third direction) of the optical plate 50. Shown.

As shown in FIG. 6A, when the light diffusing section 60 is composed of a single modified region 7 in the third direction, it corresponds to the light diffusing section 60 (in the second direction). The intensity distribution becomes such that it attenuates as the distance from the center position increases. On the other hand, as shown in FIG. 6B, when the light diffusing portion 60 is composed of a plurality of modified regions 7 in the third direction, the light diffusing portion 60 starts from one modified region 7. Light directed in the third direction is diffused again as it passes through another modified region 7. Therefore, in this case, the intensity decreases at the center position (in the second direction) corresponding to the light diffusing portion 60, but the intensity on both sides thereof can be improved. That is, by adjusting the length of the light diffusing portion 60 extending in the third direction from the main surface 50a to the back surface 50b, the central position (in the second direction) corresponding to the light diffusing portion 60 and its center position thereof. The light intensity on both sides can be adjusted.

FIG. 7 is a schematic cross-sectional view showing how light from a pair of light diffusing portions adjacent to each other irradiates the object to be measured. As shown in FIGS. 2 and 7, here, the light diffusing portions 60 are arranged on both sides of the well 13 when viewed from the third direction. Therefore, the light L1 held in the well 13 is diffused by one light diffusing portion 60 and output from the main surface 50a of the optical plate 50, and is diffused by the other light diffusing portion 60. The light L2 output from the main surface 50a of the optical plate 50 is superposed on each other and irradiated. In other words, the object A to be measured can be illuminated by the lights L1 and L2 from the outside of the well 13 when viewed from the third direction. Therefore, oblique illumination (that is, illumination by a dark field) can be preferably performed on the measurement object A.

FIG. 8 is a diagram showing a simulation result of the light intensity distribution when viewed from the third direction. In FIG. 8, the magnitude of the light intensity is shown by the shade of color. FIG. 8A is a diagram showing the distribution of the light intensity output from the optical plate 50, and FIG. 8B is a diagram showing the light intensity in a state where the microplate 11 is arranged on the optical plate 50. It is a figure which shows the distribution. As shown in FIG. 8, according to the optical plate 50 according to the present embodiment, it is possible to irradiate a plurality of wells 13 with light having a substantially uniform intensity.

As described above, the light measuring device 1 according to the present embodiment includes a light irradiating device 20 having an optical plate 50. The optical plate 50 includes a light diffusing unit 60 that diffuses the light input from the side surface 50f (light input surface) and outputs the light from the main surface 50a (light output surface). Therefore, if the optical plate 50 is arranged so that its main surface 50a (light output surface) faces the measurement object A, the light diffused by the light diffusing unit 60 can uniformly irradiate the measurement object A. It will be possible. In particular, in the optical plate 50, the light diffusing portion 60 is formed by condensing the laser beam L on the member (processed object 70) to be the optical plate 50 (that is, by laser processing). Therefore, the light diffusing portion 60 can be easily formed, and light can be irradiated according to the irradiation conditions such as the shape of the object A to be measured (for example, the shape, dimensions, and arrangement of the well 13).

Further, in the light measuring device 1 according to the present embodiment, as shown in FIG. 9, a light diffusing portion 60 extending along a third direction intersecting (substantially orthogonal to) the main surface 50a of the optical plate 50. Is formed on the optical plate 50. Therefore, the amount of light output from the region adjacent to the light diffusing unit 60 can be increased. The region adjacent to the light diffusing portion 60 is, for example, when the light diffusing portion 60 is formed in a straight line so as to be located between the wells 13 as described above when viewed from the third direction. It is a region on both sides of the light diffusing portion 60 that overlaps the well 13. Alternatively, the region adjacent to the light diffusing portion 60 means, for example, when the light diffusing portion 60 is formed so as to surround the well 13 as described later when viewed from a third direction, the light overlapping the well 13 It is an annular region on the outside of the diffuser 60. The light diffusing portion 60 is arranged so as to sandwich (enclose) the well 13 when viewed from the third direction. Then, the lights L1 and L2 input to the optical plate 50 are diffused by these light diffusing portions 60 and then output from the main surface 50a of the optical plate 50, and are output from the bottom surface 13s of the well 13 of the microplate 11 to the well 13. Incident.

