Sample measuring device, control method for sample measuring device, and program

Abstract

The objective of the present invention is to acquire an optical signal stably in a sample measuring device using a simple configuration, even if variability in scanning speed occurs. According to the present invention, a sample measuring method includes irradiating an array substrate with condensed primary light, detecting secondary light from a spot and outputting a detection signal, relatively changing the irradiation position of the primary light on the array substrate, acquiring information relating to the relative position of the primary light with respect to the array substrate, and converting the detection signal acquired in a prescribed period into digital data, wherein the sample measuring method further includes holding information relating to a target position Pt set in relation to the irradiation position of the primary light, determining that the irradiation position has reached the target position Pt on the basis of the information relating to the relative position, selecting digital data including digital data correspond to the prescribed period from the digital data in accordance with the determination result, and transferring the digital data to a storage means for storing the digital data.

Description

Specimen measurement device, control method for specimen measurement device, and program

[0001] The present disclosure relates to a sample measurement device, a sample measurement method, and a program for measuring a sample.

[0002] Protein array plates or peptide array plates are known, in which a large number of biological substances with peptide bonds, such as proteins or peptides, are immobilized on a substrate. These plates can be used to simultaneously investigate interactions with a large number of biological substances immobilized on the substrate. In other words, these array plates are effective for comprehensively analyzing the interactions between a large number of proteins or peptides and liquid samples derived from living organisms, such as blood, cell extracts, saliva, and interstitial fluid. Such analyses enable the measurement of sample characteristics. Hereinafter, the immobilized protein or peptide sites on the substrate will be referred to as spots.

[0003] A known method for observing spots that have interacted with a specimen involves labeling the spots with a fluorescent probe and identifying the interacting spots. A fluorescent image scanner is also known as a device for observing array plates labeled with fluorescent probes (Patent Document 1). The fluorescent image scanner disclosed in Patent Document 1 includes an illumination optical system, a fluorescence detection optical system, and a two-dimensional scanning system. The illumination optical system focuses and irradiates the array plate with laser light. The fluorescence detection optical system detects the amount of fluorescence emitted from spots labeled with fluorescent probes upon irradiation with the laser light. The two-dimensional scanning system scans the array plate or the optical system two-dimensionally to change the irradiation position of the laser light on the array plate and acquire a captured image reflecting the amount of fluorescence excited by the laser light at the spots. A confocal optical system is used as the fluorescence detection optical system.

[0004] Furthermore, Patent Document 2 discloses a technology for adjusting the phase of a sampling clock for detecting the amount of fluorescence from a spot in accordance with the scanning timing of two-dimensional scanning, thereby acquiring optical signals at equal intervals. In these prior art two-dimensional scanning mechanisms, a piston-crank mechanism or a galvanometer scanner mechanism is used to perform high-speed reciprocal scanning with a laser beam in the main scanning direction. Furthermore, in these prior art technologies, the amount of fluorescence from the spot is converted into an analog signal by a highly sensitive sensor such as a photomultiplier tube, and then converted into optical signal data, which is a digital value, by AD conversion. AD conversion generally requires a predetermined conversion time determined by the clock frequency of the AD converter and peripheral circuits.

[0005] US Patent No. 7,911,670 JP 2018-81136 A

[0006] In conventional fluorescence image scanners, the travel time required for the focusing point of the irradiated laser light to move between points where optical signals are to be acquired is determined taking into account the scanning speed of the two-dimensional scanning mechanism. Specifically, the clock frequencies of the AD converter and peripheral circuits are determined so that AD conversion is completed in a time shorter than the travel time. However, mechanical vibrations occurring when the laser light is scanned back and forth by the two-dimensional scanning mechanism can cause local variations in the scanning speed of the laser light. This results in variations in the scanning time between points where optical signals are to be acquired. As a result, the scanner may pass a point where the next optical signal is to be acquired before AD conversion of the optical signal from the previous point is completed, potentially resulting in an optical signal not being acquired. The likelihood of this problem occurring increases when optical signals are acquired at short acquisition intervals in order to improve the image quality of captured images.

[0007] Possible solutions to this problem include selecting a faster AD converter with a shorter conversion time, or using a more robust scanning mechanism to sufficiently reduce local speed variations. However, implementing these solutions increases the cost and size of the device. On the other hand, slowing the reciprocating scanning speed to account for local speed variations increases the imaging time. Furthermore, increasing the acquisition interval without changing the scanning speed may result in an insufficient number of optical signal data points being acquired, potentially reducing measurement accuracy.

[0008] The present disclosure has been made in consideration of such circumstances, and aims to provide a sample measurement device and a control method for a sample measurement device that have a simple configuration and are capable of stably acquiring optical signal data even when variations in scanning speed occur.

[0009] In order to solve the above problem, a specimen measurement device according to one aspect of the present disclosure is a specimen measurement device that optically measures the characteristics of a specimen using an array substrate on which spots are arranged in an array, and includes: a light irradiation means that irradiates the array substrate with concentrated primary light for a predetermined period of time; a light detection means that detects secondary light from the spots at a predetermined cycle and outputs a detection signal; a scanning means that changes the irradiation position of the primary light on the array substrate; a position acquisition means that acquires information about the irradiation position; a conversion means that converts the detection signal acquired at the predetermined cycle into digital data; a first storage means that stores information about a target position Pt related to the irradiation position; a second storage means that stores the digital data; a determination means that determines whether the irradiation position corresponds to the target position Pt; a memory means that stores the digital data; and a transfer means that, depending on the determination result of the determination means, selects data from the digital data stored in the second storage means that corresponds to the timing at which the irradiation position reaches the target position Pt and transfers the data to the memory means.a light detection step of detecting secondary light from the spots at predetermined intervals and outputting detection signals; a scanning step of changing the irradiation position of the primary light on the array substrate; a position acquisition step of acquiring information regarding the irradiation position; and a conversion step of converting the detection signals acquired at predetermined intervals into digital data; a determination step of determining whether the irradiation position corresponds to the target position Pt; and a transfer step of selecting, from the digital data stored in the second storage means and based on the determination result obtained in the determination step, data corresponding to the timing at which the irradiation position reaches the target position Pt, and transferring the data to the storage means.

[0010] According to one aspect of the present disclosure, a sample measurement device and a method for controlling a sample measurement device are provided that have a simple configuration and are capable of stably acquiring optical signal data even when variations in scanning speed occur.

[0011] FIG. 1 is a diagram showing a schematic configuration of a specimen measurement device according to a first embodiment. FIG. 1 is a diagram explaining a specimen that is a target of the present disclosure, showing a top view of an array plate. FIG. 2 is a diagram explaining a specimen that is a target of the present disclosure, showing a side view of the array plate. FIG. 3 is a diagram showing the internal configuration of a controller according to the first embodiment. FIG. 4 is a diagram showing an operation flow of an imaging process according to the first embodiment. FIG. 5 is a diagram explaining the positional relationship of sub-scanning in the imaging process according to the first embodiment, showing a top view of the array plate. FIG. 6 is a diagram explaining the positional relationship of sub-scanning in the imaging process according to the first embodiment, showing a schematic diagram of a trajectory of excitation light on the array plate. FIG. 7 is a diagram explaining the positional relationship of height scanning in the imaging process according to the first embodiment, showing an example of a side view of the array plate. FIG. 8 is a diagram explaining the positional relationship of height scanning in the imaging process according to the first embodiment, showing an example of a side view of the array plate. FIG. 9 is a diagram showing the functional configuration provided inside a receiving circuit according to the first embodiment. FIG. 10 is a diagram showing the operation timing of an imaging process according to the first embodiment. FIG. 11 is a timing chart relating to the operation of conventional optical signal data acquisition. FIG. 12 is a timing chart relating to the operation of conventional optical signal data acquisition. FIG. 13 is a timing chart relating to the operation of optical signal data acquisition according to the first embodiment. FIG. 14 is a timing chart relating to the operation of optical signal data acquisition according to the first embodiment. 1 is a timing chart relating to the operation of acquiring optical signal data according to an embodiment. FIG. 2 is a timing chart relating to the operation of acquiring optical signal data according to an embodiment. FIG. 3 is a diagram showing an operation flow of height information acquisition processing according to a first embodiment. FIG. 4 is a diagram explaining the positional relationship of height scanning in the height information acquisition processing according to the first embodiment, showing a side view of the array plate. FIG. 5 is a diagram explaining the positional relationship of height scanning in the height information acquisition processing according to the first embodiment, showing a side view of the array plate. FIG. 6 is a diagram explaining the positional relationship of height scanning in the height information acquisition processing according to the first embodiment, showing a plot of the amount of reflected light acquired by an optical sensor during height scanning for each height. FIG. 7 is a diagram showing an operation flow of an imaging processing according to a second embodiment. FIG. 8 is a diagram showing the relationship of acquisition positions of optical signal data according to the second embodiment.FIG. 10 is a diagram for explaining the processing of level conversion according to a third embodiment, showing the relationship between the output voltage of a current-voltage conversion amplifier and the digital value of the output of an AD converter. FIG. 11 is a diagram for explaining the processing of level conversion according to a third embodiment, showing the relationship between input and output from voltage level conversion inside a receiving circuit to an AD converter. FIG. 12 is a diagram for explaining the processing of level conversion according to the third embodiment, showing the relationship between input and output of pixel level conversion in image formation processing. FIG. 13 is a diagram for explaining the relationship between optical signal data acquisition positions according to a fourth embodiment. FIG. 14 is a diagram for explaining the operation of a piston crank mechanism according to an aspect of the present disclosure. FIG. 15 is a diagram showing a first holding means, a second holding means, and a determination means according to the first embodiment. FIG. 16 is a diagram showing a conversion means according to the first embodiment.

[0012] Exemplary embodiments for implementing the present disclosure will be described in detail below with reference to the accompanying drawings. Note that the following embodiments do not limit the invention according to the claims. Although multiple features are described in the embodiments, not all of these multiple features are necessarily essential to the invention, and multiple features may be combined arbitrarily. Furthermore, the dimensions, materials, shapes, and relative positions of components described in the following embodiments are arbitrary and can be changed depending on the configuration of the device to which the present disclosure is applied or various conditions. Furthermore, in the accompanying drawings, the same reference numbers will be used between drawings to indicate identical or functionally similar elements, and redundant explanations will be omitted.

[0013] First Embodiment A first embodiment of the present disclosure, in which the above-described array plate is placed on the analyte measurement device shown in FIG. 1 to perform analyte measurement, will be described below with reference to FIGS. 3 to 11C. Here, a schematic configuration of the analyte measurement device according to the present disclosure will first be described using FIG. 1. The analyte measurement device 1 shown in FIG. 1 measures an array plate 101 on a glass slide on which numerous spots of biological material are immobilized and fluorescently labeled. The analyte measurement device 1 includes an objective optical system, a conversion means, a scanning means, and a detection means. The objective optical system is configured to irradiate excitation light (described below) and receive fluorescence, and the conversion means is configured to convert the received fluorescence into an electrical signal. The scanning means is configured to relatively scan the excitation light across the array plate 101, and the detection means is configured to detect information about the analyte from the received fluorescence. Each of the components will be described below.

[0014] 1, the specimen measurement device 1 includes an objective optical system 129 as an optical system for acquiring a fluorescent image of the array plate 101. The objective optical system 129 is configured so that light emitted from the light source 102 passes through an objective lens and is focused on the sample surface, and secondary light passes through the objective lens again and is focused on an optical sensor 105 (described below), where the light passes through a pinhole and is received. The objective optical system 129 shares coaxially arranged elements and includes a light irradiating means 122 that irradiates the array plate 101 with concentrated primary light for only a predetermined period, and a light detecting means 125 that detects secondary light from the spot at a predetermined cycle and outputs a detection signal.

[0015] <Light Irradiation Unit> The light irradiation unit 122 includes a light projection unit 104 having an objective lens that irradiates the array plate 101 with concentrated primary light, a confocal optical system 103 having a beam splitter and a pinhole (not shown), and a light source 102. Note that the light projection unit 104 may also be referred to as an objective lens, a light emission unit, a primary light emission unit, a primary light emission unit, or the like.

[0016] <Light Detecting Means> The light detecting means 125 includes a light projecting unit 104 that collects secondary light from the spot, a confocal optical system 103, and a light sensor 105. The light sensor 105 is composed of a detection element that can detect the wavelength of the secondary light with high sensitivity and a wide dynamic range, and a photomultiplier tube (PMT), a semiconductor sensor, or the like is used.

[0017] The light source 102 is a semiconductor laser that emits light with wavelengths of approximately 670 nm and 780 nm (hereinafter referred to as excitation light). In the specimen measurement device 1, the emission and stopping of light at each wavelength can be independently controlled. The confocal optical system 103 guides the excitation light from the light source 102 to the array plate 101 and guides the fluorescence from the spots on the array plate 101 and the reflected light from the surface of the glass slide to the optical sensor. In this example, the confocal optical system 103 is composed of a pinhole, a filter, a dichroic mirror, a quarter-wave plate, a polarizing beam splitter, and a lens. The use of the confocal optical system 103 reduces the influence of autofluorescence from the glass slide and improves the signal-to-noise ratio when measuring the fluorescence components from the spots. The light projector 104 is used to focus and irradiate the excitation light onto the spot on the array plate 101. It includes a prism for directing the excitation light toward the array plate 101 and a lens for focusing the excitation light onto the spot on the array plate 101. The configurations of the confocal optical system 103 and the light projecting unit 104 are not limited to those exemplified here, and may be replaced with various known configurations as long as equivalent functions are obtained.

[0018] The optical sensor 105 is provided to convert the fluorescence into an electrical signal and includes a photomultiplier tube for acquiring the fluorescence and a photodiode for acquiring the reflected light. The optical sensor 105 is capable of separately acquiring the fluorescence from the spots on the array plate 101 and the reflected light from the surface of the array plate 101. The optical sensor 105 is also configured with a plurality of photomultiplier tubes and photodiodes corresponding to the plurality of wavelengths of the light source 102.

[0019] <Conversion Means> As shown in Figure 18, the specimen measurement device 1 includes conversion means 146 that converts detection signals acquired at a predetermined cycle into digital data. The conversion means 146 is composed of an AD converter 705 arranged in the receiving circuit 106 and an AD converter control circuit 306 that controls the receiving circuit 106. As shown in Figure 18 or Figure 7 described below, the receiving circuit 106 is a receiving circuit that amplifies detection signals from a photomultiplier tube 701 provided in the optical sensor 105 and converts them into digital values. The receiving circuit 106 is composed of an amplifier circuit (current-voltage conversion amplifier 702), a level conversion circuit (level conversion circuit 703), a filter circuit (filter circuit 704), an AD converter (ADC 705), and the like.

[0020] <Scanning Means> As shown in Figure 1, the specimen measurement device 1 includes a scanning means 130 that changes the irradiation position Pi of the primary light on the array plate 101. The scanning means 130 includes a mechanism for two-dimensionally scanning the light projecting unit 104 across the array plate 101, and includes a main scanning means 130x that moves the light projecting unit 104 in the X direction in the two-dimensional direction, and a sub-scanning means 130y that moves the array plate 101 in the Y direction. The main scanning means 130x includes a pulse motor 107, a linear stage 109, a motor driver 115, and a piston crank mechanism 120. The piston crank mechanism 120 includes a crank 118 and a connecting rod 119. The sub-scanning means 130y includes a linear stage 112y, a pulse motor 113, and a motor driver 114. The scanning means 130 further includes a z-direction scanning means 130z that moves the array plate 101 relative to a base material (not shown) in a Z direction perpendicular to the plane defined by the X and Y directions. The z-direction scanning means 130z includes a linear stage 112z that moves the linear stage 109 in the Z direction, a pulse motor 110 that drives the linear stage 112z, and a motor driver 111 that controls the pulse motor 110. The scanning means 130 is fixed to a scanning means base material (housing) (not shown). Note that although a pulse motor or the like is exemplified here as a drive source that generates a drive force, other known drive mechanisms such as motors may also be used as the drive source.

