Scanning device, scanning method, and program

Abstract

To acquire focused optical information in a short period.SOLUTION: A scanning device includes: an observation optical system that radiates primary light toward one surface in order to acquire optical information related to at least some of a plurality of spots; a scanning section that performs main scanning in which the observation optical system moves relative to an array plate 101 in a first direction and acquires the optical information, and sub-scanning in which the observation optical system moves relative to the array plate 101 in a second direction intersecting the first direction without acquiring the optical information; and an adjustment section that adjusts the relative position of the observation optical system with respect to the array plate 101 in an optical axis direction of the primary light. The adjustment section performs the adjustment if the scanning section is in a sub-scanning period.SELECTED DRAWING: Figure 4

Description

Technical Field

[0001] The present invention relates to a scanning device, a scanning method, and a program.

Background Art

[0002] There are known protein array plates or peptide array plates on which a large number of biological substances having peptide bonds such as proteins and peptides are immobilized on a substrate. Using this, it is possible to perform interactions with a large number of biological substances immobilized on the substrate at once. Such array plates are effective for comprehensively analyzing the interactions between a liquid specimen derived from a living body, such as blood, cell extract, saliva, interstitial fluid, etc., and a large number of proteins or peptides. By such analysis, the characteristics of the specimen can be measured.

[0003] The immobilization site of a protein or peptide on the substrate is called a spot. As a method for observing a spot that has received an interaction with a specimen, for example, there is known a method of identifying which spot has received an interaction by labeling the spot with a fluorescent probe. A microarray scanner is known as a device for observing an array plate labeled with a fluorescent probe.

[0004] Patent Document 1 discloses a microarray scanner having an irradiation optical system, a fluorescence detection optical system, and a two-dimensional scanning system. The irradiation optical system has a function of condensing and irradiating a laser beam onto an array plate. The fluorescence detection optical system has a function of detecting the amount of fluorescence from a spot labeled with a fluorescent probe. The two-dimensional scanning system has a function of obtaining a fluorescence image of spots on the array plate by two-dimensionally scanning the array plate or the optical system. Also, a confocal optical system is used as the fluorescence detection optical system. Patent Document 2 discloses a scanning optical device that simplifies the position adjustment in the height direction when obtaining a fluorescence image.

Prior Art Documents

Patent Documents

[0005] [Patent Document 1] U.S. Patent No. 7911670 [Patent Document 2] Patent No. 5281756 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] Because array plates have individual differences in glass thickness and tilt, there is a problem in confocal optical systems with a shallow depth of focus where it is difficult to obtain a fluorescence image that is in focus across the entire array plate.

[0007] Patent Document 1 describes an automatic focus adjustment using a focus sensor simultaneously with two-dimensional scanning to obtain a fluorescence image that is in focus across the entire array plate. However, accurately performing automatic focus adjustment while rapidly scanning the array plate or optical system in two dimensions requires high-speed feedback control consisting of a high-performance focus sensor and a low-vibration actuator, which makes the device complex.

[0008] In Patent Document 2, obtaining a fluorescence image that is in focus across the entire array plate requires repeating two-dimensional scanning multiple times while changing the collection position and setting parameters, which results in a long fluorescence measurement time.

[0009] This invention has been made in view of the above-mentioned problems, and aims to acquire focused optical information in a short amount of time. [Means for solving the problem]

[0010] The present invention relates to a scanning apparatus for scanning an observation optical system with respect to an array plate having a plurality of spots on one surface, comprising: an observation optical system that irradiates primary light toward the one surface to acquire optical information relating to at least a portion of the plurality of spots; a scanning unit that performs a main scan in which the observation optical system moves relative to the array plate in a first direction and acquires the optical information; and a sub-scan in which the observation optical system moves relative to the array plate in a second direction intersecting the first direction without acquiring the optical information; and an adjustment unit that adjusts the relative position of the observation optical system with respect to the array plate in the optical axis direction of the primary light, wherein the adjustment unit Based on the information relating to the imaging area from which the optical information is acquired and the information relating to the position of the observation optical system, if the position of the observation optical system is outside the imaging area, The adjustment is performed when the scanning unit is in the sub-scanning period. [Effects of the Invention]

[0011] According to the present invention, in-focus optical information can be acquired in a short time. [Brief explanation of the drawing]