Therefore, the lights L1 and L2 that enter the well 13 are incident on the well 13 at an incident angle smaller than 90 degrees with respect to the bottom surface 13s of the well 13. Further, the lights L1 and L2 illuminate the bottom surface 13s of the well 13 relatively strongly. In other words, the light L1 and L2 mainly irradiate the measurement object A settled on the bottom surface 13s of the well 13. Therefore, the background light noise caused by the light from the optical plate 50 irradiating the solution B in the well 13 is reduced.

Here, the optical plate 50 includes a second portion 52 that functions as a light guide portion that guides light between the side surfaces (light input surfaces) 50f and 50f that face each other. Therefore, the light input from the side surface 50f is diffused and output by the light diffusing unit 60 while being guided between the side surfaces 50f and 50f by the second portion 52 as the light guide unit. Therefore, it is possible to more uniformly irradiate the measurement object A existing in various places on the region where the optical plate 50 is arranged.

Further, the optical plate 50 is composed of a first portion 51 in which the light diffusing portion 60 is formed and a second portion 52 integrally formed with the first portion 51 and in which the light diffusing portion 60 is not formed. At the same time, a light guide portion may be formed in the second portion 52 thereof. In this case, if the laser machining is controlled so that the light diffusing portion 60 is not formed in the second portion 52 of the first portion 51 and the second portion 52 integrally configured with each other, the second portion thereof. The light guide portion can be configured by 52. That is, in this case, the light guide portion can be configured in the optical plate 50 by controlling the laser processing.

Alternatively, the optical plate 50 has a first portion 51 in which the light diffusing portion 60 is formed and a second portion 52 that is formed separately from the first portion 51 and in which the light diffusing portion 60 is not formed. In addition to being formed by optically joining, the light guide portion may be formed by the second portion 52 thereof. In this case, if the first portion 51 in which the light diffusing portion 60 is formed by laser processing and the second portion 52 in which the light diffusing portion is not formed are prepared and optically joined to each other, the second portion 51 thereof. The light guide portion can be formed by the portion 52 of the above.

Here, the action between the light diffusing units 60 will be described. FIG. 10 is a schematic cross-sectional view showing how light is sequentially diffused by a plurality of light diffusing portions. As shown in FIG. 10, the light input to the optical plate 50 is first diffused by the first light diffusing unit 61. The light diffused for the first time in the first light diffusing unit 61 is referred to as primary diffused light. A part of the light L1 of the primary diffused light is continuously diffused in the second light diffusing portion 62 adjacent to the first light diffusing portion 61. The light diffused in the second light diffusing unit 62 in this way is referred to as secondary diffused light. Further, a part of the light L2 of the secondary diffused light is sequentially diffused by the plurality of light diffusing units 60. As an example, the light further diffused in the nth light diffuser 6n is referred to as nth diffused light.

The light output from the main surface 50a of the optical plate 50 and used to illuminate the object A to be measured is affected by the first-order diffused light and the higher-order diffused light from the second-order diffused light to the n-th-order diffused light. Is also added. Therefore, in order to reduce the irradiation unevenness of the object A to be measured, the higher-order diffused light from the second-order diffused light to the n-th-order diffused light is reduced, or the higher-order diffusion such as the second-order diffusion to the nth-order diffusion May be uniformly generated throughout the optical plate 50.

To that end, the optical plate 50 can further have a pair of reflectors 80, as shown in FIG. The reflecting portion 80 extends along a third direction intersecting the main surface 50a, which is the light output surface of the optical plate 50. The reflecting unit 80 reflects at least the light from the light source 21. The reflection unit 80 includes a first reflection region 81 arranged on the side surface 50d of the optical plate 50, and a second reflection region 82 projecting from the main surface 50a in a third direction.

When the reflecting portion 80 has the first reflecting region 81, for example, the light L1 diffused in the predetermined light diffusing portion 60 and incident on the side surface 50d is reflected by the first reflecting region 81 and another light. It will be continuously diffused to the diffusion unit 60. By forming a pseudo light diffusing unit 60 (light diffusing unit 60P) at a position where the reflected light L1 is swept in the first reflection region 81, the light L1 is converted into the pseudo light diffusing unit 60P. Corresponds to the light L2 diffused in. That is, the first reflection region 81 causes pseudo-higher-order diffusion. Therefore, high-order diffusion is uniformly generated in the entire optical plate 50, and it is possible to reduce irradiation unevenness on the measurement object A. In order to form such a first reflection region 81, there are a method of providing a reflection member such as a mirror on the side surface 50d, a method of performing reflection processing on the side surface 50d, and the like.