[0021] The piston crank mechanism 120 reciprocates the light projecting unit 104 in the widthwise direction of the array plate 101. Due to the reciprocating motion of the light projecting unit 104, the excitation light from the light source 102 is scanned in the widthwise direction of the array plate 101. Note that the widthwise direction of the array plate 101 is referred to as the main scanning direction, and the reciprocating scanning by the piston crank mechanism 120 is referred to as main scanning. The main scanning stroke shown here is 30 mm. The light projecting unit 104 is limited in its operating direction to the main scanning direction by a guide (not shown) in the main scanning direction. The pulse motor 107 is a pulse motor for rotating the piston crank mechanism 120 at high speed, and the rotation speed of the pulse motor 107 shown here is in the range of 100 to 10,000 rpm, preferably 500 to 2,000 rpm.

[0022] <Position Acquisition Means> The specimen measurement device 1 includes a position acquisition means 138 (see FIG. 3) that acquires position information related to the irradiation position of the primary light on the array plate 101. The position acquisition means 138 includes a linear encoder 108 that moves integrally with the light projecting unit 104 shown in FIG. 1, and a coordinate calculation circuit 310, which will be described later. The linear encoder 108 is a linear encoder installed on the piston crank mechanism 120, and measures the position of the light projecting unit 104 in the main scanning direction. The linear encoder 108 outputs a phase difference pulse voltage consisting of A phase, B phase, and Z phase according to the position in the main scanning direction.

[0023] Here, with reference to FIG. 16 , a specific configuration of the piston crank mechanism 120 used in the specimen measurement device 1 shown in FIG. 1 will be described. FIG. 16 shows an example of a schematic configuration of the piston crank mechanism 120. The piston crank mechanism 120 includes a piston crank unit 123 including a crank 118 and a connecting rod 119, and a rotation drive unit 121. FIG. 16 shows the positional relationship between the crank 118, the connecting rod 119, and the light projector 104 in the piston crank mechanism 120. The crank 118 is a crank that constitutes the piston crank mechanism 120, and is connected to the rotation shaft of the pulse motor 107 and the connecting rod 119 via a joint. In this embodiment, the pulse motor 107 constitutes the rotation drive unit 121 in the present disclosure and is connected to one end of the crank 118. The other end of the crank 118 is connected to one end of the connecting rod 119. The other end of the connecting rod 119 is connected to the light projector 104. With this configuration, the rotational movement of the rotary drive unit 121 enables the light projecting unit 104 to perform a one-dimensional reciprocating movement along the direction of the arrow shown in FIG.

[0024] The linear stage 109 moves the array plate 101 in a direction perpendicular to the main scanning direction within a horizontal plane. The linear stage 109 is composed of a ball screw, an origin sensor, and the like. Hereinafter, scanning in a direction perpendicular to the main scanning direction within a horizontal plane will be referred to as sub-scanning. A mounting table for the array plate 101 is provided on the linear stage 109, and the user places the array plate 101 to be measured on the mounting table in advance. A pulse motor 110 is connected to the linear stage 109. The rotational motion of the pulse motor 110 is converted into linear motion by the ball screw of the linear stage 109. A motor driver 111 rotates the pulse motor 110. When the motor driver 111 inputs one pulse signal to the pulse motor 110, the pulse motor 110 rotates 0.72°, and the array plate 101 moves 2 μm in the sub-scanning direction.

[0025] The linear stage 112z moves the array plate 101 in the vertical direction. The linear stage 112z is composed of a ball screw, an origin sensor, and the like. Hereinafter, vertical scanning will be referred to as height scanning. The pulse motor 113 is connected to the linear stage 112z. The rotational motion of the pulse motor 113 is converted into linear motion by the ball screw of the linear stage 112z. The motor driver 114 rotates the pulse motor 113. When the motor driver 114 inputs a signal of one pulse to the pulse motor 113, the pulse motor 113 rotates 0.72°, and the array plate 101 moves 1 μm upward in the vertical direction (height scanning). The motor driver 115 rotates the pulse motor 107. When the motor driver 115 inputs a signal of one pulse to the pulse motor 107, the pulse motor 107 rotates 0.72°, and the light projector 104 moves along the main scanning direction. Note that the configuration described here regarding the relative movement of the light projecting unit 104 (the irradiation position of the excitation light) relative to the array plate 101 is just one example, and as long as relative movement between these configurations is possible, there is no limitation as to which of these configurations moves and how.

[0026] The analyte measurement device 1 further includes a controller circuit 116. The controller circuit 116 controls the entire analyte measurement device 1 and is composed of an FPGA, a CPU, memory, embedded software, and the like. More specifically, the controller circuit 116 controls the light source 102, the motor driver 111, the motor driver 114, and the motor driver 115 to scan the excitation light in the main scanning direction, the sub-scanning direction, and the height direction on the array plate 101. The controller circuit 116 also acquires information regarding the irradiation position Pi of the irradiation light (primary light) output from the linear encoder 108 and optical signal data from the optical sensor 105. The controller circuit 116 then stores this information in a first storage unit 171 and a second storage unit 172 (see FIG. 17 ), which will be described later. The first storage unit 171 stores the information regarding the irradiation position Pi in a readable state, thereby storing the information regarding the irradiation position Pi (at least temporarily) for a period during which it can be used by a determination unit 173 (see FIG. 17 ), which will be described later.

[0027] The controller circuit 116 includes a means for acquiring height information and tilt information of the array plate 101 and a means for generating pulse trains for main scanning, sub-scanning, and height scanning. These means correspond to the motor control circuits 307-309 and the coordinate calculation circuit 310 shown in FIG. 3 , which will be described later. As shown in FIG. 3 , the controller circuit 116 also includes an AD converter control circuit 306 as AD converter control means and a synchronization circuit 311 as synchronization control means. The controller circuit 116 also includes a data acquisition control circuit 312 as data acquisition control means and a memory 303 as storage means. In other words, in the embodiment described below, these means provided in the controller circuit 116 are configured by the various circuits shown in FIG. 3 . In this case, the AD converter control means issues a conversion instruction to the AD converter 705 (see FIG. 7 ) of the receiving circuit 106, which performs AD conversion on the electrical signal corresponding to the light intensity of the optical sensor 105, thereby acquiring optical signal data as a digital value. In this case, in the following embodiment, the controller circuit 116 uses these configurations to acquire optical signal data at a cycle shorter than the shortest time it takes for the irradiation position Pi of the primary light to pass through the interval between the target positions Pt.

[0028] The synchronization circuit 311, which serves as a synchronization control means, controls the sub-scanning and height scanning in synchronization with the position information Iip of the primary light irradiation position Pi. This corrects the thickness and tilt of the array plate 101 when the light projector 104 corresponding to the primary light irradiation position Pi is outside the imaging range on the array plate 101, and focuses the excitation light on the entire surface of the array plate 101 where the spot is fixed. Furthermore, the synchronization circuit 311 controls the timing of light emission from the light source 102 and the transfer of optical signal data to memory. As part of the synchronization control, the synchronization circuit 311 instructs the data acquisition control means to transfer optical signal data to the memory 303 when the primary light irradiation position Pi reaches the target position Pt so that the excitation light can be irradiated to the predetermined position. Furthermore, the synchronization circuit 311 controls the start of the sub-scanning and height scanning, and the turning on and off of the semiconductor lasers of each wavelength of the light source 102 when the light projector 104 reaches a position where the excitation light can be irradiated to a predetermined position near the edge of the array plate 101.

[0029] <First Holding Means> Here, details of the synchronization circuit 311 provided in the specimen measurement device 1 in this embodiment will be described with reference to FIG. 17. FIG. 17 is a block diagram showing the functional configuration provided in the synchronization circuit 311 in this embodiment. In this embodiment, the synchronization circuit 311 includes a first holding means 171, a second holding means 172, and a determination means 173. The first holding means 171 shown in FIG. 17 holds data Itp relating to the target position Pt with respect to the irradiation position Pi. Note that, although an example has been given here in which the first holding means 171 is arranged in the synchronization circuit 311, the arrangement of the first holding means 171 is not limited to this example.

[0030] 17 , the synchronization circuit 311 in this embodiment also includes second holding means 172 that holds digital data converted from the detection signal by the conversion means 146. Note that, although the configuration in which the second holding means 172 is disposed in the synchronization circuit 311 has been exemplified here, the arrangement of the second holding means 172 is not limited to this example.

[0031] 17 , the synchronization circuit 311 in this embodiment includes a determination unit 173 that determines whether the irradiation position Pi has reached the target position Pt. The determination unit 173 determines whether the irradiation position Pi of the primary light has reached the target position Pt, based on information about the relative position obtained from the linear encoder 108 or the like. Note that, although the configuration in which the determination unit 173 is disposed in the synchronization circuit 311 has been exemplified here, the arrangement of the second holding unit 172 is not limited to this example.

[0032] <Transfer Means> As shown in FIG. 3, the specimen measurement device 1 also includes a data acquisition control circuit 312 (see FIG. 3) that selects data corresponding to the irradiation position corresponding to the target position from the digital data stored in the second storage means 172 in accordance with the determination result by the determination means 173. In this embodiment, the memory control circuit 304 connected to the data acquisition control circuit 312 functions as a transfer means that transfers the selected data to the storage means (memory 303). The data acquisition control circuit 312 temporarily stores optical signal data from the AD converter control means, and the memory control circuit 304 transfers the latest optical signal data to the memory based on a synchronization control instruction from the synchronization circuit 311. In the following embodiment, if new optical signal data is acquired without an instruction from the synchronization circuit 311, the old optical signal data is overwritten. As a result, optical signal data is acquired at a regular interval, but the optical signal data acquired closest to the time when the primary light irradiation position Pi reaches the target position Pt is selectively transferred to and stored in the memory 303.

[0033] The specimen measurement device 1 shown in Figure 3 further includes a user interface 117. The user interface 117 is used to receive instructions from the user and display results, and is composed of a keyboard, mouse, and display. The controller circuit 116 receives instructions to capture an image of the array plate 101 and presents image data based on the optical signal data to the user via the user interface 117. When capturing an image of the array plate 101, the user also specifies the capture range and pixel pitch in the main scanning direction and sub-scanning direction via the GUI on the user interface 117.

[0034] Next, the specimens targeted by the present disclosure will be described using FIGS. 2A and 2B. FIG. 2A is a top view of the array plate 101, and FIG. 2B is a side view of the array plate 101. The array plate 101 includes a glass slide 201 as a substrate and spots 202 provided on the glass slide 201. A biological substance containing a peptide bond is immobilized on each spot 202. Multiple spots 202 are provided on the glass slide 201, and one type of biological substance is immobilized on each spot. In the following embodiment, the diameter of the spots is approximately 100 μm, and the spot spacing (the distance from the end of one spot to the end of the next spot) is 200 μm. The array plate has a short-side length of 25 mm and a long-side length of 75 mm.

[0035] Here, the origin is point 203 at the upper left (corner) of the array plate 101, the rightward direction in the short direction is the positive X-axis direction, and the downward direction in the long direction is the Y-axis direction. If the units of X and Y coordinates are μm, the coordinates of the four corners of the array plate are (0,0), (25000,0), (0,75000), and (25000,75000), respectively. If the stroke of the piston crank mechanism 120 is 30 mm, this stroke is 5 mm longer than the length of the array plate 101. In other words, in the main scan, the excitation light scans a range 2.5 mm longer on both sides of the array plate, and the X coordinate of the scanned range is in the range of -2500 to 27500.

[0036] Meanwhile, on the glass slide 201, the spot 202 is provided within the range of an area 204. That is, depending on the creation of the spot 202 and the user's gripping of the glass slide 201, there are areas on the glass slide 201 where the spot 202 exists and areas where it does not. In the following embodiment, the coordinates of the four corners of the area 204 are (2000, 2000), (23000, 2000), (2000, 65000), and (23000, 65000), respectively. The range of the Y coordinate in the sub-scanning direction can be specified by the user.

[0037] 3, which was partially mentioned above, is a block diagram showing the internal configuration of the controller circuit 116 according to the first embodiment of the present disclosure. The controller circuit 116 has a CPU 301 for running control software that controls the entire specimen measurement device 1, and the CPU 301 is composed of a microprocessor, cache memory, etc. The CPU 301 is also connected to various peripheral circuits, which will be described later, via a bus 302. The CPU 301 may also include an image processing unit 301a, which uses digital data based on optical signal data stored in a memory 303, which will be described later, to image the characteristics of the specimen.

[0038] The various peripheral circuits include a memory 303, a memory control circuit 304, a light source control circuit 305, an AD converter control circuit 306, motor control circuits 307 to 309, a coordinate calculation circuit 310, and a synchronization circuit 311. The various peripheral circuits also include a data acquisition control circuit 312, a communication circuit 313, and a UI (User Interface) control circuit 314.

[0039] <Storage Unit> The memory 303, which can be referred to as a storage means, stores pixel values ​​and coordinate value data constituting a fluorescent image. The memory 303 may also store imaging conditions and control parameters of the specimen measurement device 1 input by the user, as well as optical signal data. The memory 303 is composed of a DDR (Double Data Rate), a 4-SDRAM (Synchronous Dynamic Random Access Memory), an SSD (Solid State Drive), or the like. The memory control circuit 304 controls the memory 303 based on access commands to the memory 303 received via the bus 302. The light source control circuit 305 is a light source control circuit used by the CPU 301 to control the light source 102, and is composed of an interface conversion circuit, a DA converter, and the like. The CPU 301 can control the on / off of laser irradiation from the light source 102 and the amount of excitation light irradiated via the light source control circuit 305. In other words, the light source control circuit 305 is an element of the light irradiation means 122 configured to turn on and off the light emission of the light source 102 .

[0040] The AD converter control circuit 306 controls the AD converter of the receiving circuit 106 described above based on instructions from the CPU 301, and acquires optical signal data. The AD converter control circuit 306 outputs a conversion start signal to the AD converter of the receiving circuit 106. The AD converter samples the analog signal in the receiving circuit at the timing when the conversion start signal is input, converts this into optical signal data, and outputs it to the AD converter control circuit 306. The AD converter control circuit 306 transfers the optical signal data input from the AD converter to the data acquisition control circuit 312.

[0041] The motor control circuit 307 generates a control signal for the motor driver 115 for the main scanning motor based on instructions from the CPU 301. When the CPU 301 instructs the rotation speed, acceleration, movement amount, rotation direction, and rotation start timing of the pulse motor 107, the motor control circuit 307 generates a drive pulse voltage corresponding to the instructions. The motor control circuit 308 generates a control signal for the motor driver 111 for the sub-scanning motor based on instructions from the CPU 301. When the CPU 301 instructs the rotation speed, acceleration, movement amount, rotation direction, and rotation start timing of the pulse motor 110, the motor control circuit 308 generates a drive pulse voltage corresponding to the instructions. The motor control circuit 309 generates a control signal for the motor driver 114 for the height scanning motor based on instructions from the CPU 301. When the CPU 301 instructs the rotation speed, acceleration, movement amount, rotation direction, and rotation start timing of the pulse motor 113, the motor control circuit 309 generates a drive pulse voltage corresponding to the instructions.