[0012] [Figure 1] This is a schematic diagram showing the configuration of the sample measuring device according to the first embodiment. [Figure 2] This is a diagram showing the configuration of an array plate. [Figure 3] This diagram shows the internal configuration of the controller according to the first embodiment. [Figure 4] This is a flowchart showing the operation of the shooting process in the first embodiment. [Figure 5] This figure shows the positional relationship of the sub-scan during the imaging process in the first embodiment. [Figure 6] This figure shows the positional relationship of height scanning in the imaging process of the first embodiment. [Figure 7] This is a flowchart showing the operation for obtaining height information in the first embodiment. [Figure 8] This diagram shows the positional relationship of the height scan during the height information acquisition operation. [Figure 9]It is a diagram showing the positional relationship of sub-scanning in the imaging process of the second embodiment. [Figure 10] It is a flowchart showing the operation of the imaging process of the second embodiment. [Figure 11] It is a diagram showing the internal configuration of the controller of the third embodiment. [Figure 12] It is a diagram for explaining the operation of the piston crank mechanism. [Figure 13] It is a flowchart showing the operation of the coordinate calculation circuit of the third embodiment.

Mode for Carrying Out the Invention

[0013] Hereinafter, preferred embodiments of the present invention will be described with reference to the accompanying drawings. <First Embodiment> FIG. 1 is a schematic diagram showing the configuration of the specimen measuring device 100 of the first embodiment. The specimen measuring device 100 functions as a scanning device for scanning an object. The specimen measuring device 100 measures a specimen as an object located on one surface of the array plate 101. The array plate 101 has a large number of biological substances immobilized at each spot on the slide glass and is fluorescently labeled.

[0014] The light source 102 is a semiconductor laser that emits light with a wavelength of around 670 nm. 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 of the array plate 101 and the reflected light from the surface (upper surface) of the array plate 101 to the optical sensor 105. The confocal optical system 103 is composed of a pinhole, a filter, a dichroic mirror, a quarter-wave plate, a deflection beam splitter, and a lens. By using the confocal optical system 103, the influence of the fluorescence component by the slide glass itself of the array plate 101 can be reduced, and the signal-to-noise ratio in the measurement of the fluorescence component derived from the spots can be increased. [[ID=​The light-emitting unit 104 irradiates a spot on the array plate 101 with excitation light. The light-emitting unit 104 consists of 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. In this embodiment, the light-emitting unit 104 is positioned below the array plate 101 and irradiates the excitation light upward. The light-emitting unit 104 is configured to irradiate the object with excitation light as primary light and collect fluorescence as secondary light. The light-emitting unit 104 corresponds to an example of an observation optical system. The optical sensor 105 converts light into an electrical signal. The optical sensor 105 can use a photomultiplier tube, a photodiode, or the like. The optical sensor 105 can separate and acquire fluorescence from a spot on the array plate 101 and reflected light from the surface of the array plate 101. The optical sensor 105 corresponds to an example of a detection unit that acquires optical information.

[0016] The piston-crank mechanism 106 moves the light-emitting unit 104 along a plane perpendicular to the optical axis of the lens of the light-emitting unit 104 or the optical axis of the excitation light. Specifically, the piston-crank mechanism 106 causes the light-emitting unit 104 to reciprocate in the short-side direction of the array plate 101. As the light-emitting unit 104 reciprocates, the excitation light from the light source 102 is scanned in the short-side direction of the array plate 101. The short-side direction of the array plate 101 is called the main scanning direction, and the reciprocating scanning by the piston-crank mechanism 106 is called the main scan. That is, the main scanning direction is parallel to the short side of the array plate 101. In this embodiment, the stroke of the main scan is approximately 30 mm. The operating direction of the light-emitting unit 104 itself is limited to the main scanning direction by a guide (not shown). The pulse motor 107 rotates the piston crank mechanism 106 at high speed. In this embodiment, the rotational speed of the pulse motor 107 is approximately 1200 rpm. The pulse motor 107 corresponds to an example of a drive unit.

[0017] The encoder 108 measures the position of the light-emitting unit 104 in the main scanning direction. The encoder 108 is installed on the piston-crank mechanism 106 and outputs phase difference pulse voltages consisting of A-phase, B-phase, and Z-phase according to the position of the light-emitting unit 104 in the main scanning direction. The encoder 108 corresponds to an example of a measurement unit.