On the other hand, when the reflecting portion 80 has the second reflecting region 82, for example, the light L3 that is emitted from the main surface 50a after being diffused by the predetermined light diffusing portion 60 and normally deviates from the measurement object A. Is reflected in the second reflection region 82 and irradiates the measurement object A. This creates a pseudo optical plate 50 (optical plate 50P) at a position where the reflected light L3 is swept in the second reflection region 82, and measures the light L4 from the pseudo optical plate 50P. Corresponds to irradiating A. Thereby, the irradiation unevenness can be further reduced. In order to form such a second reflection region 82, there is a method of providing a reflection member such as a mirror so as to project from the main surface 50a in the third direction.

The optical plate 50 described above can be deformed in various ways. FIG. 12 is a cross-sectional view showing a modified example of the optical plate. As shown in FIG. 12A, in the optical plate 50, the light diffusing portion 60 can be formed so as to reach the main surface 50a and not the back surface 50b. That is, the light diffusing portion 60 may be formed at least inside the optical plate 50.

Further, as shown in FIG. 12B, in the optical plate 50, the light diffusing portion 60 is provided only inside the optical plate 50 so as to extend from the first portion 51 to the second portion 52. It may be formed. Also in this case, the light guide portion is formed in the second portion 52. More specifically, in this case, a part of the second portion 52 on the back surface 50b side on which the light diffusing portion 60 is not formed guides light between the pair of side surfaces 50f and 50f facing each other. Functions as an optical part.

Further, as shown in FIG. 12 (c), in the optical plate 50, the light diffusing portion 60 can be formed so as to reach the back surface 50b and not the main surface 50a. In this case, for example, the light source 21 can be arranged on the side surface 50d, which is the end surface of the first portion 51, and the side surface 50d can be used as an optical input surface for inputting light to the optical plate 50. In this case, a part of the first portion 51 on the main surface 50a side on which the light diffusion portion 60 is not formed can function as a light guide portion that guides light between a pair of side surfaces 50d and 50d facing each other. it can. In other words, in this case, the first portion 51 corresponds to the portion formed by the light guide portion (the second portion 52 described above). Further, in this case, the second portion 52 corresponds to the portion where the light diffusing portion 60 is formed (the first portion 51 described above). Further, the back surface 50b can be used as an optical output surface for outputting light.

Further, as shown in FIG. 12D, the formation position in the third direction and the length in the third direction may be randomly set for each light diffusing unit 60.

Subsequently, a modified example of the optical plate 50 will be described. FIG. 13 is a diagram showing a modified example of the optical plate. FIG. 13A is a cross-sectional view taken along the line XIIIa-XIIa in the plan view of FIG. 13B. Further, FIG. 13 (c) is a cross-sectional view taken along the line XIIIc-XIIIc in the plan view of FIG. 13 (d). As shown in FIG. 13, in the optical plate 50, the light diffusing portion 60 can be formed so as to correspond to the shape of the well 13 of the microplate 11 holding the measurement object A.

For example, when the well 13 of the microplate 11 is a prismatic recess arranged in a two-dimensional array, the outer shape of each well 13 is as shown in FIGS. 13 (a) and 13 (b). The light diffusing portion 60 can be formed in a square tubular shape along the above. Alternatively, when the well 13 of the microplate 11 is a cylindrical recess arranged in a two-dimensional array, the outer shape of each well 13 is as shown in FIGS. 13 (c) and 13 (d). A cylindrical light diffusing portion 60 can be formed along the above. When forming these light diffusing portions 60, the arrangement of the optical plate 50 and the microplate 11 can be set so that each of the light diffusing portions 60 is arranged with respect to each of the wells 13.

FIG. 14 (a) is a cross-sectional view taken along the line XIVa-XIVa in the plan view of FIG. 14 (b). Further, FIG. 14 (c) is a cross-sectional view taken along the line XIVc-XIVc in the plan view of FIG. 14 (d). As shown in FIGS. 14A and 14B, in the optical plate 50, the light diffusing portions 60 may be formed in a grid pattern along the main surface 50a. At this time, when viewed from the third direction, the light diffusing portion 60 along one direction and the light diffusing portion 60 along the other direction are formed so as to pass between the wells 13 adjacent to each other.