[0042] The coordinate calculation circuit 310 counts the two phase difference pulse signals of A and B phases from the linear encoder 108 and calculates the position of the light-projecting unit 104. Note that the resolution of the linear encoder 108 is assumed to be 1 μm here. When the level of the A-phase signal or B-phase signal changes and the A-phase signal is ahead in phase of the B-phase signal, the coordinate calculation circuit 310 increases the coordinate of the light-projecting unit 104 by 1 μm. When the level of the A-phase signal or B-phase signal changes and the B-phase signal is ahead in phase of the A-phase signal, the coordinate calculation circuit 310 decreases the coordinate of the light-projecting unit 104 by 1 μm.

[0043] The synchronization circuit 311 generates trigger signals to be sent to the data acquisition control circuit 312, motor control circuit 308, motor control circuit 309, and light source control circuit 305 based on the coordinate information obtained from the coordinate calculation circuit 310. Hereinafter, the trigger signal to the data acquisition control circuit 312 will be referred to as a data acquisition trigger signal. The trigger signal to the motor control circuit 308 will be referred to as a sub-scan trigger signal. The trigger signal to the motor control circuit 309 will be referred to as a height scanning trigger signal. In addition, the trigger signal to the light source control circuit 305 will be referred to as a wavelength switching trigger signal.

[0044] When the motor control circuit 308 receives the sub-scan trigger signal, it outputs a drive pulse train corresponding to the amount of movement equal to the pixel pitch in the sub-scan direction to the motor driver 111. The pixel pitch in the sub-scan direction is specified by the user when shooting starts and is stored in the memory 303. When the motor control circuit 309 receives the height scan trigger signal, it outputs a drive pulse train corresponding to the amount of movement in the height scan direction to the motor driver 114. The amount of movement in the height scan direction is calculated by the CPU 301. The calculation method will be described later.

[0045] The data acquisition control circuit 312 transfers the optical signal data output from the AD converter control circuit 306 to the memory 303 via the memory control circuit 304. In this embodiment, the data acquisition control circuit 312 is composed of a buffer memory, a DMA controller, etc. When the data acquisition control circuit 312 receives a data acquisition trigger signal from the synchronization circuit 311, it transfers the latest optical signal data stored in the internal buffer memory to the memory 303.

[0046] The communication circuit 313 connects the specimen measurement device 1 to an external network. In this embodiment, the communication circuit 313 uses a communication protocol conforming to the Ethernet (registered trademark) standard as a communication method. By connecting the specimen measurement device 1 to an external PC or server, the communication circuit 313 makes it possible to remotely control specimen imaging and store data in external large-capacity storage.

[0047] The UI control circuit 314 connects the specimen measurement device 1 to the user interface 117. The UI control circuit 314 is composed of an input circuit for inputting data from a keyboard or mouse, and an image display circuit for controlling the display. The user interface 117 can also be configured as a monitor, in which case it displays images to the user on the display screen and accepts operation instructions from the user.

[0048] In this embodiment, the peripheral circuits constituting the controller circuit 116 are implemented on a semiconductor chip such as an FPGA or an ASIC. These peripheral circuits operate in synchronization with a clock. In this embodiment, the clock frequency is 1.00 MHz.

[0049] Next, the imaging operation of the array plate 101 in this embodiment will be described using Figures 4, 5A, 5B, 6A, and 6B. Figure 4 shows the operational flow when imaging the array plate 101 using the specimen measurement device 1 according to this embodiment. Figures 5A and 5B are diagrams illustrating the manner of sub-scanning on the array plate 101 during imaging, with Figure 5A being a top view of the array plate 101 and Figure 5B being a schematic diagram illustrating the trajectory of excitation light on the array plate 101. Figures 6A and 6B are diagrams illustrating the positional relationship of vertical scanning of the array plate 101 during imaging, with Figures 6A and 6B being diagrams illustrating an example of the array plate 101 viewed from the side. Note that the thickness and inclination of the array plate 101 installed in the specimen measurement device 1 differ between Figures 6A and 6B.

[0050] When the user inputs an instruction to start imaging the array plate 101 via the user interface 117, the processing from step S401 onwards is started. In step S401, the CPU 301 reads the imaging conditions specified by the user via the user interface 117 and stores them in the memory 303. The CPU 301 also sets information based on the imaging conditions in various circuits within the controller circuit 116.

[0051] As imaging conditions, points 501 (X1, Y1) and 502 (X2, Y2) indicating the imaging range on the array plate 101, pixel pitch Xp in the main scanning direction, pixel pitch Yp in the sub-scanning direction, and rotation speed Xs in the main scanning direction are input. In this embodiment, the following are input: X1 = 500, X2 = 22,500, Y1 = 500, Y2 = 64,500, Xp = 2 μm, Yp = 2 μm, and Xs = 1,200 rpm. Here, a rectangular imaging area 503 on the array plate 101 with points 501 and 502 as diagonals is referred to as the imaging area. Furthermore, the number of pixels in the main scanning direction Nx = (X2 - X1) / Xp and the number of pixels in the sub-scanning direction Ny = (Y2 - Y1) / Yp are calculated. In this embodiment, according to the above settings, Nx = 11,000 and Ny = 32,000. After the reading of the photographing conditions and the above settings are completed, the flow proceeds to step S402.

[0052] In step S402, the CPU 301 acquires height information and tilt information of the array plate 101. Here, the amount of movement of the linear stage 112 required to focus the excitation light on the upper surface of the slide glass 201 constituting the array plate 101, i.e., the surface on which the spot 202 is located, is referred to as height information. Also, the tilt of the array plate 101 in the sub-scanning direction is referred to as tilt information. In step S402, Z coordinates Z3 and Z4, which are height information relative to two Y coordinates Y3 and Y4, are acquired by measurement.

[0053] In this embodiment, it is assumed that Y3 = 750 and Y4 = 65,000 are input. Here, if the tilt is as shown in FIG. 6A, Z4 > Z3, and if the tilt is as shown in FIG. 6B, Z3 < Z4. The method for obtaining height information will be described later. Furthermore, tilt information K is calculated using the following formula 1: K = (Z4 - Z3) / (Y4 - Y3) (Formula 1) Once the tilt is obtained, the flow proceeds to step S403.

[0054] In step S403, the CPU 301 calculates the target height of each row of spots according to the imaging conditions and the height information and tilt information K. The target height Z(Y) at the coordinate Y of an arbitrary sub-scanning position is given by the following equation 2. Note that the target height 601 corresponds to the Z coordinate of the surface of the array plate 101 in Figures 6A and 6B. Z(Y) = K x (Y - Y3) + Z3 (Equation 2) Once the target height is calculated, the flow proceeds to step S404.

[0055] In step S404, the CPU 301 issues instructions to the motor control circuits 307, 308, and 309 to move the array plate 101 and the light projector 104 to the imaging start position. In this embodiment, the X coordinate of the imaging start position is the end of the scanning range of the piston crank mechanism 120, and the X coordinate value is -2500. The Y coordinate of the imaging start position is Y1 specified in the imaging conditions. The Z coordinate Z1 of the imaging start position is expressed as Z(Y1) in equation 1. Once the array plate 101 and the light projector 104 have been moved to the imaging start position, the flow proceeds to step S405.

[0056] In step S405, the CPU 301 issues an instruction to the motor control circuit 307 to rotate the pulse motor 107 at a rotational speed Xs. This causes the light projector 104 to start reciprocating in the X direction. At this time, the linear encoder 108 and the coordinate calculation circuit 310 calculate the X coordinate of the light projector 104 for each clock, and the calculation result is output to the synchronization circuit 311. In this embodiment, information regarding the calculated relative position of the light projector 104 with respect to the array plate 101 is temporarily stored in a buffer memory provided in the synchronization circuit, and is transmitted to and stored in the memory 303 as necessary. Once the reciprocating motion has started, the flow proceeds to step S406.

[0057] In step S406, the CPU 301 issues an instruction to the light source control circuit 305 to cause the light source 102 to start emitting light. This initiates light irradiation (irradiation of excitation light, hereinafter referred to as light irradiation) onto the array plate 101 via the light projector 104. At this time, the synchronization circuit 311 is used to irradiate the array plate 101 with light of two wavelengths, 670 nm and 780 nm, while alternately switching between forward and backward main scanning. In this manner, in this embodiment, the optical sensor 105 does not simultaneously receive the fluorescence caused by the two wavelengths of light, but rather receives the fluorescence caused by the light of each wavelength individually. This reduces crosstalk between wavelengths that can occur when light is received simultaneously, improving the measurement accuracy of the optical signal data. Once light irradiation has begun, the flow proceeds to step S407.

[0058] In step S407, the synchronization circuit 311 determines whether the irradiation position Pi of the primary light on the array plate 101 has reached the wavelength switching position. Specifically, the synchronization circuit 311 determines that the wavelength switching position has been reached when the X coordinate of the irradiation position of light irradiated from the light projector 104 on the array plate 101, output from the coordinate calculation circuit 310, moves to predetermined positions X3 and X4 outside the imaging area. Here, this light irradiation position is calculated as the relative position of the light projector 104 in the XY plane on the array plate 101. Note that in this embodiment, the coordinates of positions X3 and X4 are set in advance and stored in the synchronization circuit 311. Furthermore, positions X3 and X4, which are the wavelength switching positions, are set to positions closer to the stroke ends of the light projector 104 than positions X1 and X2, which are the line feed positions. This is because switching the wavelength requires a certain amount of time, and the wavelength switching must be completed while the light projector 104 is outside the imaging area. In this embodiment, position X3=200 and position X4=22800 are adopted.

[0059] If the synchronization circuit 311 determines that the light irradiation position has reached the wavelength switching position, the synchronization circuit 311 outputs a wavelength switching trigger signal to the light source control circuit 305, and the flow proceeds to step S421. On the other hand, if the synchronization circuit 311 determines that the wavelength switching position has not been reached, the synchronization circuit 311 does not output a wavelength switching trigger signal, and the flow proceeds to step S408.

[0060] In step S408, the synchronization circuit 311 determines whether the irradiation position Pi of the primary light has reached the line feed position. The synchronization circuit 311 determines that the line feed position has been reached when the X coordinate of the light projector 104 output from the coordinate calculation circuit 310 moves from inside the shooting area to outside the shooting area. In this embodiment, if the current main scanning direction is the backward direction (the direction in which the X coordinate decreases), the synchronization circuit 311 determines that the line feed position has been reached when the X coordinate of the light projector 104 becomes smaller than X1. Furthermore, in this embodiment, the synchronization circuit 311 does not determine that the line feed position has been reached when the X coordinate of the light projector 104 becomes larger than X2.

[0061] The forward scan is represented by trajectory 504 in Fig. 5B. The backward scan is represented by trajectory 506 in Fig. 5B, which follows the same trajectory as the forward scan, but in the opposite direction, to return to the line feed position. The initial value of the main scanning direction is the forward scan, and thereafter, each time the stroke end is reached, the scanning direction is alternately changed between the backward scan and the forward scan.

[0062] If the synchronization circuit 311 determines that the line feed position has been reached, the synchronization circuit 311 outputs a sub-scan trigger signal and a height scan trigger signal, and executes the processes of steps S417 and S418 in parallel. If the synchronization circuit 311 determines that the line feed position has not been reached, the synchronization circuit 311 does not output the sub-scan trigger signal and the height scan trigger signal, and the flow proceeds to step S409.

[0063] In step S409, the determination means 173 included in the synchronization circuit 311 determines whether the irradiation position Pi of the primary light has reached the target position Pt. Point 507 in FIG. 5B indicates the position of a pixel to be imaged on the array plate 101. In this embodiment, the target position Pt is a point on the array plate 101 where optical signal data is acquired to generate a pixel. The X-coordinate P(N) of the Nth target position Pt is expressed by the following equation 3: P(N) = X1 + Xp × N (N = 0, 1, ... Nx - 1) (Equation 3). The target positions are multiple points on the trajectory, with a pitch of Xp in the X direction and a pitch of Yp in the Y direction. In this embodiment, these points coincide with the pixel position, point 507. The initial value of the target position Pt is P(0), which is stored in a buffer memory or the like provided within the synchronization circuit 311. In the initial determination of the target position Pt, it is determined that the target position Pt has been reached when the X coordinate of the light-projecting unit 104 output from the coordinate calculation circuit 310 passes P(0) in the forward direction (the direction in which the X coordinate increases). In the second and subsequent determinations of the target position Pt, it is determined that the target position Pt has been reached when the X coordinate of the light-projecting unit 104 output from the coordinate calculation circuit 310 passes the target position Pt for which the irradiation position Pi has been updated in step S411, which will be described later. In this embodiment, the target position is generated each time based on the pixel pitch used in image generation and the position information Iip of the irradiation position Pi from the linear encoder 108, which is obtained by the synchronization circuit 311. Information regarding the target position Pt is stored each time in a buffer memory provided in the synchronization circuit 311. Information regarding the target position Pt can also be transferred to and stored in the memory 303 as necessary. That is, in this embodiment, the synchronization circuit 311 temporarily holds a target position Pt calculated each time from the pixel pitch and the relative position of the light-projecting unit 104 with respect to the array plate 101. Then, the synchronization circuit 311 determines whether the irradiation position Pi of the primary light has reached the target position Pt based on the held target position and position information obtained from the encoder.

[0064] If the synchronization circuit 311 determines that the target position Pt has been reached, the synchronization circuit 311 outputs a data acquisition trigger signal, and the flow proceeds to step S410. If the synchronization circuit 311 determines that the target position Pt has not yet been reached, the synchronization circuit 311 does not output a data acquisition trigger signal, and the flow proceeds to step S412.

[0065] In step S410, the AD converter control circuit 306 outputs a conversion start signal, and the output voltage from the optical sensor 105 is amplified by the receiving circuit 106, filtered, and then AD converted. In this embodiment, the conversion start signal is a signal with a constant period, which is longer than the conversion time of the AD converter but shorter than the time it takes for the light projecting unit 104 to move through the pixel pitch Xp. The AD-converted optical signal data is temporarily stored in a buffer memory inside the data acquisition control circuit 312. The buffer memory can store one optical signal data per AD converter, and the latest optical signal data is overwritten and saved each time AD conversion is performed. In this embodiment, an AD converter corresponding to the wavelength of each light source is provided, and optical signal data reflecting the amount of fluorescent light excited by the wavelength of each light source is saved.

[0066] When the data acquisition control circuit 312 receives a data acquisition trigger signal, it reads out the optical signal data stored in the buffer memory and outputs it to the memory control circuit 304 via an internal DMA controller and the bus 302. The memory control circuit 304 stores the optical signal data at a specified address in the memory 303. The address in the memory 303 is incremented each time a data acquisition trigger signal is received.

[0067] After storing Nx×Ny pieces of optical signal data corresponding to the wavelengths of each light source in memory 303, the data acquisition control circuit 312 sets an internal data acquisition completion register to 1. When Nx×Ny pieces of optical signal data corresponding to the wavelengths of each light source have not been stored in memory 303, the data acquisition control circuit 312 sets an internal data acquisition completion register to 0. Once the optical signal data at target position Pt has been stored, the flow proceeds to step S411.

[0068] Next, in step S411, the first holding means 171 (synchronization circuit 311) updates the target position Pt stored therein. If the current main scanning direction is the forward direction (the direction in which the X coordinate increases), the target position Pt(N) is incremented and updated to P(N+1). If the current main scanning direction is the backward direction (the direction in which the X coordinate decreases), the target position Pt(N) is updated to P(N-1). After the target position Pt has been updated, the flow proceeds to step S412.

[0069] In step S412, the CPU 301 determines whether the acquisition of optical signal data has finished. The CPU 301 reads the data acquisition completion register of the data acquisition control circuit 312, and if the value is 1, it determines that the acquisition of optical signal data has finished, and moves the flow to step S413. On the other hand, if the value is 0, it determines that the acquisition of optical signal data has not finished, and moves the flow to step S407.