[0018] The linear stage 109 moves the array plate 101 along a plane perpendicular to the optical axis of the lens of the light-emitting unit 104 or the optical axis of the excitation light. Specifically, the linear stage 109 moves the array plate 101 in a direction perpendicular to the main scan in the horizontal plane. The linear stage 109 is composed of a ball screw, a home sensor, etc. Scanning in a direction perpendicular to the main scan in the horizontal plane is called sub-scanning. That is, the sub-scanning direction is parallel to the long side of the array plate 101. The linear stage 109 has a mounting section for placing the array plate 101. The user places the array plate 101 on the mounting section in advance. The 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. The piston crank mechanism 106, pulse motor 107, linear stage 109, and pulse motor 110 correspond to an example of a scanning unit.

[0019] The motor driver 111 is a driver circuit for rotating the pulse motor 110. In this embodiment, when one pulse signal is input to the motor driver 111, the pulse motor 110 rotates by 0.72°, and the array plate 101 moves 2um in the sub-scanning direction.

[0020] The linear stage 112 moves the array plate 101 in a vertical direction along the optical axis of the lens of the light-emitting unit 104 or the optical axis of the excitation light. The linear stage 112 consists of a ball screw, a home sensor, etc. Vertical scanning is called height scanning. The pulse motor 113 is connected to the linear stage 112. The rotational motion of the pulse motor 113 is converted into linear motion by the ball screw of the linear stage 112. The linear stage 112 and pulse motor 113 correspond to an example of an adjustment unit.

[0021] The motor driver 114 is a driver circuit for rotating the pulse motor 113. In this embodiment, when one pulse signal is input to the motor driver 114, the pulse motor 113 rotates 0.72°, and the array plate 101 moves 1um upward in the vertical direction.

[0022] The motor driver 115 is a driver circuit for rotating the pulse motor 107. In this embodiment, when one pulse signal is input to the motor driver 115, the pulse motor 107 rotates by 0.72°, and the light-emitting unit 104 moves along the main scanning direction.

[0023] The controller 116 controls the entire sample measurement device 100. The controller 116 is composed of an FPGA, CPU, memory, etc., and functions as a computer. The controller 116 controls the light source 102, motor driver 111, motor driver 114, and motor driver 115 to perform main scanning, sub-scanning, and height scanning of the excitation light on the array plate 101. Simultaneously with scanning, the controller 116 acquires fluorescence signal data (two-dimensional image) based on the position information of the light-emitting unit 104 measured by the encoder 108 and the output signal from the light sensor 105, and stores it in its internal memory.

[0024] As will be described later, the controller 116 acquires height and tilt information of the array plate 101 and generates drive pulse trains to be output to the motor drivers 111 and 114 during sub-scanning and height scanning. The controller 116 also controls the timing of sub-scanning and height scanning, and the timing of acquiring fluorescence signal data, in synchronization with the position information of the light-emitting unit 104. Through this control, the thickness and tilt of the array plate 101 are corrected when the light-emitting unit 104 is outside the imaging area on the array plate 101, and height scanning can be performed so that the entire array plate 101 is in focus.

[0025] The user interface 117 is an interface for receiving instructions from the user and displaying the results. The user interface 117 consists of a keyboard, mouse, display, etc. The controller 116 can receive shooting instructions from the user via the user interface 117 and present image data based on fluorescence signal data to the user. Furthermore, the user can specify the shooting area, main scanning direction, and sub-scanning direction pixel pitch via the GUI on the user interface 117 during shooting.

[0026] The piston-crank mechanism 106 includes a crank 118 and a connecting rod 119. The crank 118 is connected to the rotation shaft and connecting rod 119 of the pulse motor 107 via joints. The length of the crank 118 is denoted as r. The connecting rod 119 is connected to the crank 118 and the light-emitting unit 104 via joints, respectively. The length of the connecting rod 119 is l.

[0027] Figure 2(a) is a view of the array plate 101 from above, and Figure 2(b) is a view of the array plate 101 from the side. The array plate 101 consists of a rectangular glass slide 201 having a short side and a long side, and numerous spots 202 arranged on its upper surface. A biomaterial containing peptide bonds is immobilized on each spot 202. In this case, one type of biomaterial is immobilized on each spot 202.