Further, at this time, when viewed from the third direction, the light diffusing portion 60 along one direction and the light diffusing portion 60 along the other direction may intersect each other at an angle of 90 degrees. They may intersect at angles other than (for example, 45 degrees). In the above case, by forming the light diffusing portion 60 so as to surround the well 13 when viewed from the third direction, the light from the outside of the well 13 obliquely illuminates the measurement object A (that is, that is). Illumination with a dark field) can be preferably performed. Further, in the optical plate 50, not only when the light diffusing portion 60 is formed linearly along the main surface 50a, but also along the main surface 50a as shown in FIGS. 14 (c) and 14 (d). The light diffusing portion 60 may be formed in a curved shape.

Further, each of the above light diffusing portions 60 is composed of a plurality of light diffusing layers (layers including the modified region 7) in directions along the main surface 50a (that is, in the first and / or second directions). May be good. That is, each of the light diffusing units 60 may be composed of a plurality of light diffusing layers formed by condensing the laser beam L. More specifically, for example, when each of the light diffusing portions 60 has the embodiment shown in FIG. 2, a plurality of modified regions extending along the first direction and arranged in the third direction. A first light diffusing layer containing 7 and another plurality of modified regions 7 formed in close proximity to the first light diffusing layer, extending along the first direction and arranged in the third direction. It may be composed of a second light diffusing layer (s) including.

Further, for example, when the light diffusing portion 60 has the embodiment shown in FIGS. 13 (a) and 13 (b), the first light diffusing layer including the square tubular modified region 7 and the first light diffusing layer. It may be composed of a second light diffusing layer (s) including another modified region 7 having a square tube shape so as to surround the light diffusing layer. Further, for example, when the light diffusing portion 60 has the embodiment shown in FIGS. 13 (c) and 13 (d), the first light diffusing layer including the cylindrical modified region 7 and the first light It may be composed of a second light diffusing layer (s) containing another cylindrical modified region 7 concentric with the diffusing layer.
[Second Embodiment]

Subsequently, the optical measuring device according to the second embodiment will be described. FIG. 15 is a schematic cross-sectional view showing the configuration of the optical measuring device according to the second embodiment. As shown in FIG. 15, the light measuring device 1A according to the second embodiment is different from the light measuring device 1 in that the light irradiating device 20A is provided instead of the light irradiating device 20 and in the positional relationship of each element. doing.

The light irradiation device 20A is different from the light irradiation device 20 in that it has an optical plate 50A instead of the optical plate 50. Details of the optical plate 50A will be described later. The light source 21 inputs light to the optical plate 50A from the optical input surface of the optical plate 50A. The optical system 30 transmits the measurement light (for example, fluorescence, reflected light, scattered light) from the measurement object A irradiated with light from the light source 21 through the optical plate 50, and forms an image on the image pickup apparatus 40. The image pickup apparatus 40 captures the measurement light imaged by the lens 32 of the optical system 30.

In the present embodiment, the light irradiation device 20A is arranged on the back surface 11b side of the microplate 11, and the optical system 30 and the image pickup device 40 are arranged on the main surface 11a side of the microplate 11 in this order. Therefore, the image pickup apparatus 40 receives the measurement light emitted from the measurement object A irradiated with the light from the light irradiation apparatus 20A and emitted from the main surface 11a side of the microplate 11 via the optical system 30 for imaging. To do.

FIG. 16 is a diagram showing the configuration of the optical plate shown in FIG. In particular, FIG. 16A is a plan view of the optical plate 50A, and FIG. 16B is a partial cross-sectional view of the optical plate 50A. As shown in FIG. 16, the optical plate 50A differs from the optical plate 50 in the mode of forming the light diffusing portion 60. More specifically, in the optical plate 50A, the light diffusing portion 60 extends along the first direction (X-axis direction) between the pair of side surfaces 50c and 50c.