[0070] In step S413, the CPU 301 issues an instruction to the light source control circuit 305 to stop the light emission of the light source 102. This stops the light irradiation onto the array plate 101 via the light projector 104. After the light irradiation has stopped, the flow proceeds to step S414.

[0071] In step S414, the CPU 301 issues an instruction to the motor control circuit 307 to stop the rotation of the pulse motor 107. This stops the reciprocating motion of the light projector 104 in the X direction. After the reciprocating motion has stopped, the flow proceeds to step S415, where the CPU 301 issues an instruction to the motor control circuits 307, 308, and 309 to move the array plate 101 and the light projector 104 to a stop position. The X, Y, and Z coordinates of the stop position are 0. Movement to the stop position is achieved by returning each axis to its origin using the Z-phase pulse signal of the linear encoder 108, the origin sensor signal in the linear stage 109, and the origin sensor signal of the linear stage 112. Once the array plate 101 and the light projector 104 have returned to their origins for each axis, the flow proceeds to step S416.

[0072] In step S416, the CPU 301 reads out the Nx x Ny pieces of optical signal data corresponding to the wavelengths of the respective light sources from the memory 303, performs data compression and format conversion, and creates a captured image file in TIFF format. The captured image file is saved in the memory 303 and presented to the user via the UI control circuit 314 and the user interface 117. In addition, the captured image file can be transferred to an external data server via the communication circuit 313 in response to an instruction from the user.

[0073] In step S417, the CPU 301 issues an instruction to the motor control circuit 308 to move the array plate 101 by Yp in the sub-scanning direction. If the amount of movement of the array plate 101 when one pulse of a voltage pulse signal is sent to the motor driver 111 is My, the number of pulses output by the motor control circuit 308 to the motor driver 111 is Yp / My. In this embodiment, My = 2 μm. The trajectory of the light irradiation position during sub-scanning is represented by trajectory 505 in FIG. 5B, and the movement distance in the Y direction at that time is Yp. Based on the instruction from the CPU 301, the motor control circuit 308 outputs a pulse signal to the motor driver 111 at a speed such that the sub-scanning is completed while the light projector 104 is outside the imaging area in the main scanning direction.

[0074] In step S418, the CPU 301 reads the target heights before and after the sub-scanning from the memory 303. That is, it reads Z(Y+Yp) and Z(Y) and calculates the amount of movement for the height scanning. Although the amount of movement for the height scanning differs depending on the current Y coordinate, it calculates the number of output pulses and the direction of rotation of the motor so that the height in the Z direction after the movement is closest to the target height Z(Y+Yp) for the Y coordinate after the sub-scanning in step S417.

[0075] The calculation direction is described below. The amount of movement of the array plate 101 when one pulse of a voltage pulse signal is sent to the motor driver 114 is defined as Mz. In this embodiment, Mz = 1 μm. The multiple of Mz closest to a certain number x is defined as RoundMz(x), the absolute value of the certain number is defined as ABS(x), and the sign of the certain number is defined as Sign(x). The multiple of Mz closest to the target height is called the target pulse number. The target pulse number takes a discrete value and corresponds to the pulse number 602 at each spot position in FIGS. 6A and 6B. Here, the number of pulses output by the motor control circuit 309 to the motor driver 114 is expressed by the following equation 4. Zp=ABS(RoundMz(Z(Y+Yp))−RoundMz(Z(Y))) (Equation 4) In Equation 4, RoundMz(Z(Y)) becomes a discrete value close to the surface of the array plate 101, as shown by the pulse number 602 in Figures 6A and 6B.

[0076] The tilt direction Dir of the array plate 101 in the height direction is expressed by the following equation 5: Dir=Sign(RoundMz(Z(Y+Yp))-RoundMz(Z(Y))) (Equation 5) In equation 5, the positive direction of Dir is defined as vertically upward, and as the direction in which the distance between the light projecting unit 104 and the array plate 101 increases. When the array plate 101 is tilted in the manner shown in FIG. 6A, the value of Dir is 1, and when it is tilted in the manner shown in FIG. 6B, the value of Dir is -1. Once the target height is read, the flow proceeds to step S419.

[0077] In step S419, the CPU 301 issues an instruction to the motor control circuit 309 to move the array plate 101 by Zp in a direction that cancels out the tilt direction Dir. When the value of Dir is positive, the array plate is moved downward, and when the value of Dir is negative, the array plate is moved upward. At this time, the number of pulses output by the motor control circuit 309 to the motor driver 111 is Zp / Mz. Based on the instruction from the CPU 301, the motor control circuit 309 outputs a pulse signal to the motor driver 114 at a speed that completes the height scan while the light projecting unit 104 is outside the imaging area in the main scanning direction. When the sub-scanning in step S417 and the height scan in step S419 are completed, the flow proceeds to step S420.

[0078] In step S420, the synchronization circuit 311 increments the current Y coordinate by Yp from the previous Y coordinate, and the flow proceeds to step S412.

[0079] In step S421, the light source control circuit 305 switches between emitting and stopping the 670 nm and 780 nm semiconductor lasers based on information about the wavelength switching trigger signal from the synchronization circuit 311. More specifically, if the current main scanning direction is the forward direction (the direction in which the X coordinate increases), the 780 nm laser is emitted when the X coordinate of the light projector 104 passes position X3. Furthermore, the 780 nm laser is stopped when it passes position X4. If the main scanning direction is the forward direction (the direction in which the X coordinate increases), the 670 nm laser is stopped. If the current main scanning direction is the backward direction (the direction in which the X coordinate decreases), the 670 nm laser is emitted when the X coordinate of the light projector 104 becomes smaller than position X4. Furthermore, the 670 nm laser is stopped when it becomes smaller than position X3. If the main scanning direction is the forward direction (the direction in which the X coordinate increases), the 780 nm laser is stopped. When the switching of the irradiated light is completed, the flow proceeds to step S412.

[0080] By executing the above process, irradiation of each spot on the array plate 101 with laser light of two wavelengths and imaging of the spot based on the fluorescence caused by the laser light are completed.

[0081] Next, the optical sensor 105 and receiving circuit 106 in the specimen measurement device 1 according to the present embodiment will be described with reference to the drawings. Fig. 7 is a functional block diagram showing details of the optical sensor 105 and receiving circuit 106 in the specimen measurement device 1 according to the present embodiment.

[0082] The optical sensor 105 used in this embodiment includes photomultiplier tubes 701 and 710 and photodiodes 709 and 711. The photomultiplier tube 701 is excited by 780 nm light, converts the amount of fluorescence emitted from the specimen into a current, and outputs this. The photodiode 709 converts the amount of 780 nm light reflected by the array plate 101 into a current, and outputs this. The photomultiplier tube 710 is excited by 670 nm light, converts the amount of fluorescence emitted from the specimen into a current, and outputs this. The photodiode 711 converts the amount of 670 nm light reflected by the array plate 101 into a current, and outputs this.

[0083] The receiving circuit 106 includes a current-voltage (IV) amplifier 702, a level conversion circuit 703, a filter circuit 704, and an AD converter (ADC) 705. The current-voltage conversion amplifier 702 outputs a voltage proportional to the current from the photomultiplier tube 701. In this embodiment, the output voltage range of the current-voltage conversion amplifier is −10 V to 10 V. The level conversion circuit 703 is a circuit that halves the level of the output voltage from the current-voltage conversion amplifier 702 and is composed of a resistor network and a clamp circuit. The level conversion circuit clamps negative voltages to 0 V, and the voltage level of positive voltages is halved by a voltage divider circuit. In this embodiment, because the direction of the current from the photomultiplier tube 701 causes the voltage of the fluorescence signal to be approximately 0 V or higher, the level conversion circuit 703 is provided to limit the range of voltages converted into digital values ​​by the downstream AD converter 705 to positive voltages, thereby increasing the resolution of the digital values. As a result, the output voltage range after level conversion is 0 V to 5 V. The filter circuit 704 cuts out high-frequency signals of 500 kHz or higher from the output voltage of the level conversion circuit 703. The provision of the filter circuit 704 eliminates aliasing that occurs during sampling. The AD converter 705 is a 16-bit AD converter that converts the output from the filter circuit 704 into a digital value ranging from 0 to 65535.

[0084] The AD converter control circuit 306 and the AD converter 705 exchange control signals 706, 707, and 708. The control signal 706 is a conversion start signal that starts sampling and AD conversion. The control signal 707 is a clock signal used when transferring optical signal data from the AD converter 705 to the AD converter control circuit 306. In this embodiment, the clock frequency of the clock signal 707 is 50 MHz. The control signal 708 is a data signal used when transferring optical signal data from the AD converter 705 to the AD converter control circuit 306. The AD converter 705 drives the control signal 708 in synchronization with the clock of the clock signal 707, and outputs optical signal data, which is a digital value resulting from the AD conversion, to the AD converter control circuit 306.

[0085] Each of the photodiodes 709 and 711 and the photomultiplier tube 710 outputs optical signal data to the AD converter control circuit 306 by a receiving circuit similar to that of the photomultiplier tube 701. In the configuration shown in Fig. 7 , the AD converter control circuit 306 controls the timing of the conversion start signal of each AD converter 705 to acquire the light intensity of the 670 nm fluorescent light and reflected light and the 780 nm fluorescent light and reflected light at any timing.

[0086] 8 is a timing chart showing the operational timing of main scanning, sub-scanning, height scanning, light irradiation, and optical signal data acquisition in the specimen measurement device of this embodiment. The horizontal axis of these timing charts represents time. Timing chart 801 shows the operational timing of main scanning, indicating that the piston crank mechanism 120 rotates at the times indicated by the hexagons, causing the main scanning of the light projector 104. X-axis position 802 shows the change in the position of the light projector 104 on the X-axis over time, with the vertical axis corresponding to the position of the light projector 104 in the X-axis direction and the horizontal axis representing time. The piston crank mechanism 120 causes the light projector 104 to reciprocate, and the speed is shown to be fast near the center of the stroke in the X-axis direction and slower near the stroke ends.

[0087] Timing chart 803 shows the operation timing of sub-scanning. The pulse motor 110 operates at the times indicated by hexagons, and sub-scanning is performed. Timing chart 804 shows the operation timing of height scanning. The pulse motor 113 operates at the times indicated by hexagons, and height scanning is performed. Timing chart 805 shows the operation timing of acquiring optical signal data when irradiated with 780 nm excitation light. The photomultiplier tube 701 and receiving circuit 106 operate at the times indicated by hexagons, and optical signal data derived from the 780 nm excitation light is acquired. Timing chart 806 shows the operation timing of acquiring optical signal data when irradiated with 670 nm excitation light. The photomultiplier tube 701 and receiving circuit 106 operate at the times indicated by hexagons, and optical signal data derived from the 670 nm excitation light is acquired. Timing chart 807 shows the operation timing of the 780 nm laser in the light source 102. At the times indicated by the hexagons, 780 nm excitation light is emitted and irradiated onto the array plate 101 via the light projecting unit 104. Timing chart 808 shows the operation timing of the 670 nm laser in the light source 102. At the times indicated by the hexagons, 670 nm excitation light is emitted and irradiated onto the array plate 101 via the light projecting unit 104.

[0088] In this manner, in this embodiment, when the X coordinate of the light-projecting unit 104 is moving in the forward direction, 780 nm excitation light is irradiated onto the array plate 101, and optical signal data generated by the 780 nm excitation light is acquired. On the other hand, when the X coordinate of the light-projecting unit 104 is moving in the backward direction, 670 nm excitation light is irradiated onto the array plate 101, and optical signal data generated by the 670 nm excitation light is acquired. In this manner, by acquiring optical signal data with different excitation wavelengths in the forward and backward directions, the influence of misalignment of images in the forward and backward directions, which occurs due to mechanical errors in the piston-crank mechanism 120, delay times in the receiving circuit 106, etc., is reduced.

[0089] 8, when the X coordinate of the main scanning acquired by the linear encoder 108 during forward movement reaches X1, acquisition of optical signal data based on the 780 nm excitation light is started. Then, when the X coordinate of the main scanning acquired by the linear encoder 108 during backward movement reaches X2, acquisition of optical signal data based on the 670 nm excitation light is started. When the X coordinate of the main scanning reaches X1 during backward movement, it is determined that acquisition of optical signal data in the backward direction is complete, a sub-scan trigger and a height scan trigger are output, and a sub-scan and height scan are performed to move to the next row.

[0090] When the X coordinate of the main scanning direction acquired by the linear encoder 108 during forward movement reaches X3, irradiation of the 780 nm excitation light is initiated. When the X coordinate of the main scanning direction reaches X4 during forward movement, it is determined that acquisition of the 780 nm optical signal data is complete, and irradiation of the 780 nm excitation light is stopped. Furthermore, when the X coordinate of the main scanning direction acquired by the linear encoder 108 during backward movement reaches X4, irradiation of the 670 nm excitation light is initiated. When the X coordinate of the main scanning direction reaches X3 during backward movement, it is determined that acquisition of the 670 nm optical signal data is complete, and irradiation of the 670 nm excitation light is stopped.

[0091] Next, the differences between the control method of the specimen measurement device 1 according to the present disclosure and the control method of specimen measurement devices according to the prior art will be explained using Figures 9A to 9F. Here, first, using Figures 9A and 9B, a problem that occurs when sampling of optical signal data is simply performed in synchronization with reaching a predetermined X coordinate in the main scanning direction, as in the prior art, will be explained.

[0092] FIG. 9A shows a timing chart of main scanning performed under ideal conditions with minimal vibration in the piston crank mechanism 120 and the like. Specifically, it shows the operational timing of the AD converter 705, AD converter control circuit 306, and data acquisition control circuit 312 of the receiving circuit 106. In FIG. 9A, the position of the light projector in main scanning is indicated by an X coordinate 901, but FIG. 9A is an enlarged view of the vicinity of the stroke center of the X-axis position 802 of the light projector 104 in FIG. 8. In FIG. 9A, the output signal from the linear encoder 108 is converted to an X coordinate by the coordinate calculation circuit 310 and shown as a timing chart 902 corresponding to the encoder coordinate. For convenience, the coordinate at the start point is set to 0 in FIG. 9A.

[0093] 9A shows the voltage of the conversion start signal 706 that the AD converter control circuit 306 outputs to the AD converter 705 of the receiving circuit 106. Here, the conversion start signal 706 rises every time the X coordinate of the light projecting unit 104 advances by 2 μm, as specified by Xp. When the AD converter 705 detects the rising edge of the voltage of the conversion start signal 706, it samples the fluorescence signal output from the filter circuit 704. Timing chart 904 shows the timing of signal sampling performed within the AD converter 705. The AD converter 705 operates and performs sampling at the times indicated by the hexagons in the timing chart 904. The sampled fluorescence signal is held in the AD converter 705 and converted to a digital value.

[0094] Timing chart 905 shows the timing of conversion of fluorescent light signals into digital values ​​that takes place within the AD converter 705. The AD converter 705 operates at times indicated by hexagons, indicating that conversion to digital values ​​is taking place. Timing chart 906 shows the timing at which the AD converter 705 outputs optical signal data to the AD converter control circuit 306. At times indicated by hexagons, a data signal 708 consisting of an optical signal is transmitted together with a clock signal 707, indicating that digital values ​​are being output. The AD converter control circuit 306 sequentially transfers the optical signal data acquired from the AD converter 705 to the memory 303. Timing chart 907 shows the timing at which the AD converter control circuit 306 transfers the digital values ​​received to the memory 303 via the bus 302. At times indicated by hexagons, the bus 302 operates, indicating that digital values ​​are being output.