[0028] In this embodiment, the diameter of spot 202 is approximately 100 μm, and the spot spacing is 200 μm. The length of the array plate 101 in the short direction (short side) is 25 mm, and the length of the long direction (long side) is 75 mm. The upper left point 203 of the array plate 101 is the origin, the rightward direction in the short direction is the positive direction of the X axis, and the downward direction in the long direction is the positive direction of the Y axis. If the units of the X and Y coordinates are μm, the coordinates of the four corners of the array plate 101 are (0,0), (25000,0), (0,75000), and (25000,75000).

[0029] In this embodiment, the stroke of the piston crank mechanism 106 is 30 mm, which is 5 mm longer than the length of the array plate 101. That is, during the main scan, a range 2.5 mm longer to the left and right of the array plate 101 is scanned, and the X coordinate of the scanning range is between -2500 and 27500. Due to the convenience of spot creation and the user's gripping, the microscope slide 201 has areas where spot 202 exists and areas where it does not. Area 204 is the area on the microscope slide 201 where spot 202 exists. In this embodiment, the coordinates of the four corners of area 204 are (2000,2000), (23000,2000), (2000,65000), and (23000,65000). The user can specify the range of the Y coordinate for the sub-scan.

[0030] Figure 3 is a block diagram showing the internal configuration of the controller 116 in the first embodiment. The CPU 301 executes the software (program) that controls the entire controller 116. The CPU 301 consists of a microprocessor, cache memory, etc. The CPU 301 corresponds to an example of a control unit. The bus interface 302 is an interface for connecting the CPU 301 to various peripheral circuits.

[0031] Memory 303 stores imaging conditions input by the user, parameters of the sample measurement device 100, and fluorescence signal data. Memory 303 can use DDR4-SDRAM, SSD, etc. Memory corresponds to an example of a storage unit. The memory control circuit 304 controls the memory 303 based on access commands to the memory 303 via the bus interface 302.

[0032] The light source control circuit 305 is a control circuit for the CPU 301 to control the light source 102. The light source control circuit 305 consists of an interface conversion circuit, a DA converter, and the like. The CPU 301 can control the on / off state and light intensity of the laser irradiation of the light source 102 via the light source control circuit 305. The data acquisition circuit 306 is a circuit that acquires fluorescence signal data based on the output signal from the light sensor 105 and the relative position of the light-emitting unit 104 with respect to the array plate 101, based on instructions from the CPU 301, and continuously stores it in the memory 303. The data acquisition circuit 306 consists of a buffer circuit, an AD converter, an AD converter control circuit, a DMA controller, etc. The data acquisition circuit 306 corresponds to an example of an image acquisition unit.

[0033] The motor control circuit 307 generates a control signal for the motor driver 115 for the pulse motor 107, which is the main scanning motor, based on instructions from the CPU 301. The motor control circuit 307 generates a drive pulse voltage according to the instructions from the CPU 301 for the rotational speed, acceleration, displacement, rotational direction, and rotation start timing of the pulse motor 107. The motor control circuit 308 generates a control signal for the motor driver 111 for the pulse motor 110, which is a sub-scanning motor, based on instructions from the CPU 301. The motor control circuit 308 generates a drive pulse voltage according to the instructions from the CPU 301 for the rotational speed, acceleration, displacement, rotational direction, and rotation start timing of the pulse motor 110. The motor control circuit 309 generates a control signal to the motor driver 114 for the pulse motor 113, which is a height scanning motor, based on instructions from the CPU 301. The motor control circuit 309 generates a drive pulse voltage according to the instructions from the CPU 301 for the rotational speed, acceleration, displacement, rotational direction, and rotation start timing of the pulse motor 113.

[0034] The coordinate calculation circuit 310 counts two phase difference pulse signals, A-phase and B-phase, from the encoder 108 and calculates the position of the light-emitting unit 104. In this embodiment, the resolution of the encoder 108 is set to 1 μm. The coordinate calculation circuit 310 increases the coordinate of the light-emitting unit 104 by 1 μm when the level of either the A-phase signal or the B-phase signal changes and the A-phase signal is ahead of the B-phase signal. Conversely, the coordinate calculation circuit 310 decreases the coordinate of the light-emitting unit 104 by 1 μm when the level of either the A-phase signal or the B-phase signal changes and the B-phase signal is ahead of the A-phase signal.