Further, the light diffusing portions 60 are arranged at substantially equal intervals along the second direction (Y-axis direction). The spacing between the arrangements of the light diffusing units 60 is set so that the wells 13 and the light diffusing units 60 in a plurality of rows overlap when viewed from the third direction (Z-axis direction). Therefore, when viewed from the third direction, a plurality of rows of light diffusing portions 60 are arranged directly below one well 13. On the other hand, here, the light diffusion unit 60 is composed of one modification region 7 (modification spot) in the third direction. However, the light diffusing unit 60 may be composed of a plurality of modified regions 7 in the third direction as in the first embodiment. The method of forming the light diffusing portion 60 is the same as that of the first embodiment.

FIG. 17 is a schematic cross-sectional view showing how light is diffused in the light diffusing portion. In the first embodiment, the intermediate layer M between the microplate 11 and the optical plate 50 is made of a material (for example, air) smaller than the refractive index of the optical plate 50. In the present embodiment, it is assumed that the intermediate layer M between the microplate 11 and the optical plate 50A is made of a material having a refractive index higher than that of the material of the optical plate 50. As an example of the intermediate layer M in this case, for example, a resin such as silicone, oil, water, or the like is arranged between the main surface 50a of the optical plate 50A and the back surface 11b of the microplate 11. Can be mentioned.

Therefore, the light diffused in the light diffusing portion 60 is refracted between the main surface 50a and the intermediate layer M so that the degree of diffusion is small (that is, the angle of the well 13 with respect to the depth direction is small). Refracted) and emitted from the main surface 50a. As described above, the light diffusing unit 60 is also arranged directly below the well 13. Therefore, the measurement object A is sufficiently illuminated even if the light has a relatively small spread.

FIG. 18 is a schematic cross-sectional view showing how light from a plurality of light diffusing portions arranged directly under the well is applied to the measurement object. As shown in FIG. 18, here, the lights L1 and L2 diffused in the light diffusing portions 60 adjacent to each other are alternately superposed and irradiated to the measurement object A held in the well 13. In other words, the object A to be measured can be illuminated by the lights L1 and L2 from directly below the well 13 when viewed from the third direction. Therefore, transmitted illumination (that is, illumination with a bright field) can be preferably performed on the object A to be measured.

As described above, the light measuring device 1A according to the present embodiment can also irradiate the light according to the desired irradiation conditions as in the light measuring device 1 according to the first embodiment. In particular, in the light measuring device 1A, the light is diffused by the plurality of light diffusing portions 60 arranged directly under the well 13 in a state where the angle with respect to the depth direction (third direction) of the well 13 is relatively small. The measurement object A held in the well 13 is irradiated. Therefore, illumination with a bright field can be preferably performed.

Various modifications of the optical plate 50 according to the first embodiment are also effective for the optical plate 50A according to the present embodiment. That is, the light diffusing portion 60 may be formed in a square tube shape or a cylindrical shape in a range in which a plurality of light diffusing portions 60 are arranged for one well 13 when viewed from the third direction. The light diffusing portion 60 may be formed in a grid pattern or a curved shape along the surface 50a. Alternatively, the formation position and length of the light diffusing portion 60 in the third direction may be appropriately set. Further, the reflecting unit 80 may be further provided.
[Third Embodiment]

Subsequently, the optical measuring device according to the third embodiment will be described. FIG. 19 is a diagram showing a configuration of an optical measuring device according to a third embodiment. In particular, FIG. 19A is a schematic cross-sectional view taken along the line XIXa-XIXa in the plan view of FIG. 19B, and FIG. 19C is a side view. As shown in FIG. 19, the light measuring device 1B according to the third embodiment is different from the light measuring device 1 in that the light irradiating device 20B is provided in place of the light irradiating device 20. In FIG. 19, the optical system 30 and the image pickup apparatus 40 are omitted.

The light irradiation device 20B is different from the light irradiation device 20 in that it has an optical plate 50B instead of the optical plate 50. The optical plate 50B has a pair of reflecting portions 90. The reflecting portion 90 extends along a third direction intersecting the main surface 50a, which is the light output surface of the optical plate 50B. The reflecting portion 90 is provided on the side surface 50c intersecting the side surface 50f, which is the light input surface of the optical plate 50B. The reflection unit 90 has the same effect as the first reflection region 81 of the reflection unit 80 described above.