[0095] The AD converter 705 is in operation from the start of sampling shown in timing chart 904 to the completion of output of digital values ​​shown in timing chart 906, and is therefore unable to start sampling the next fluorescent light signal. This period is referred to as the busy period of the AD converter. Timing chart 908 shows the busy period of the AD converter 705. During the busy period indicated by the arrow, the AD converter 705 is in operation, and this indicates that sampling cannot start even if the conversion start signal 706 is input during that period. In this embodiment, the length of this busy period is 800 ns.

[0096] 9A, there is little vibration during main scanning. In an ideal state, there is little sudden change in the slope of the X coordinate 901, indicating little variation in the main scanning speed. In this case, there is little variation in the timing of the conversion start signal, which is indicated by a change in voltage, and its rising edge is outside the busy period indicated by timing chart 908. Therefore, in this case, optical signal data is acquired without omission in synchronization with the X coordinate.

[0097] However, in reality, due to vibrations of the piston crank mechanism 120 and the like, the scanning of the light projector 104 does not result in a simple change in position. Figure 9B is a timing chart showing the operation timing of the AD converter 705, AD converter control circuit 306, and data acquisition control circuit 312 of the receiving circuit 106 when such main scanning vibrations are present. The dotted line X coordinate 901, like the X coordinate 901 in Figure 9A, indicates the change in the X coordinate of the main scanning in an ideal case. Note that timing charts 902 to 908 are the same as the timing charts 902 to 908 in Figure 9A, respectively, and therefore will not be described here. The solid line 909 indicates the actual change in the X coordinate of the main scanning when vibrations are present. Compared to the change shown by the dotted line as the X coordinate 901, it can be seen that the X coordinate vibrates in a complex manner at a high frequency in both positive and negative directions. In such a case, the main scanning is performed at the same average speed as in Figure 9A when viewed globally, but the speed varies significantly locally.

[0098] As a result, the coordinate values ​​of the linear encoder shown in timing chart 902 may fluctuate significantly, causing the irradiation position Pi on the X coordinate to advance 2 μm and pass the next target position Pt earlier than expected. In this case, as shown in FIG. 9B , the conversion start signal, represented by the rising voltage in timing chart 903, rises within the busy period shown in timing chart 908. According to the example of FIG. 9C , timings 910 and 911 in timing chart 904 are examples of when the conversion start signal is sent within the busy period. In such a case, the corresponding fluorescent light signal cannot be acquired, and the AD converter 705 cannot perform AD conversion, resulting in a failure to acquire optical signal data.

[0099] As described above, speed variations can occur in the main scanning due to vibrations, etc. If sampling is simply performed in synchronization with the X coordinate of the main scanning, as in conventional technology, the occurrence of such speed variations may lead to failure to acquire optical signal data due to the conversion start signal being output during a busy period of the AD converter.

[0100] The present disclosure has been made in view of the above circumstances, and an example of an optical signal data acquisition method according to a first embodiment will now be described with reference to FIGS. 9C to 9F.

[0101] 9C is a timing chart showing the operation timing of the AD converter 705, AD converter control circuit 306, and data acquisition control circuit 312 of the receiving circuit 106 in the ideal case where main scanning vibration is small in this embodiment. The present embodiment shown in FIG. 9C differs from the conventional example shown in FIG. 9A in the following respects. Specifically, the timing of the voltage rise of the conversion start signal 706 shown in timing chart 903 and the timing of data transfer from the AD converter control circuit 306 to the memory 303 shown by the hexagon in timing chart 907. Note that the change in X coordinate 901 and timing charts 902, 904, 905, 906, and 908 are the same as those described in FIG. 9A, and therefore will not be described here.

[0102] In the example of timing chart 903 shown in Fig. 9A, the timing of the voltage rise of the conversion start signal is synchronized with the coordinates output from the coordinate calculation circuit 310. In contrast, in this embodiment shown in Fig. 9C, the conversion start signal 706 operates so that the voltage rises at a constant frequency regardless of the coordinates output from the coordinate calculation circuit 310. Here, the frequency of the voltage rise is set to 1 MHz. This cycle is 1 us, which is longer than the length of the busy period, 800 ns, so the voltage rise of the conversion start signal 706 always occurs outside the busy period.

[0103] In the timing chart 907, in the example of FIG. 9A , the optical signal data after AD conversion is sequentially transferred to the memory 303 in response to acquisition of the optical signal data from the AD converter 705. In contrast, in the example of FIG. 9C , the operational timing of data transfer to the memory 303 is synchronized with the coordinates output from the coordinate calculation circuit 310. That is, when the X coordinate advances 2 μm and reaches the target position Pt, a data acquisition trigger signal is output from the synchronization circuit 311 to the data acquisition control circuit 312. The data acquisition control circuit 312 transfers the latest digital value that had been output to the ADC control circuit up to that point to the memory 303. In this way, although the AD conversion is performed asynchronously with the X coordinate of the main scanning shown in the timing chart 902, the transfer of the optical signal data is synchronized with the X coordinate, so that optical signal data closest to the initial target position Pt is transferred to the memory 303.

[0104] 9D is a timing chart showing the operation timing of the AD converter 705, AD converter control circuit 306, and data acquisition control circuit 312 of the receiving circuit 106 when main scanning vibration is present in this embodiment. The differences between the example of FIG. 9D and the example of FIG. 9B are the timing of the voltage rise of the conversion start signal 706 shown in timing chart 903, and the timing of data transfer from the AD converter control circuit 306 to the memory 303 shown in timing chart 907. Note that the change in X coordinate 901, timing charts 902, 904, 905, 906, 908, and solid line 909 are the same as those described in FIG. 9B, and therefore will not be described here.

[0105] In the example of timing chart 903 shown in FIG. 9B , the timing of the rising edge of the drive voltage of the conversion start signal is synchronized with the coordinates output from the coordinate calculation circuit 310. In contrast, in this embodiment shown in FIG. 9D , the conversion start signal 706 operates so that the voltage rises at a constant frequency regardless of the coordinates output from the coordinate calculation circuit 310. Here, the frequency of the voltage rise is set to 1 MHz. This cycle is 1 μs, which is longer than the length of the busy period, 800 ns. Therefore, even if the coordinates of the linear encoder reach the target position Pt during the busy period due to main scanning vibration, the voltage rise of the conversion start signal always occurs outside the busy period.

[0106] In the timing chart 907 shown in FIG. 9B , the AD-converted optical signal data is sequentially transferred to the memory 303 in response to the acquisition of the optical signal data from the AD converter 705. In contrast, in the example shown in FIG. 9D , the timing of data transfer to the memory 303 is synchronized with the coordinates output from the coordinate calculation circuit 310. That is, when the X coordinate of the irradiation position Pi of the primary light advances by 2 μm and the irradiation position Pi reaches the target position Pt, the synchronization circuit 311 outputs a data acquisition trigger signal to the data acquisition control circuit 312. The data acquisition control circuit 312 transfers the latest digital value that had been output to the ADC control circuit up to that point to the memory 303. As described above, in this embodiment, AD conversion is performed asynchronously with the X coordinate of the main scanning shown in the timing chart 902, but the transfer of the optical signal data is synchronized with the X coordinate. This configuration prevents the loss of optical signal data due to passing the target position Pt during a busy period, while transferring optical signal data that is close to the regular intervals of the original target position Pt to the memory 303.

[0107] An example where the scanning speed is fast near the center of the stroke at the X-axis position 802 in Fig. 8 has been described above using Fig. 9A to Fig. 9D. Below, an example where the scanning speed is slow near the end of the stroke at the X-axis position 802 in Fig. 8 will be described with reference to Fig. 9E and Fig. 9F.

[0108] 9E is a timing chart showing the operation timing of the AD converter 705, AD converter control circuit 306, and data acquisition control circuit 312 of the receiving circuit 106 when ideal main scanning is performed with little main scanning vibration in this embodiment. In Fig. 9E, the X coordinate 901 indicates the change in position in the main scanning, and is an enlarged view of the vicinity of the stroke end of the X-axis position 802 in Fig. 8. Compared to the example in Fig. 9C, the slope of the change in the X coordinate 901 is smaller because the scanning speed is slower.

[0109] The operation of timing charts 902 to 908 is the same as the example in FIG. 9C . That is, the voltage rise of the conversion start signal 706 operates at a constant frequency regardless of differences in the moving speed of the X coordinate. Here, the frequency of the voltage rise is assumed to be 1 MHz. This cycle is 1 μs, which is longer than the length of the busy period, 800 ns, so the voltage rise of the conversion start signal always occurs outside the busy period.

[0110] In the example of FIG. 9E , in the timing chart 907, the operation timing of data transfer to the memory 303 is synchronized with the coordinates output from the coordinate calculation circuit 310. That is, when the X coordinate of the irradiation position Pi of the primary light advances by 2 μm and the irradiation position Pi reaches the target position Pt, the synchronization circuit 311 outputs a data acquisition trigger signal to the data acquisition control circuit 312. The data acquisition control circuit 312 transfers the latest digital value output to the ADC control circuit up to that time to the memory 303. Note that, unlike the example of FIG. 9C , when the scanning speed is slow, AD conversion may be performed on the optical signal data for the next scanning position before the previous optical signal data is transferred to the memory 303. In this case, the data acquisition control circuit 312 overwrites the old optical signal data with new optical signal data and retains only the latest optical signal data. The optical signal data obtained at the timing indicated by the hatched hexagon in the timing chart 906 is overwritten and therefore not transferred to the memory. As a result, the number of optical signal data transferred to the memory 303 is smaller than the number of optical signal data to be AD converted.

[0111] In this way, by transferring only the latest optical signal data to the memory 303 in synchronization with the X coordinate, only optical signal data in which the irradiation position Pi of the primary light is closest to the target position Pt is selected and stored in the memory 303. Therefore, the capacity of the memory 303 and the amount of data communication are reduced compared to when AD conversion is simply performed at a constant frequency and the acquired optical signal data is sequentially stored in the memory 303. Furthermore, when AD conversion is simply performed at a constant frequency and the acquired optical signal data is sequentially stored in the memory 303, the acquisition interval of the optical signal data varies greatly depending on the scanning speed. However, the method of this embodiment acquires optical signal data at nearly equal intervals even when the scanning speed changes significantly.

[0112] FIG. 9F is a timing chart showing the operation timing of the AD converter 705, AD converter control circuit 306, and data acquisition control circuit 312 of the receiving circuit 106 when main scanning vibrations are present in this embodiment. The dotted line shown as the X coordinate 901 represents the X coordinate of the main scanning in an ideal case, similar to the X coordinate 901 in FIG. 9E. Timing charts 902 to 908 are the same as the timing charts 902 to 908 in FIG. 9E, respectively, and therefore will not be described here. The solid line 909 represents the actual change in the X coordinate of the main scanning when vibrations are present. Compared to the change in the X coordinate 901 shown by the dotted line, it can be seen that the X coordinate vibrates in a complex manner at a high frequency in both positive and negative directions. In such a case, the main scanning is performed at the same average speed as in FIG. 9E when viewed globally, but the speed variation is greater when viewed locally.

[0113] As a result, in the timing chart 902, the timing at which the irradiation position Pi of the primary light passes through the target position Pt is shifted, and the relationship between the timing of the voltage rise of the data acquisition trigger signal and the timing of the conversion start signal is different from the ideal case shown in FIG. 9E. As a result, the optical signal data transferred to the memory 303 differs between the example case of FIG. 9E and the example case of FIG. 9F. More specifically, when there is no vibration, the optical signal data third from the left in the timing chart 906 of FIG. 9E is transferred to the memory 303. In contrast, when there is vibration, the optical signal data second from the left in the timing chart 906 of FIG. 9F is transferred to the memory 303. In this way, even when there is vibration, optical signal data close to the target position Pt can be selectively transferred to the memory, and optical signal data at nearly equal intervals can be acquired.

[0114] Such local vibrations are difficult to predict in advance because they change not only due to position and speed but also due to deterioration over time, etc. The optical signal data acquisition method according to this embodiment reflects such changing conditions, prevents omission of optical signal data, and acquires optical signal data at nearly equal intervals, which is the initial target position Pt.

[0115] Here, the process of acquiring height information of the array plate 101 in the specimen measurement device 1 in this embodiment will be described with reference to Figure 10 and Figures 11A to 11C. Figure 10 shows the operational flow for acquiring height information of the array plate 101 using the specimen measurement device 1 according to this embodiment. What differs from the photographing operation is that when acquiring height information, sub-scanning is not performed, but height scanning is performed at a constant pitch while acquiring reflected light signal data, which is then analyzed to calculate the height of the array plate surface. The photodiode 709 in the optical sensor 105 is used to acquire the height information.

[0116] 11A to 11C are diagrams illustrating the positional relationship of height scanning of the array plate during height information acquisition operation. Figures 11A and 11B are side views of the array plate 101. Figure 11C plots the amount of reflected light acquired by the optical sensor 105 during height scanning for each height, with the horizontal axis representing the amount of light and the vertical axis representing the acquired height.

[0117] When the height information acquisition process starts, first in step S1001, the CPU 301 sets parameters for acquiring height information in the synchronization circuit 311. The set parameters include the Y coordinate Yh of the position where height information acquisition is performed, the pixel pitch Xp in the main scanning direction, and the pixel pitch Zp in the height scanning direction. The set parameters also include points 1101 (X5, Z5) and 1102 (X6, Z6) that indicate the height scanning range on the XZ plane, and the rotation speed Xs in the main scanning direction.

[0118] In this embodiment, a rectangular region 1103 with points 1101 and 1102 on the XZ plane as diagonal corners is referred to as the height scanning region. Here, the number of pixels in the main scanning direction Nx = (X6 - X5) / Xp and the number of pixels in the height scanning direction Nz = (Z6 - Z5) / Zp are calculated in advance. In this embodiment, X5 = 500, X6 = 22,500, Z5 = 2,000, Z6 = 6,000, Xp = 10 μm, Zp = 10 μm, and Xs = 1,200 rpm. In this case, Nx = 2,200 and Nz = 400. After the parameters are set, the flow proceeds to step S1002.

[0119] In the height information acquisition process, unlike the imaging process, the intervals between acquisitions of optical signal data may be large. Therefore, the time required for height information acquisition and the data volume are reduced. In step S1002, the CPU 301 issues instructions to the motor control circuits 307, 308, and 309 to move the array plate 101 and the light projector 104 to the height information acquisition start position. In this embodiment, the X coordinate of the height information acquisition start position is the end of the scanning range of the piston crank mechanism 120, and the X coordinate value is -2500. The Y coordinate of the height information acquisition start position is Yh specified by the parameter. The Z coordinate of the height information acquisition start position is Z5 specified by the parameter. After the movement is completed, the flow proceeds to step S1003.

[0120] In step S1003, the CPU 301 starts main scanning in the same manner as in step S405 of the image capturing process, and then the flow proceeds to step S1004. In step S1004, the CPU 301 starts irradiating the light source 102 with 760 nm excitation light, and then the flow proceeds to step S1005. Note that in step S1004, irradiation with 670 nm excitation light is not performed.