[0035] The synchronization circuit 311 generates trigger signals for the data acquisition circuit 306, motor control circuit 308, and motor control circuit 309 based on the coordinate information calculated by the coordinate calculation circuit 310. Here, the trigger signal to the data acquisition circuit 306 is called the data acquisition trigger signal, the trigger signal to the motor control circuit 308 is called the sub-scan trigger signal, and the trigger signal to the motor control circuit 309 is called the height scan trigger signal. When the data acquisition circuit 306 receives a data acquisition trigger signal, it controls its internal AD converter to acquire one output data from the optical sensor 105 and stores it in the memory 303 via its internal DMA controller. When the motor control circuit 308 receives a sub-scan trigger signal, it outputs a drive pulse train to the motor driver 111 that corresponds to the amount of movement equal to the pixel pitch in the sub-scan direction. The pixel pitch in the sub-scan direction is specified by the user at the start of shooting and is stored in the memory 303. When the motor control circuit 309 receives the height scanning trigger signal, it outputs a drive pulse train corresponding to the amount of movement in the height scanning direction to the motor driver 114. The amount of movement in the height scanning direction is calculated by the CPU 301. The calculation method will be described later.

[0036] The communication circuit 312 is a circuit for connecting the sample measurement device 100 to an external network. The communication method uses a communication protocol that conforms to the Ethernet standard. By connecting the sample measurement device 100 to an external PC or server, it is possible to remotely control the imaging process and save data to external high-capacity storage.

[0037] The UI circuit 313 is a circuit for connecting the sample measuring device 100 to the user interface 117. The UI circuit 313 consists of input circuits for the keyboard and mouse, and an image forming circuit for controlling the display. The peripheral circuits constituting the controller 116 are mounted on semiconductor chips such as FPGAs and ASICs and operate in synchronization with the clock. In this embodiment, the clock frequency is 100 MHz.

[0038] Figure 4 is a flowchart showing the operation of the imaging process by the sample measuring device 100 of the first embodiment. The flowchart in Figure 4 is realized by the CPU 301 of the controller 116 executing a program. Figures 5(a), (b), and (c) illustrate the positional relationship of the sub-scanning of the array plate 101 during the imaging process, and show the array plate 101 viewed from above. Figures 6(a) and 6(b) illustrate the positional relationship of the height scanning of the array plate 101 during the imaging process, and are views of the array plate 101 from the side (long side). The position Z=0 on the vertical axis is the horizontal reference plane. The thickness and inclination of the placed array plate differ between Figure 6(a) and Figure 6(b).

[0039] In S401, the CPU 301 reads shooting conditions in response to a shooting instruction from the user. The user has previously input the shooting conditions via the user interface 117. The CPU 301 stores the acquired shooting conditions in the memory 303. Here, as the shooting conditions, the points 501(X1, Y1) and 502(X2, Y2) indicating the shooting area on the array plate 101, the pixel pitch Xp in the main scanning direction, the pixel pitch Yp in the sub-scanning direction, and the rotation speed Xs in the main scanning direction are input. In this embodiment, X1 = 500, X2 = 22500, Y1 = 500, Y2 = 64500, Xp = 10um, Yp = 10um, and Xs = 1200rpm. The memory 303 is paraphrased as a storage unit that stores information regarding the shooting area defined with respect to the surface (one surface) of the array plate 101 having a plurality of spots 202. The CPU 301 sets the shooting conditions in the synchronization circuit 311. Here, a rectangular area with the points 501 and 502 on the array plate 101 as diagonals is called the shooting area 503. Also, the CPU 301 calculates the number of pixels Nx = (X2 - X1) / Xp in the main scanning direction and the number of pixels Ny = (Y2 - Y1) / Yp in the sub-scanning direction in advance. In this embodiment, Nx = 2200 and Ny = 6400.