A mask 97 is provided on the main surface 50a of the optical plate 50B. The mask 97 has a rectangular ring shape when viewed from the third direction, and is provided on the outer peripheral portion of the main surface 50a. Therefore, only the portion of the main surface 50a excluding the outer peripheral portion is exposed from the opening 97h of the mask 97. The dimension of the opening 97h of the mask 97 is defined so that the entire region where the well 13 of the microplate 11 is formed is included in the exposed portion from the opening 97h of the main surface 50a when viewed from the third direction. To.

Further, the optical plate 50B has a reflecting portion 95 erected on the mask 97. The reflecting portion 95 extends along a third direction intersecting the main surface 50a, which is the light output surface of the optical plate 50B. The reflecting portion 95 has a rectangular ring shape along the outer edge of the opening 97h of the mask 97 when viewed from the third direction. The reflection unit 95 has the same effect as the second reflection region 82 of the reflection unit 80 described above.

Here, as shown in FIGS. 19 and 20, the optical plate 50B has a plurality of dot-shaped light diffusing portions 60 arranged in a two-dimensional array along the main surface 50a when viewed from the third direction. Have. Each light diffusing unit 60 is composed of a single modified region 7 (modified spot) in the first and second directions. On the other hand, each light diffusing unit 60 is composed of a plurality of (two in this case) modified regions 7 (modified spots) in the third direction. The method of forming the light diffusing portion 60 is the same as that of the first embodiment.

The arrangement of the light diffusing unit 60 when viewed from the third direction is defined as follows. That is, as shown in FIG. 20 (b), the ratio of the area of the light diffusing unit 60 to the area of a single pixel PA of the image pickup device (image sensor) 40 is less than 10% when viewed from the third direction. The dimensions, arrangement, and the like of the light diffusing unit 60 are defined so as to be. For example, when the pixel PA of the image pickup apparatus 40 is about 0.25 mm × 0.25 mm, if one light diffusing portion 60 having about 0.02 mm × 0.02 mm is arranged in the pixel PA, the area ratio will be increased. , About 0.64%. If the processed area of the light diffusing unit 60 is smaller than the field of view size in this way, the light diffusing unit 60 does not appear as an image at the time of imaging by the imaging device 40, and it is possible to avoid imaging unevenness.

As described above, the light measuring device 1B according to the present embodiment can also irradiate the light according to the desired irradiation conditions as in the light measuring device 1 according to the first embodiment. Further, as described above, it is possible to avoid imaging unevenness.

The embodiment described above describes one embodiment of the optical plate, the light irradiation device, and the light measurement device according to the present invention. Therefore, the optical plate, the light irradiation device, and the light measurement device according to the present invention are not limited to those described above.

For example, in the first to third embodiments, the case where the measurement object A is held in the well 13 of the microplate 11 has been described. However, in the optical plate according to the present invention (that is, a light irradiation device, a light measurement device, a light irradiation method, and a light measurement method), the measurement object A is held by another holding member such as a petri dish or a slide glass. It is also applicable in some cases.

FIG. 21 is a diagram showing an example when the object to be measured is held in a petri dish. 21 (b) is a cross-sectional view taken along the line XXIb-XXIb of FIG. 21 (a). As shown in FIG. 21, here, the object A to be measured is held by the petri dish (holding member) 16, and the optical plate 50C is used accordingly. A plurality of light diffusing portions 60 arranged in an annular shape are formed on the optical plate 50C so as to surround the petri dish 16 when viewed from the third direction. That is, the light diffusing portion 60 is not formed directly under the petri dish 16. Further, each light diffusing unit 60 is composed of a plurality of modified regions 7 (modified spots) in the third direction. That is, the light diffusing portion 60 extends along the third direction.

Also in this case, as in the above embodiment, it is possible to irradiate light according to desired irradiation conditions. Further, since the measurement object A can be illuminated by the light from the outside of the petri dish 16 when viewed from the third direction, oblique illumination (that is, illumination by a dark field) to the measurement object A can be preferably performed. it can.

FIG. 22 is a diagram showing another example when the object to be measured is held in a petri dish. 22 (b) is a cross-sectional view taken along the line XXIIb-XXIIb of FIG. 22 (a). As shown in FIG. 22, here, the object A to be measured is held by the petri dish (holding member) 16, and the optical plate 50D is used accordingly. A plurality of light diffusing portions 60 are formed on the optical plate 50D along the main surface 50a. More specifically, the optical plate 50D is formed with a plurality of light diffusing portions 60 so as to overlap the petri dish 16 when viewed from the third direction. Therefore, when viewed from the third direction, a plurality of light diffusing portions 60 are arranged directly below the petri dish 16. Each light diffusing unit 60 is composed of a single modified region 7 (modified spot) in the third direction.