[0121] In step S1005, the synchronization circuit 311 determines whether the light-projecting unit 104 has reached the line feed position. Specifically, the synchronization circuit 311 determines that the line feed position has been reached when the X coordinate of the light-projecting unit 104 output from the coordinate calculation circuit 310 moves from inside the image capture area to outside the image capture area. If the current main scanning direction is the forward direction (the direction in which the X coordinate increases), the synchronization circuit 311 determines that the line feed position has been reached when the X coordinate of the light-projecting unit 104 exceeds X6. Scanning in the forward direction is represented by the trajectory 1104 of the light-projecting unit 104 shown in FIG. 11B. Furthermore, if the current main scanning direction is the backward direction (the direction in which the X coordinate decreases), the synchronization circuit 311 determines that the line feed position has been reached when the X coordinate of the light-projecting unit 104 becomes smaller than X5. Scanning in the backward direction is represented by the trajectory 1106 of the light-projecting unit 104 shown in FIG. 11B. The initial value of the main scanning direction is the forward direction, and thereafter, each time the stroke end is reached, the scanning direction is repeatedly changed back and forth. This type of scanning is sometimes referred to as boustrophedonic scanning.

[0122] Unlike the photographing process, the height information acquisition process requires only one wavelength of light irradiation. Therefore, line feeds can be performed on both the forward and backward passes, shortening the time required to acquire height information. When the light projecting unit 104 reaches the line feed position, the synchronization circuit 311 outputs a height scan trigger signal, and the flow proceeds to step S1014. When the light projecting unit 104 has not reached the line feed position, the synchronization circuit 311 does not output a height scan trigger signal, and the flow proceeds to step S1006.

[0123] In step S1006, the determination unit 173 incorporated in the synchronization circuit 311 determines whether the irradiation position Pi has reached the target position Pt, similar to step S409 of the imaging process. The target positions Pt are multiple points on a locus, such as point 1108 illustrated in FIG. 11B, arranged at a pitch of Xp in the X direction and a pitch of Zp in the Z direction. If the synchronization circuit 311 determines that the light projecting unit 104 has reached the target position Pt, the synchronization circuit 311 outputs a data acquisition trigger signal, and the flow proceeds to step S1007. If the synchronization circuit 311 determines that the light projecting unit 104 has not yet reached the target position Pt, the synchronization circuit 311 does not output a data acquisition trigger signal, and the flow proceeds to step S1009.

[0124] In step S1007, the AD converter control circuit 306 and the data acquisition control circuit 312 execute the same process as in step S410 of the image capture process to acquire optical signal data, and the flow proceeds to step S1008. However, here, the data acquisition control circuit 312 sets its internal data acquisition completion register to 1 after storing Nx×Nz pieces of optical signal data in the memory 303. The data acquisition control circuit 312 sets its internal data acquisition completion register to 0 when Nx×Nz pieces of optical signal data have not been stored in the memory 303. After acquiring the optical signal data, the flow proceeds to step S1008. In step S1008, the synchronization circuit 311 updates the target position Pt stored internally, similar to step S411 of the image capture process, and the flow proceeds to step S1009.

[0125] In step S1009, the CPU 301 determines whether or not the acquisition of optical signal data has ended, similarly to step S412 of the photographing process. If the CPU 301 determines that the acquisition of optical signal data has ended, the flow proceeds to step S1010. If the CPU 301 determines that the acquisition of optical signal data has not ended, the flow proceeds to step S1005.

[0126] In step S1010, the CPU 301 issues an instruction to the light source control circuit 305 to stop the light emission of the light source 102, similar to step S413 of the photographing process, and the flow proceeds to step S1011. In step S1011, the CPU 301 issues an instruction to the motor control circuit 307 to stop the rotation of the pulse motor 107 and end the main scanning, similar to step S414 of the photographing process, and the flow proceeds to step S1012. In step S1012, the CPU 301 issues an instruction to the motor control circuits 307, 308, and 309 to move the array plate 101 and the light projector 104 to their stop positions, similar to step S415 of the photographing process, and the flow proceeds to step S1013.

[0127] In step S1013, the CPU 301 reads out the Nx×Nz pieces of optical signal data stored in the memory 303, analyzes them, and calculates height information for the array plate. Specifically, the Nx pieces of optical signal data acquired at the same height are averaged to determine the average light intensity for each height. When the calculated average light intensity is arranged by Z coordinate, two peaks corresponding to the front and back surfaces of the array plate 101 are present, as shown in FIG. 11C . More specifically, in FIG. 11C , peak 1109 indicates a peak due to light reflected from the front surface of the array plate 101, and peak 1110 indicates a peak due to light reflected from the back surface of the array plate 101. Of these peaks, peak 1109 with the larger Z coordinate, i.e., Z coordinate 1111 indicating the peak corresponding to the front surface, is set as height information corresponding to position Yh. Once the height information is obtained, the height information acquisition process is terminated.

[0128] In step S1014, the CPU 301 issues an instruction to the motor control circuit 309 to move the array plate 101 by Zp in the height scanning direction. Here, if the amount of movement of the array plate 101 when one pulse of a voltage pulse signal is sent to the motor driver 114 is Mz, the number of pulses output by the motor control circuit 309 to the motor driver 114 is Zp / Mz. The height scanning is represented by trajectories 1105 and 1107 in FIG. 11B, and the movement distance in the Z direction is Zp. Based on the instruction from the CPU 301, the motor control circuit 309 outputs a pulse signal to the motor driver 114 at a speed such that the height scanning is completed while the light projecting unit 104 is outside the imaging area in the main scanning direction. When the height scanning corresponding to one pulse is completed, the flow proceeds to step S1015.

[0129] In step S1015, the synchronization circuit 311 increments the current Z coordinate by Zp from the previous Z coordinate, and the flow proceeds to step S1009. When the tilt information acquisition process in the shooting process is executed in step S402, the height information acquisition process shown in Fig. 10 is executed twice for the Y coordinates Y3 and Y4 to determine the Z coordinates Z3 and Z4 for each, and the tilt information K is calculated using equation 1.

[0130] In this embodiment, the imaging region 503 covers the entire region 204, and an example has been described in which all spots on the array plate 101 are imaged, but the present invention is not limited to this. That is, the user can set any part of the region 204 as the imaging region 503. This allows scanning only a part including the spot of interest, thereby reducing the imaging time.

[0131] In this embodiment, height information is obtained using the peak position of reflected light from the surface of the glass slide, but the method of obtaining height information is not limited to this. Height information may also be obtained using the peak position of the brightness of fluorescent signals from some spots on the array plate. In this case, it is necessary to irradiate some spots with light to obtain height information, but the number of components in the optical system can be reduced because there is no need to obtain reflected light with an optical sensor.

[0132] In this embodiment, an example has been described in which two wavelengths from the light source are switched between the forward and backward paths, but the method of controlling the light source is not limited to this. It is also possible to simultaneously irradiate excitation light of two wavelengths on both the forward and backward paths, and separate and acquire the fluorescence derived from each excitation light using an optical system. In this case, two rows are advanced in one round trip, thereby reducing the imaging time by approximately half.

[0133] The analyte measurement device according to the present disclosure, described above as the first embodiment, optically measures analyte characteristics using an array substrate on which spots are arranged in an array. The analyte measurement device 1 includes a light irradiating means, a light detecting means, a scanning means, a position acquiring means, a converting means, a first holding means, a second holding means, a determining means, a memory means, and a transferring means. The present disclosure also provides an analyte measurement method in which these means are operated as follows. In this embodiment, the light projecting unit 104 constitutes one aspect of the light irradiating means of the present disclosure, which irradiates the array substrate (array plate 101) with concentrated primary light (excitation light with wavelengths of 670 nm and 780 nm) for a predetermined period of time. The optical sensor 105 constitutes one aspect of the light detecting means of the present disclosure, which detects secondary light, which is fluorescence from the spots 202 irradiated with the primary light, at a predetermined period of time and outputs optical signal data as a detection signal. The piston crank mechanism 120, the linear stage 109, and the motors and the like associated therewith constitute an aspect of the scanning means of the present disclosure, which changes the irradiation position of the primary light relative to the array substrate. The coordinate calculation circuit 310 constitutes an aspect of the position acquisition means 138 of the present disclosure, which acquires information about the irradiation position of the primary light on the array substrate. The receiving circuit 106 including the AD converter 705 and the AD converter control circuit 306 constitute an aspect of the conversion means 146 of the present disclosure, which converts the detection signal acquired during a predetermined period into digital data. Note that the predetermined period described here corresponds to the period defined by the encoder coordinates 0 and 1 shown in the timing chart 902 illustrated in FIG. 9F.

[0134] In the present disclosure, the first holding means 171 is included as a buffer memory in the synchronization circuit 311 as shown in FIG. 17 . That is, the synchronization circuit 311 calculates a target position Pt, which is set as a position for acquiring optical signal data with respect to the irradiation position of the primary light, and stores the calculated target position Pt in a buffer memory disposed in the synchronization circuit 311, constituting one aspect of the first holding means 171 in the present disclosure. Note that the X coordinate P(N) of the Nth target position Pt is expressed as X1 + Xp × N (Xp: pitch in the X direction). However, the target position Pt does not necessarily have to be stored by the synchronization circuit 311, but may also be stored by a coordinate calculation circuit 310 that calculates the X coordinate of the light projector 104 based on the output of the linear encoder 108. The second holding means 172 is included as a buffer memory in the synchronization circuit 311 as shown in FIG. 17 and stores digital data converted by the conversion means 146 shown in FIG. 18 . The memory 303 constitutes a storage means in the present disclosure, in which the digital data held in the second holding means 172 and transferred by a transfer means described later is stored in association with information about the irradiation position Pi. Therefore, the memory 303 may be referred to as a storage means that stores the irradiation position Pi and the digital data in association with each other.

[0135] Furthermore, the determination means 173 determines whether the irradiation position Pi of the primary light has reached the target position Pt based on information about the relative position obtained from the linear encoder 108 or the like, constituting one aspect of the determination means in the present disclosure. The data acquisition control circuit 312 selects digital data corresponding to the data output during a predetermined period in FIG. 9F (the two hexagons on the left side of the timing chart 906) in accordance with the determination result by the determination means 173 that the irradiation position Pi has reached the target position Pt. The memory control circuit 304 transfers the digital data selected in this manner to the memory 303, constituting one aspect of the transfer means in the present disclosure. Note that although the data acquisition control circuit 312 and the memory control circuit 304 are disclosed as separate components in the present disclosure, they can also be treated as an integrated component. In other words, the data acquisition control circuit 312 is a transfer means that selects the digital data stored in the second storage means 172 and transfers it to the storage means (memory 303) in accordance with the determination result by the determination means 173. The selected digital data is the digital data held in the second holding means 172 and corresponds to the irradiation position Pi corresponding to the target position Pt.

[0136] The analyte measurement device 1 may further include a light source 102 and a light source control circuit 305. The light source 102 is optically coupled to the light irradiating means and emits primary light that is guided to the light projecting unit 104. The light source control circuit 305 constitutes the light source control means in this embodiment and controls the on / off of the light emitted by the light source 102. The light irradiating means in this embodiment also includes an objective optical system 129 that focuses the primary light on the array plate 101. The light detecting means includes an optical sensor 105 that constitutes the light detecting unit in this embodiment and is optically coupled to the objective optical system 129 to detect secondary light guided from the spot on the array plate 101. As described above, the scanning means also includes a piston crank mechanism 120, a linear stage 109, and associated motors, etc. In this embodiment, the piston crank mechanism 120 includes a rotation drive unit 121 and a piston crank unit 123. The rotary drive unit 121 is configured with a motor or the like and rotates a drive shaft, constituting one aspect of the rotary drive unit of the present disclosure. The rotary drive unit is connected to one end of a crank 118, which is one end of a piston crank unit 123, and converts the rotational motion of the rotary shaft into one-dimensional reciprocating motion. The other end of a connecting rod 119, one end of which is connected to the other end of the crank 118, is connected to the other end of the piston crank unit 123, to which an objective optical system 129 (light projector 104) is connected. With this structure, the scanning means is able to reciprocate the objective optical system (see FIG. 16 ).

[0137] In this embodiment, as shown by the spot positions in FIG. 2A, the Pt are arranged at equal intervals on the array substrate (array plate 101). Information regarding the target positions Pt is stored as data in the first storage unit 171. The data Itp related to the target positions Pt stored in the first storage unit 171 is updated in response to the movement or relative change of the irradiation position by the main scanning unit. The conversion unit (AD converter 705) converts the detected optical signal data into digital data within a predetermined period shorter than the time required for the main scanning unit to scan the primary light between adjacent target positions Pt. The transfer unit (data acquisition control circuit 312) preferentially selects and transfers to the memory 303 the digital data closest to the end of the predetermined period. In this embodiment, the data of the unhatched hexagon second from the left in the data output timing chart 906 in FIG. 9F (immediately before number 2 in the encoder coordinate timing chart 902) is selected and transferred to the memory 303. Although the immediately preceding data is most preferable as the data to be transferred, it is also possible to use a buffer memory or the like to select the immediately preceding two data, the data immediately before the immediately preceding data, and transfer these.

[0138] The specimen measurement device 1 according to the present disclosure further includes an imaging unit (image processing unit 301a) that images specimen characteristics based on digital data stored in a storage unit (memory 303). The specimen measurement device 1 uses the digital data assigned to each pixel (target position Pt) by the image processing unit 301a to image the specimen characteristics and displays the image on the user interface 117. Furthermore, the specimen measurement device 1 according to the present disclosure further includes a buffer memory that temporarily stores the converted digital data. In this embodiment, such a buffer memory is provided in the data acquisition control circuit 312. When multiple converted digital data are obtained within the time required for the primary light irradiation position to scan consecutive target positions Pt, the temporarily stored digital data is overwritten by the digital data obtained immediately thereafter. In the example shown in FIG. 9F, in the timing chart 906 showing data output, the data indicated by the leftmost hatched hexagon is overwritten by the data indicated by the next white hexagon. This process is performed by the data acquisition control circuit 312.

[0139] The present disclosure can also be understood as a control method for a sample measurement device. This control method includes a light irradiation step, a light detection step, a scanning step, a position acquisition step, and a conversion step, all of which are executed by the controller circuit 116 using the above-described configuration, and further includes a determination step and a transfer step. The light irradiation step is a process executed continuously from step S406 in FIG. 4 onward, in which focused primary light is irradiated onto the array substrate for a predetermined period. The light detection step is a process executed in step S410, in which secondary light from the spot is detected at a predetermined cycle and a detection signal is output from the optical sensor 105. The scanning step is a process executed continuously from step S405 onward, in which the irradiation position of the primary light on the array substrate is changed. The position acquisition step is a process executed in conjunction with the scanning step in step S405, in which information about the irradiation position is acquired. The conversion step is a process executed in step S410, in which the detection signal acquired by the optical sensor 105 at a predetermined cycle is converted into digital data. The determination step is a process executed in step S409, and determines whether the irradiation position Pi corresponds to the target position Pt. The transfer step selects data corresponding to the timing at which the irradiation position Pi reaches the target position Pt from the digital data held in the second holding means 172, depending on the determination result of the determination step, and transfers the data to the storage means (memory 303). By repeatedly executing each of these steps, it becomes possible to obtain digital data in a manner that enables the generation of an image of the sample.

[0140] As described above, in the specimen measurement device 1 according to this embodiment, AD conversion of the signal from the optical sensor is performed at regular intervals. This prevents omission of optical signal data even when the irradiation position Pi passes the target position Pt during AD conversion due to local speed variations in the main scanning, ensuring stable acquisition of fluorescent signals. Meanwhile, the transfer of acquired optical signal data to memory is synchronized with the position of the light projector. This allows optical signal data acquired at positions close to the target position Pt at equal intervals to be stored in memory, even when using a drive mechanism with large position-dependent speed variations, such as the piston-crank mechanism 120. Furthermore, there is no need for a high-speed AD converter to accommodate local speed variations in the main scanning, or a robust scanning mechanism to reduce speed variations. This eliminates the need for a high-speed AD converter to accommodate local speed variations in the main scanning, and achieves high-precision measurement and reduced imaging time while preventing increased costs for the specimen measurement device.