[0040] In S402, the CPU 301 acquires the height information of the array plate 101 and acquires tilt information based on the height information. The CPU 301 corresponds to an example of an acquisition unit. The tilt information corresponds to an example of information regarding the array plate. The height from the horizontal reference plane to the surface of the array plate 101, that is, the surface on the spot 202 side, is called the height information. Also, the tilt of the array plate 101 in the sub-scanning direction with respect to the horizontal reference plane is called the tilt information. Here, the CPU 301 acquires the height information Z3 and Z4 for two Y coordinates Y3 and Y4 by measurement. In this embodiment, Y3 = 750 and Y4 = 65000. In the case of the tilt as shown in Fig. 6(a), Z4 > Z3, and in the case of the tilt as shown in Fig. 6(b), Z3 < Z4. The method for acquiring the height information will be described later using the flowchart of Fig. 7. The tilt information K is calculated by the following (Equation 1). K = (Z4 - Z3) / (Y4 - Y3) ... (Equation 1)

[0041] In S403, the CPU301 calculates the target height from the reference plane at each sub-scan position (each row) according to the imaging conditions, height information, and tilt information K. Here, the target height Z(Y) at coordinate Y of any sub-scan position is given by the following (Equation 2). Z(Y) = K × (Y - Y³) + Z³ ... (Equation 2) In Figures 6(a) and 6(b), the target height 601 corresponds to the Z coordinate of the surface of the array plate 101.

[0042] In S404, the CPU 301 instructs the motor control circuits 307, 308, and 309 to move the array plate 101 and the light-emitting unit 104 to the shooting start position. In this embodiment, the X-coordinate of the shooting start position is the end of the scanning range of the piston crank mechanism 106, and the X-coordinate value is -2500. The Y-coordinate of the shooting start position is Y1 as specified in the shooting conditions. The Z-coordinate Z1 of the shooting start position is calculated as Z(Y1) in (Equation 1).

[0043] In S405, the CPU 301 starts the main scan, moving the light-emitting unit 104 in the main scanning direction (scanning process). Specifically, the CPU 301 instructs the motor control circuit 307 to rotate the pulse motor 107 at a rotational speed Xs. The rotation of the pulse motor 107 causes the light-emitting unit 104 to start reciprocating motion in the X direction. The encoder 108 and the coordinate calculation circuit 310 calculate the X coordinate of the light-emitting unit 104 at each clock cycle and output it to the synchronization circuit 311. In S406, the CPU 301 instructs the light source control circuit 305 to start emitting light from the light source 102. The light emitted from the light source 102 starts irradiating the array plate 101 with light via the light projector 104.

[0044] In S407, the synchronization circuit 311 determines whether the light-emitting unit 104 has reached the new line position. The synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104, output from the coordinate calculation circuit 310, moves from inside the imaging area to outside the imaging area. If the current main scanning direction is the forward direction (direction in which the X coordinate increases), the synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104 exceeds X2. The forward scanning is represented by the trajectory 504 in Figure 5(b). On the other hand, if the current main scanning direction is the return direction (direction in which the X coordinate decreases), the synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104 becomes smaller than X1. The return scanning is represented by the trajectory 506 in Figure 5(b). The initial value of the main scanning direction is the forward direction, and thereafter, each time the new line position is reached, the return direction and the forward direction are alternately repeated. If a newline position is reached, the synchronization circuit 311 outputs a sub-scan trigger signal and a height scan trigger signal, causing S416 and S417-S418 to operate in parallel. If a newline position is not reached, the synchronization circuit 311 proceeds to S408 without outputting a sub-scan trigger signal or a height scan trigger signal.

[0045] In S408, the synchronization circuit 311 determines whether the light-emitting unit 104 has reached the sampling position. The sampling position is the point on the array plate 101 where fluorescence signal data is acquired. Although the case where the sampling position is different from the spot is described below, it may also be the same as the spot. The X coordinate P(N) of the Nth sampling position is expressed by the following (Equation 3). P(N)=X1+Xp×(N+1 / 2) (N=0,1,…Nx-1) (Formula 3) The sampling positions are multiple points on the trajectory, such as point 508 in Figure 5(b), with a pitch of Xp in the X direction and a pitch of Yp in the Y direction. The initial value of the sampling position is P(0) and is stored inside the synchronization circuit 311. In determining the first sampling position, it is determined that the sampling position has been reached when the X coordinate of the light-emitting 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 sampling position from the second time onward, it is determined that the sampling position has been reached when the X coordinate of the light-emitting unit 104 output from the coordinate calculation circuit 310 passes through the sampling position updated in S410, which will be described later. If the sampling position is reached, the synchronization circuit 311 outputs a data acquisition trigger signal and proceeds to S409. If the sampling position has not yet been reached, the synchronization circuit 311 does not output a data acquisition trigger signal and proceeds to S411.