Also in this case, as in the above embodiment, it is possible to irradiate light according to desired irradiation conditions. Further, since the measurement object A can be illuminated by the light from directly below the petri dish 16 when viewed from the third direction, transmitted illumination (that is, illumination by a bright field) for the measurement object A can be preferably performed. ..

FIG. 23 is a diagram showing an example in the case where the object to be measured is held on the slide glass. FIG. 23 (b) is a cross-sectional view taken along the line XXIIIb-XXIIIb of FIG. 23 (a). As shown in FIG. 23, here, the object to be measured A is held by a rectangular plate-shaped slide glass (holding member) 17, and an optical plate 50E is used accordingly. A plurality of light diffusing portions 60 arranged in a rectangular ring shape are formed on the optical plate 50E so as to surround the slide glass 17 when viewed from the third direction. That is, the light diffusing portion 60 is not formed directly under the slide glass 17. Further, each light diffusing unit 60 is composed of a plurality of modified regions 7 (modified spots) in the third direction. That is, the light diffusing portion 60 extends along the third direction.

Also in this case, as in the above embodiment, it is possible to irradiate light according to desired irradiation conditions. Further, since the measurement object A can be illuminated by the light from the outside of the slide glass 17 when viewed from the third direction, oblique illumination (that is, illumination by a dark field) to the measurement object A can be preferably performed. it can.

FIG. 24 is a diagram showing another example when the object to be measured is held on the slide glass. FIG. 24 (b) is a cross-sectional view taken along the line XXIVb-XXIVb of FIG. 24 (a). As shown in FIG. 24, here, the object A to be measured is held by the slide glass (holding member) 17, and the optical plate 50F is used accordingly. A plurality of light diffusing portions 60 are formed on the optical plate 50F along the main surface 50a. More specifically, the optical plate 50F is formed with a plurality of light diffusing portions 60 so as to overlap the slide glass 17 when viewed from the third direction. Therefore, when viewed from the third direction, a plurality of light diffusing portions 60 are arranged directly below the slide glass 17. Each light diffusing unit 60 is composed of a single modified region 7 (modified spot) in the third direction.

Also in this case, as in the above embodiment, it is possible to irradiate light according to desired irradiation conditions. Further, since the measurement object A can be illuminated by the light from directly below the slide glass 17 when viewed from the third direction, it is possible to preferably perform transmitted illumination (that is, illumination with a bright field) for the measurement object A. it can.

As described above, various modes can be assumed for the formation of the light diffusing portion in the optical plate. The mode of forming the light diffusing portion in the optical plate is determined as follows as an example. That is, first, the specifications of the optical system are set. More specifically, the excitation wavelength from the light source and the distance and number of the intermediate layers are set. The intermediate layer here refers to, for example, a layer between the main surface of the optical plate and the back surface of the holding member such as a microplate (for example, the intermediate layer M), and the back surface of the holding member such as the microplate to the holding member. At least one of the layers (eg, the bottom 14 of the microplate 11) up to the mounting surface of the object to be measured (eg, the bottom surface 13s of the well 13). Further, the distance between the intermediate layers is a dimension in the third direction of those layers.

Subsequently, after setting the specifications of the optical system, the specifications of illumination symmetry are set. More specifically, setting of the shape of illumination symmetry (holding member and measurement object) (that is, setting of irradiation conditions), specifications of incident light (for example, light distribution characteristics, bright field or dark field, and irradiation unevenness). (Tolerance level, etc.) and the required amount of light are set.

After that, the refractive index of the optical plate, the refractive index of the intermediate layer, and the shape of the optical plate are set. With the above settings, the mode of the light diffusing portion formed on the optical plate is determined. That is, the positions and patterns of the light diffusing portions formed on the optical plate in the first to third directions are determined. Therefore, the mode of the light diffusing portion in the optical plate is not limited to the above-mentioned specific example, and can be appropriately set according to various conditions.