[0141] Second Embodiment A second embodiment of the present disclosure will be described below. The second embodiment differs from the first embodiment in that, when acquiring optical signal data, optical signal data is acquired in a number greater than the number of pixels, and a single pixel data set is generated from the acquired multiple optical signal data sets. More specifically, in the first embodiment, the pixel positions and the target position Pt corresponded one-to-one, but there was a possibility that the influence of random noise from the photomultiplier tube would cause variations in image brightness. Therefore, in this embodiment, a large number of optical signal data sets are acquired at intervals finer than the pixel pitch, and the data sets are averaged to generate a single pixel data set, thereby reducing the influence of random noise from the photomultiplier tube and improving measurement accuracy and image quality.

[0142] The configuration of the specimen measurement device, the configuration of the array plate, and the internal functional configuration of the controller circuit in this embodiment are the same as those illustrated in Figures 1, 2A, 2B, and 3, respectively. Therefore, the same reference numerals will be used and a description thereof will be omitted. Below, the operational flow for photographing an array plate using the specimen measurement device of this embodiment will be described using Figures 5A, 5B, 6A, 6B, 12, and 13.

[0143] 12 shows the operational flow when photographing the array plate 101 using the specimen measurement device 1 according to this embodiment. When the user inputs an instruction to start photographing the array plate 101 via the user interface 117, processing from step S1201 onwards begins. In step S1201, the CPU 301 reads the photographing conditions specified by the user via the user interface 117 and stores them in the memory 303. The CPU 301 also sets information based on the photographing conditions in various circuits within the controller circuit 116.

[0144] As imaging conditions, a point 501 (X1, Y1) and a point 502 (X2, Y2) indicating the imaging range on the array plate 101, a pixel pitch Xp in the main scanning direction, a pixel pitch Yp in the sub-scanning direction, and a rotation speed Xs in the main scanning direction are input. In this embodiment, in addition to these, an oversampling rate Rx is also input. Note that the oversampling rate in this embodiment corresponds to the number of optical signal data (digital data obtained from detection signals) acquired to generate one pixel. If Rx=5, one pixel data is generated from five optical signal data acquired at intervals of 1 / 5 of the pixel pitch Xp.

[0145] In this embodiment, it is assumed that X1 = 500, X2 = 22,500, Y1 = 500, Y2 = 64,500, Xp = 10 μm, Yp = 10 μm, Xs = 1,200 rpm, and Rx = 5 are input. In this case, the sampling pitch in the main scanning direction is Sx = 2 μm. Here, a rectangular area 503 with points 501 and 502 on the array plate 101 as diagonals is referred to as the imaging area. The number of pixels in the main scanning direction, Nx = (X2 - X1) / Xp, and the number of pixels in the sub-scanning direction, Ny = (Y2 - Y1) / Yp, are calculated. In this embodiment, according to the above settings, Nx = 2,200 and Ny = 6,400. Once the above settings are complete, the flow proceeds to step S1202.

[0146] Here, the processes executed in steps S1202 to S1208 are the same as the processes executed in steps S402 to S408 described in the first embodiment, and therefore, a description of the processes executed in these steps will be omitted here.

[0147] In step S1209, the synchronization circuit 311 determines whether the light-projecting unit 104 has reached the target position Pt. Point 507 in FIG. 5B indicates the position of a pixel to be imaged on the array plate 101. In this embodiment, the target position is a point on the array plate 101 at which optical signal data is acquired in order to generate a pixel, and the X coordinate P(N) of the Nth target position Pt is expressed by the following equation 6: P(N)=X1+(Xp / Rx)×N (N=0, 1, ..., Nx×Rx-1) (Equation 6). The target positions are multiple points on a trajectory, with a pitch in the X direction of Xp / Rx and a pitch in the Y direction of Yp.

[0148] The positional relationship between the target position and the pixel position will now be described with reference to FIG. 13 . FIG. 13 is an enlarged view of a portion of FIG. 5B . Points 1301, 1302, and 1303, indicated by circles, are pixel positions and are the same as point 507 in FIG. 5B . The distance between points 1301 and 1302 is Xp, and the distance between points 1301 and 1303 is Yp. Points 1304 to 1308, indicated by crosses, are target positions, and are evenly spaced apart by Xp / Rx, centered on pixel w. In this embodiment, this distance is 2 μm. In this embodiment, optical signal data is acquired at five locations from target positions 1304 to 1308, and the luminance value of pixel 1302 is generated by averaging. In other words, the number of optical signal data acquired initially is Rx times the number of pixels.

[0149] The initial value of the target position Pt is P(0), which is stored in a buffer memory or the like provided inside the synchronization circuit 311. In determining the first target position Pt, it is determined that the target position Pt has been reached when the X coordinate of the light-projecting unit 104 output from the coordinate calculation circuit 310 passes through P(0) in the forward direction (the direction in which the X coordinate increases). In determining the target position Pt from the second time onwards, it is determined that the target position Pt has been reached when the X coordinate of the light-projecting unit 104 output from the coordinate calculation circuit 310 passes through the target position Pt for which the irradiation position Pi was updated in step S1211, which will be described later.

[0150] If the synchronization circuit 311 determines that the target position Pt has been reached, the synchronization circuit 311 outputs a data acquisition trigger signal, and the flow proceeds to step S1210. If the synchronization circuit 311 determines that the target position Pt has not yet been reached, the synchronization circuit 311 does not output a data acquisition trigger signal, and the flow proceeds to step S1212.

[0151] In step S1210, the AD converter control circuit 306 outputs a conversion start signal, and the output voltage from the optical sensor 105 is amplified by the receiving circuit 106, filtered, and then AD converted. In this embodiment, the conversion start signal is a signal with a constant period, which is longer than the conversion time of the AD converter but shorter than the time it takes for the light projecting unit 104 to move the distance Xp / Rx. The AD-converted optical signal data is temporarily stored in a buffer memory inside the data acquisition circuit 312. The buffer memory can store one optical signal data per AD converter, and the latest optical signal data is overwritten and saved each time AD conversion is performed. In this embodiment, an AD converter corresponding to the wavelength of each light source is provided, and optical signal data reflecting the amount of fluorescent light excited by the wavelength of each light source is saved.

[0152] When the data acquisition control circuit 312 receives a data acquisition trigger signal, it reads out the optical signal data stored in the buffer memory and outputs it to the memory control circuit 304 via an internal DMA controller and the bus 302. The memory control circuit 304 stores the optical signal data at a specified address in the memory 303. The address in the memory 303 is incremented each time a data acquisition trigger signal is received.

[0153] After storing Nx×Rx×Ny pieces of optical signal data corresponding to the wavelengths of each light source in memory 303, the data acquisition control circuit 312 sets an internal data acquisition completion register to 1. When the Nx×Rx×Ny pieces of data corresponding to the wavelengths of each light source have not been stored in memory 303, the data acquisition control circuit 312 sets an internal data acquisition completion register to 0. Once the optical signal data at target position Pt has been stored, the flow proceeds to step S1211.

[0154] Note that the processes executed in steps S1211 to S1215 in this embodiment are similar to the processes executed in steps S411 to S415 in the first embodiment, and therefore, a description of the processes executed in these steps will be omitted here.

[0155] In step S1216, the CPU 301 reads Rx×Nx×Ny pieces of optical signal data corresponding to the wavelengths of each light source from the memory 303 and performs image formation processing. In the image formation processing, the Rx pieces of optical signal data in the main scanning direction are averaged to generate one pixel data. The five pieces of optical signal data from points 1304 to 1308 are averaged to generate pixel data for point 1302, which is then stored in the memory 303. As a result of the image formation processing, the number of pixel data corresponding to each light source wavelength becomes Nx×Ny. The optical signal data is separated into signal components reflecting the amount of fluorescent light and random noise components derived from the optical sensor and receiving circuit. By performing averaging, the noise components can be reduced to 1 / √Rx while the signal components remain the same, leading to an improved S / N ratio. After the CPU 301 performs image formation processing for each light source wavelength, the flow proceeds to step S1217.

[0156] Next, in step S1217, the CPU 301 reads out Nx x Ny pieces of pixel data corresponding to the wavelengths of each light source from the memory 303, performs data compression and format conversion, and creates a captured image file in TIFF format. The captured image file is saved in the memory 303 and presented to the user via the UI control circuit 314 and the user interface 117. In addition, the captured image file can be transferred to an external data server via the communication circuit 313 in response to an instruction from the user.

[0157] The processes executed in steps S1218 to S1222 in this embodiment are the same as the processes executed in steps S417 to S421 in the first embodiment, and therefore, a description of the processes executed in these steps will be omitted here.

[0158] In this embodiment, oversampling is performed in the main scanning direction to acquire optical signal data at intervals finer than the pixel pitch, but oversampling is not performed in the sub-scanning direction to acquire optical signal data at intervals equal to the pixel pitch. This prevents an increase in imaging time. In this embodiment, the oversampling rate Rx is input by the user via the user interface 117. However, the acquisition method is not limited to this; for example, the rate can be obtained from the memory 303 based on predetermined conditions. The memory 303 functions as an imaging condition storage unit in this embodiment, which stores the oversampling rate Rx, which corresponds to the number of digital data pieces to be acquired to generate one pixel, input as an imaging condition. In this embodiment, the user interface 117 functions as an input unit that receives a command for the oversampling rate Rx from an intermediary unit. In this embodiment, the image formation process and the averaging process performed during this process have been described using an example in which the image processing unit 301a in the CPU 301 functions as an averaging unit. Specifically, in this embodiment, the image processing unit 301a serves as an averaging unit, averaging multiple pieces of digital data stored in the memory 303 to generate pixel data indicative of the characteristics of the specimen. However, the entity that performs image formation or image processing is not limited to this example. An image formation circuit may be implemented downstream of the data acquisition control circuit 312 in the FPGA of the controller circuit 116, and image formation processing may be performed by hardware. In this case, the number of pieces of data can be reduced by averaging before transferring the optical signal data to the memory 303, resulting in effects such as reduced memory capacity and reduced imaging time.

[0159] Although the present embodiment has been described using an example in which simple averaging is performed in the image formation process, a more complex image formation algorithm may also be used. Weighted addition processing may be used to increase the weight of the optical signal data of point 1306, which is close to the position of point 1302, and decrease the weight of the optical signal data of points 1304 and 1308, which are far from the position of point 1302.

[0160] As described above, in the imaging process according to this embodiment, AD conversion is performed at regular intervals, but the transfer of acquired optical signal data to memory is performed in synchronization with the position of the light projector. This allows a large amount of optical signal data to be acquired stably at short intervals, even when vibration occurs in the main scanning direction. Furthermore, by averaging these data to generate a single pixel data, the effects of noise from the optical sensor and receiving circuit are reduced, improving measurement accuracy and image quality.

[0161] Third Embodiment A third embodiment of the present disclosure will be described below. The third embodiment differs from the second embodiment in that the voltage range of the optical signal data is adjusted before averaging in the image formation process, and the pixel data value is adjusted after averaging.

[0162] The configuration of the analyte measurement device, the configuration of the array plate, and the internal functional configuration of the controller circuit in this embodiment are the same as those illustrated in Figures 1, 2A, 2B, and 3, respectively. Therefore, the same reference numerals are used, and their descriptions are omitted here. The configurations of the optical sensor 105 and the controller circuit 116 are the same as those described in the first embodiment, and the operational flow for performing imaging is also the same as that described in the second embodiment. Hereinafter, the same reference numerals are used for the same configurations or processes, and this embodiment will be described using Figures 7, 12, and 14A to 14C.

[0163] FIG. 14A shows the relationship between the output voltage of the current-voltage conversion amplifier 702 (see FIG. 7 ) and the digital value of the output of the AD converter 705 in the third embodiment. In the figure, the horizontal axis represents time, and the vertical axis represents voltage. In FIG. 14A , output voltage 1401 is the output voltage of the current-voltage conversion amplifier 702 when the amount of light incident on the optical sensor 105 is large. When the amount of incident light is large, the output voltage 1401 becomes a positive voltage. On the other hand, output voltage 1402 is the output voltage of the current-voltage conversion amplifier 702 when the amount of light incident on the optical sensor 105 is weak. On average, the output voltage 1402 is close to 0 V, but may become a slightly negative voltage due to voltage offsets in the optical sensor and receiving circuit, etc. In this case, if only positive voltages are assigned from 0 to 65535 and averaged, the negative voltages will be clamped to 0, and the averaged voltage will be slightly offset in the positive direction.

[0164] Therefore, in the third embodiment, the level conversion circuit 703 inside the receiving circuit 106 converts the range 1403 indicated by the arrow, which can cover the output voltages 1401 and 1402, from 0 V to 5 V. Here, the upper limit of the range 1403 is Tmax, and the lower limit is Tmin. Tmax and Tmin are determined in advance. In this embodiment, Tmax = 10 V, and Tmin = -0.3 V. Since the AD conversion range is from 0 V to 5 V, the relationship between the input voltage Vin and the output voltage Vout of the level conversion circuit 703 is expressed as shown in Equation 7 below: Vout = 5 × (Vin - Tmin) / (Tmax - Tmin) (Equation 7). In this embodiment, the resistance values ​​of the resistor network of the level conversion circuit 703 are set to input and output values ​​that satisfy the condition shown in Equation 7. Furthermore, the digital value D output by the AD converter 705 is expressed as shown in Equation 8 below. D=(Vout / 5)×65535=(Vin-Tmin) / (Tmax-Tmin)×65535 (Formula 8)

[0165] 14B is a diagram showing the relationship between input and output from the voltage level conversion inside the receiving circuit 106 to the AD converter. In this diagram, the horizontal axis represents the input voltage Vin of the level conversion circuit, and the vertical axis represents the digital value after AD conversion. In the receiving circuit 106 of this embodiment, level conversion and AD conversion are performed so that −0.3 V becomes the digital value 0, and 10 V becomes the digital value 65535. By providing such level conversion, even signals whose output voltage is less than 0 V due to the offset voltage of the optical sensor 105 and the receiving circuit 106 when the amount of light is weak can be acquired without leakage.

[0166] However, if the averaged pixel data is imaged as is, even if the average value is 0 V, the digital value will be larger by the amount of the 0.3 V offset. Therefore, even in areas where there is no signal, the brightness will be increased accordingly, and areas in the captured image where there are no spots 202 on the array plate 101 will also be bright. The imaging process of this embodiment differs from the imaging process of the second embodiment in that, in the image formation process of step S1216 of FIG. 12 , after averaging to calculate the pixel data at the target position Pt, level conversion is again performed on each pixel data to adjust the output pixel data. The processes other than step S1216 of FIG. 12 are the same as the processes executed in each step of the second embodiment, and therefore will not be described here.

[0167] Here, in the image formation process of step S1216, the pixel data after level conversion is assumed to be L. If the pixel data when the average value of the voltage Vin is 0 V is Pzero, then Pzero = -Tmin / (Tmax - Tmin) x 65535. In this embodiment, Tmax = 10 V and Tmin = -0.3 V, so Pzero = 1908. FIG. 14C is a diagram showing the input / output relationship of pixel level conversion in the image formation process. In this diagram, the horizontal axis represents pixel data at the target position Pt after averaging, and the vertical axis represents output pixel data L after pixel level conversion. In the image formation process of this embodiment, linear conversion is performed so that the output pixel data L = 0 when Pt ≦ Pzero, and L = 65535 when Pt = 65535. In other words, when Pt is equal to or greater than Pzero, the relationship between L and Pt satisfies the relationship shown in Equation 9 below. L=65535 / (65535-Pzero)*(Pt-Pzero) (Equation 9) If Pt is less than Pzero, L becomes 0. Then, the output pixel data is stored in memory 303, and the flow proceeds to step S1217.

[0168] In this embodiment, an example has been described in which the maximum value remains the same before and after pixel-level conversion. However, if the maximum value of the pixel data after averaging becomes smaller than 65,535 due to saturation of the receiving circuit or the like, the maximum value may also be converted in the image formation process. Furthermore, if the maximum value of the pixel data after averaging is limited to 62,000 due to saturation within the receiving circuit or the like, the range of 1,908 to 62,000 of the pixel data of the target position Pt after averaging may be assigned to the output pixel data L from 0 to 65,535.

[0169] That is, in this embodiment, the image processing unit 301a, as averaging means, averages digital data converted from detection signals in a first range including positive and negative detection signals, as exemplified by range 1403 in Fig. 14A, using the AD converter control circuit 306. Then, the image processing unit 301a performs level conversion on the averaged data, assigning a part of the first range to pixel values.

[0170] As with the averaging process executed in the second embodiment, the pixel level conversion process in this embodiment can also be performed by hardware by implementing an image forming circuit after the data acquisition control circuit 312 in the FPGA of the controller circuit 116. In this case, there are advantages such as a reduction in the amount of data transferred to the memory 303 and a reduction in the shooting time.

[0171] As described above, in the imaging process according to this embodiment, when oversampling is performed, the range of A / D conversion of the optical signal is widened to also acquire negative values ​​near 0 V. This prevents the average value of the optical signal data from being offset in the positive direction when the amount of light incident on the optical sensor 105 is weak. Furthermore, by performing pixel-level conversion again after averaging, areas of the captured image where there is no signal are prevented from becoming bright.

[0172] Fourth Embodiment A fourth embodiment of the present disclosure will be described below. The fourth embodiment differs from the first embodiment in that the target position Pt is shifted between the forward and backward paths so as to reduce the deviation between a predetermined target position and the position at which the optical signal data actually stored in memory is sampled.

[0173] The configuration of the array plate and the internal functional configuration of the controller circuit of the specimen measurement device in this embodiment are the same as those illustrated in Figures 1, 2A, 2B, and 3, respectively. Therefore, the same reference numerals are used, and their description will be omitted. The configurations of the optical sensor 105 and the controller circuit 116 are the same as those described in the first embodiment, and the operational flow for capturing images is also the same as that described in the first embodiment. Hereinafter, the same reference numerals will be used for the same configurations or processes, and this embodiment will be described using Figures 1, 7, 9F, 15, and 16.

[0174] 9F, the trigger period of the AD conversion in timing chart 903 is denoted by Ta, and the delay time from when linear encoder 108 detects that light projector 104 has actually passed a certain X coordinate until data transfer is performed is denoted by Td1. Furthermore, the delay time from when AD converter 705 performs sampling as shown in timing chart 904 until it outputs optical signal data as shown in timing chart 906 is denoted by Td2. The moving speed of light projector 104 is denoted by Vx. In this case, Vx corresponds to the slope of the dotted line shown as X coordinate 901.

[0175] The optical signal data in timing chart 907 that is transferred to memory is optical signal data that has been sampled and AD converted before the irradiation position Pi passes through the target position Pt. The second optical signal data from the left in Fig. 9F is transferred when the coordinate of the light-projecting unit 104 is 2, but in reality it was sampled when the encoder coordinate is 1.

[0176] The difference dt between the time when the irradiation position Pi passes the target position Pt and the time when the optical signal data stored in memory is sampled is between (Td2 - Td1) and (Td2 - Td1 + Ta), taking into account the delay time. The deviation between the predetermined target position and the position when the optical signal data stored in memory is actually sampled is ideally dt multiplied by Vx, and is a value between (Td2 - Td1) x Vx and (Td2 - Td1 + Ta) x Vx.

[0177] Note that Td1 is a constant value determined by the linear encoder 108, the coordinate calculation circuit 310, the synchronization circuit 311, and the data acquisition control circuit 312, and is set to 250 ns in this embodiment. Td2 is a constant value determined by the receiving circuit 106 and the AD converter control circuit 306, and is set to 800 ns in this embodiment. Furthermore, if the frequency of the AD converter 705 is 1 MHz, Ta is 1 μs. Vx is a value that varies between 0 m / s and 2 m / s depending on the rotation angle of the piston crank mechanism 120. Limited to the imaging region 503, Vx varies between 1 m / s and 2 m / s. In this case, the deviation of the target position Pt is 0.55 μm to 1.55 μm when Vx is near 1 m / s on the forward path, but increases to approximately 1.1 μm to 3.1 μm when Vx is near 2 m / s. The deviation has a range because the sampling and the position of the light-projecting unit 104 are asynchronous, and varies depending on the timing within the cycle Ta at which the irradiation position Pi passes the target position Pt. On the return path, the amount of position deviation is the same but the direction of deviation is reversed. Therefore, in this embodiment, the target position Pt is shifted to correct the fixed portion of the position deviation, (Td2-Td1) x Vx.

[0178] FIG. 15 is a diagram showing a target position Pt in this embodiment. Points 1301, 1302, and 1303 in FIG. 15 are pixel positions, and are the same as the pixel positions in the second embodiment described with reference to FIG. 13. Points 1304 to 1308 indicated by an "x" are points evenly spaced at intervals of Xp / Rm around a pixel, and are the same as the target positions described with reference to FIG. 13. Point 1511 indicated by an "x" is a point obtained by shifting point 1304 by Xp / Rm in the backward direction, and point 1512 is a point obtained by shifting point 1308 by Xp / Rm in the forward direction.

[0179] In Figure 15, the points indicated by squares are target positions when the light projector 104 is moving in the forward direction, and are target positions for acquiring fluorescent signal data for 760 nm excitation light. When generating pixels for the captured image at point 1302 based on the 760 nm excitation light, an arithmetic average of optical signal data sampled at five positions from point 1501 to point 1505 is used. The points indicated by triangles are target positions when the light projector 104 is moving in the backward direction, and are target positions for acquiring fluorescent signal data for 670 nm excitation light. When generating pixels for the captured image at point 1302 based on the 670 nm excitation light, an arithmetic average of optical signal data sampled at five positions from point 1506 to point 1510 is used.

[0180] Note that the points indicated by squares, crosses, and triangles are shifted in the Y-axis direction for clarity, but in reality, the Y coordinates of all three are the same. The target position indicated by squares is shifted in the backward direction from the position indicated by crosses by (Td2-Td1) x Vx. Similarly, the target position indicated by triangles is shifted in the forward direction from the position indicated by crosses by (Td2-Td1) x Vx. That is, in this embodiment, the target position is set to be shifted between the forward and backward directions depending on the delay time that occurs during conversion to digital data by the AD converter control circuit 306. This corrects position deviations resulting from delay times that occur in the receiving circuit 106 and the controller circuit 116 by generating a data acquisition trigger earlier than the actual position to be acquired.

[0181] In this embodiment, the amount of positional deviation is calculated as follows: Referring to Fig. 16, which is an enlarged view of the periphery of piston-crank mechanism 120 shown in Fig. 1, the position x of light projecting unit 104 is expressed by the following equation 10 using the length r of crank 118, the length l of connecting rod 119, and the angle θ of pulse motor 107.

[0182] Moreover, the velocity Vx is expressed by the following equation 11 by differentiating equation 10.

[0183] θ is calculated by multiplying the rotation angle per pulse by the number of drive pulses. Furthermore, since r and l are known values, the speed of the light projector 104 is calculated for each drive pulse using Equation 11, and by multiplying this by the delay time Td2-Td1, the positional deviation amount for each X coordinate on the forward and backward paths is determined in advance. The determined positional deviation amount is stored in the synchronization circuit 311. However, because the rotation angle per pulse is in increments of 0.72°, the obtained x value is also discrete. Therefore, in this embodiment, the coordinate calculation circuit 310 uses the angular velocity to interpolate and estimate the coordinates between pulses.

[0184] As described above, in the imaging process according to this embodiment, the target position Pt is shifted between the forward and backward paths in advance, taking into consideration the speed of the piston crank mechanism 120 and the delay times of the linear encoder 108, the receiving circuit 106, and the controller circuit 116. This reduces the positional deviation between the predetermined target position and the position at which the optical signal data actually stored in memory is sampled, making it possible to acquire optical signal data with high accuracy.

[0185] Although the present embodiment has been described using an example in which optical signal data is acquired by shifting the position during the sampling stage, other methods may be used to correct positional deviations during the forward and backward passes. The target position is the same for the forward and backward passes, as indicated by the cross marks in FIG. 15 , but the optical signal data used to generate pixels during the forward pass and the backward pass in the image formation process may be shifted. When generating pixel data for point 1304, the five optical signal data sets from point 1511 and points 1304 to 1307 may be averaged during the forward pass, and the five optical signal data sets from point 1305 to point 1308 and point 1512 may be averaged during the backward pass. With this method, the effect of correcting positional deviations is limited by the optical signal data acquisition interval Xp / Rm, but it provides a simple method for correcting positional deviations during the forward and backward passes.

[0186] Other Embodiments The present disclosure can also be realized by providing software (programs) that realize one or more functions of the various embodiments described above to a system or device via a network or a storage medium, and having a computer in the system or device read and execute the programs. The computer has one or more processors or circuits, and may include multiple separate computers or a network of multiple separate processors or circuits to read and execute executable instructions on the computer.

[0187] In this case, the processor or circuit may include a central processing unit (CPU), a microprocessing unit (MPU), or a graphics processing unit (GPU). The processor or circuit may also include an application specific integrated circuit (ASIC), or a field programmable gate array (FPGA). The processor or circuit may also include a digital signal processor (DSP), a data flow processor (DFP), or a neural processing unit (NPU).

[0188] As described above, the specimen measurement device according to the present disclosure is low-cost and can stably acquire optical signals even when variations in scanning speed occur. As a result, a large number of optical signals are acquired at short intervals. As a result, it is possible to achieve high measurement accuracy and a reduction in imaging time while preventing an increase in the cost of the device.

[0189] Although the present disclosure has been described above with reference to embodiments and modifications, the present disclosure is not limited to the above embodiments and modifications. Inventions modified within the scope of the present disclosure and inventions equivalent to the present disclosure are also included in the present disclosure. Furthermore, the above-described embodiments and modifications may be combined as appropriate within the scope of the present disclosure.

[0190] The present invention is not limited to the above-described embodiments, and various modifications and variations can be made without departing from the spirit and scope of the present invention. Therefore, the following claims are appended to apprise the public of the scope of the present invention.

[0191] This application claims priority based on Japanese Patent Application No. 2023-212900 filed on December 18, 2023 and Japanese Patent Application No. 2024-186737 filed on October 23, 2024, the entire contents of which are incorporated herein by reference.

[0192] 1: specimen measuring device 129: objective optical system (122, 125) 122: light irradiation means (104, 103, 102) 125: light detection means (104, 103, 105) 130: scanning means (130x, 130y, 130z) 130x: main scanning means (107, 120, 118, 119) 130y: sub-scanning means (112, 113, 114) 138: position acquisition means (108, 310) 146: conversion means (751, 306) 171: first holding means (311, modified example 303) 172: second holding means (311, modified example 303) 173: determination means (311) 304: transfer means

Claims

1. A specimen measurement device which optically measures characteristics of a specimen using an array substrate on which spots are provided in an array, comprising: light irradiating means for irradiating concentrated primary light onto said array substrate for a predetermined period of time; light detecting means for detecting secondary light from said spots at a predetermined cycle and outputting a detection signal; scanning means for changing the irradiation position of said primary light on said array substrate; position acquiring means for acquiring information regarding said irradiation position; conversion means for converting the detection signal detected at said predetermined cycle into digital data; first holding means for holding information of a target position Pt related to said irradiation position; second holding means for holding said digital data; 2. The specimen measurement device according to claim 1, further comprising an imaging condition storage means for storing, as an imaging condition, an oversampling rate corresponding to the number of digital data acquired to generate one pixel.

3. The specimen measurement device according to claim 2, further comprising an input section for receiving a command for the oversampling rate from outside.

4. A specimen measurement device as described in any one of claims 1 to 3, comprising a light source optically coupled to the light irradiation means, and a light source control means for controlling the on / off of the light emission of the light source.

5. A specimen measurement device as described in any one of claims 1 to 4, wherein the light irradiating means comprises an objective optical system that focuses the primary light on the array substrate, and the light detecting means comprises a light detecting section that is optically coupled to the objective optical system and through which the secondary light is guided.

6. The specimen measurement device according to claim 5, wherein the scanning means comprises a rotary drive unit and a piston crank unit which converts the rotary motion of the rotary drive unit into one-dimensional reciprocating motion and causes the objective optical system to reciprocate.

7. The specimen measurement device according to any one of claims 1 to 6, wherein the target positions Pt are arranged at equal intervals on the array substrate.

8. A specimen measurement device as described in any one of claims 1 to 7, wherein the target position Pt held by the first holding means is updated based on the movement or change of the irradiation position.

9. A specimen measurement device according to any one of claims 1 to 8, wherein the conversion means converts the detection signal into digital data in a period shorter than the time required for the primary light to scan between adjacent target positions Pt.

10. A specimen measurement device as claimed in any one of claims 1 to 9, wherein the transfer means preferentially selects from the digital data that is closest to the end of the specified period and transfers it to the storage means.

11. The specimen measurement device according to any one of claims 1 to 10, further comprising imaging means for imaging characteristics of the specimen based on the digital data stored in the memory means.

12. A specimen measurement device according to any one of claims 1 to 11, further comprising an averaging means for averaging a plurality of digital data stored in the memory means to generate pixel data indicative of a characteristic of the specimen.

13. The specimen measurement device according to claim 12, wherein the averaging means performs level conversion on the averaged digital data converted by the conversion means from a detection signal in a first range including positive and negative detection signals, thereby assigning a portion of the first range to pixel values.

14. A specimen measurement device as described in any one of claims 1 to 13, wherein the scanning means scans the irradiation position of the primary light back and forth on the array substrate, and the target position Pt is set to be shifted between the forward and return paths according to a delay time that occurs during conversion to digital data by the conversion means.

15. A specimen measurement device as described in any one of claims 1 to 14, further comprising a buffer memory for temporarily storing the converted digital data, wherein when a plurality of converted digital data are obtained in the buffer memory within the time required for the irradiation position of the primary light to scan consecutive target positions Pt, the temporarily stored digital data is overwritten by the digital data obtained immediately thereafter.

16. A control method for a specimen measurement device including first holding means for holding a target position related to the irradiation position of primary light, second holding means for holding detection signals detected by measurement of secondary light from the spots as digital data, and memory means for storing the irradiation position in association with the digital data, in order to optically measure characteristics of a specimen using an array substrate on which spots are provided in an array, the control method for a specimen measurement device including a light irradiation step of irradiating the array substrate with concentrated primary light for a predetermined period of time, a light detection step of detecting secondary light from the spots at predetermined cycles and outputting detection signals, a scanning step of changing the irradiation position of the primary light on the array substrate, a position acquisition step of acquiring information regarding the irradiation position, and a conversion step of converting the detection signals acquired at the predetermined cycles into digital data, a determination step of determining whether the irradiation position corresponds to the target position Pt, and a transfer step of selecting data corresponding to the timing when the irradiation position reached the target position Pt from the digital data held in the second holding means in accordance with the determination result determined in the determination step, and transferring the data to the memory means.

17. A program for causing a computer to execute each step of the method for controlling a specimen measurement device according to claim 16.

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