Further, the object to be illuminated (for example, the object to be measured A) is not limited to cells or a tissue containing cells (tissue), and may be an industrial product such as a semiconductor device or a film. Further, the optical measuring device of the above embodiment may be a microscope device such as an optical microscope or a digital slide scanner. In this case, an objective lens can be further provided as needed.

1 ... Light measuring device, 11 ... Micro plate (holding member), 16 ... Share (holding member), 17 ... Slide glass (holding member), 20, 20A, 20B ... Light irradiation device, 21 ... Light source, 40 ... Imaging device , 50, 50A, 50B, 50C, 50D, 50E, 50F ... Optical plate, 50a ... Main surface (optical output surface), 50b ... Back surface (optical output surface), 50d, 50f ... Side surface (optical input surface), 51 ... 1st part, 52 ... 2nd part, 60, 61, 62 ... Light diffusing part, 80, 90, 95 ... Reflecting part, A ... Measurement object (object), L1, L2, L3, L4 ... Light , L ... Laser light.

Claims (7)

An optical plate for irradiating an object with light, the light input surface for inputting the light, the light output surface for outputting the light, the back surface facing the light output surface, and the said. An optical plate formed at least inside the optical plate and including a light diffusing portion for diffusing the light and outputting the light from the light output surface.
A light source that outputs the light input to the light input surface and
A holding member that holds the object and
An imaging device that captures the measurement light from the object irradiated with the light output from the light output surface, and an imaging device.
With
The light diffusing portion extends in a direction from the light output surface toward the back surface, and is formed at a plurality of positions so as to be separated from each other in a cross section intersecting the light output surface and the light input surface. ,
The light input surface is an end surface of the optical plate and is a surface between the light output surface and the back surface.
The light input from the light input surface is diffused in the light diffusing portion and output from the light output surface.
Each of the light diffusing parts is composed of a plurality of light diffusing layers.
The light output surface is the main surface of the optical plate.
The light diffused by one of the pair of light diffusing portions adjacent to each other and output from the main surface and the light diffused by the other of the pair of light diffusing portions and output from the main surface are combined. , Overlapping each other and irradiating the object,
The imaging device captures the measurement light emitted from the back surface through the optical plate.
An optical measuring device characterized by the fact that. It has a light guide portion that guides the light between the light input surface and the surface facing the light input surface.
The optical measuring device according to claim 1. A first portion in which the light diffusing portion is formed and a second portion integrally formed with the first portion are included.
The light guide portion is formed of the second portion.
The optical measuring device according to claim 2. It is configured by joining the first portion on which the light diffusing portion is formed and the second portion formed separately from the first portion.
The light guide portion is formed of the second portion.
The optical measuring device according to claim 2. The light diffusing portion is formed over at least a part between the light output surface and the back surface.
The optical measuring device according to any one of claims 1 to 4. The first portion where the light diffusing portion is formed and
A side surface between the light output surface and the back surface, which is an end surface of the first portion,
A reflecting portion having a first reflecting region arranged on the side surface and extending along a direction intersecting the light output surface to reflect the light.
Further prepare,
The optical measuring device according to any one of claims 1 to 5. An optical output surface for outputting light, a back surface facing the optical output surface, a surface between the optical output surface and the back surface, an optical input surface to which the light is input, and the light output surface. It extends from the light toward the back surface and is formed at a plurality of positions so as to be separated from each other in a cross section intersecting the light output surface and the light input surface, and outputs from the light output surface while diffusing the light. An optical plate including a light diffusing portion for causing the light diffusing, each of which is composed of a plurality of light diffusing layers, and the light output surface is a main surface of the optical plate. The light diffused by one of the pair of light diffusing portions adjacent to each other and output from the main surface and the light diffused by the other of the pair of light diffusing portions and output from the main surface are combined. , The process of preparing an optical plate that superimposes on each other and irradiates an object,
A step of irradiating the object with the light using the optical plate and detecting the measurement light from the object irradiated with the light is provided.
The optical input surface is an end surface of the optical plate.
In the step of detecting the measurement light, by inputting the light to the light input surface, the light is diffused in the light diffusing portion and the light is output from the light output surface to cause the object to receive the light. It irradiates light and detects the measurement light from the object that has passed through the light output surface and the back surface.
A light measurement method characterized by the fact that.

Images