[0046] In S409, the data acquisition circuit 306 acquires optical information at the sampling position. Specifically, the data acquisition circuit 306 outputs a conversion start signal for its internal AD converter and performs AD conversion on the output voltage from the light sensor 105. The AD-converted fluorescence signal data is stored in the memory 303 via the DMA controller and memory control circuit 304 inside the data acquisition circuit 306. After storing Nx × Ny data points in the memory 303, the data acquisition circuit 306 sets its internal data acquisition completion register to 1, and sets it to 0 if it has not stored Nx × Ny data points in the memory 303. The processing in S409 is performed while the light-emitting unit 104 is moving back and forth in the main scanning direction, and not while it is moving in the sub-scanning direction.

[0047] In S410, the synchronization circuit 311 updates the sampling position it holds internally. The synchronization circuit 311 updates the sampling position P(N) to P(N+1) when the current main scanning direction is the forward path (direction in which the X coordinate increases), and updates the sampling position P(N) to P(N-1) when the current main scanning direction is the return path (direction in which the X coordinate decreases).

[0048] In S411, the CPU 301 determines whether data acquisition is complete or not. The CPU 301 reads the data acquisition completion register from the data acquisition circuit 306. If the value of the data acquisition completion register is 1, it determines that data acquisition is complete and proceeds to S412. If the value of the data acquisition completion register is 0, it determines that data acquisition is not complete and proceeds to S407.

[0049] In S412, the CPU 301 instructs the light source control circuit 305 to stop the light emission of the light source 102. When the light emission of the light source 102 stops, light irradiation to the array plate 101 via the light emitter 104 stops. In S413, the CPU 301 stops the main scan. Specifically, the CPU 301 instructs the motor control circuit 307 to stop the rotation of the pulse motor 107. When the rotation of the pulse motor 107 stops, the reciprocating motion of the light-emitting unit 104 in the X direction stops.

[0050] In S414, the CPU 301 instructs the motor control circuits 307, 308, and 309 to move the array plate 101 and the light-emitting unit 104 to the stop position. The X, Y, and Z coordinates of the stop position are 0. The movement to the stop position is performed by returning each axis to its home position using the Z-phase pulse signal from the encoder 108, the home sensor signal in the linear stage 109, and the home sensor signal from the linear stage 112.

[0051] In S415, the CPU 301 reads Nx × Ny fluorescence signal data stored in memory 303, performs data compression and format conversion processing, and creates a fluorescence image file in TIFF format. The fluorescence image file is stored in memory 303 and presented to the user via UI circuit 313 and user interface 117. Furthermore, upon instruction from the user, it can be transferred to an external data server via communication circuit 312.

[0052] In S416, the CPU 301 performs a sub-scan (scanning process) to move the array plate 101 in the sub-scan direction. Specifically, the CPU 301 instructs the motor control circuit 308 to move the array plate 101 by Yp in the sub-scan direction. Here, if My is the amount of movement of the array plate 101 when one voltage pulse signal is sent to the motor driver 111, then the number of pulses that the motor control circuit 308 outputs to the motor driver 111 is Yp / My. In this embodiment, My = 2um. The sub-scan is represented by trajectories 505 and 507 shown in Figure 5(b), and the distance moved in the Y direction is Yp. Figure 5(c) is an enlarged view of trajectories 505 and 507. Trajectory 505 includes a linear trajectory 511 along the forward direction of the main scanning direction, a semicircular trajectory 512, and a linear trajectory 513 along the return direction of the main scanning direction. Trajectory 507 includes a linear trajectory 514 along the return direction of the main scanning direction, a semicircular trajectory 515, and a linear trajectory 513 along the forward direction of the main scanning direction. Thus, in this embodiment, trajectories 505 and 507 each include trajectories corresponding to at least two or more directions of movement. Based on instructions 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-scan is completed while the light-emitting unit 104 is outside the imaging area in the main scanning direction.

[0053] In S417, the CPU 301 reads the target height before and after the sub-scan from the memory 303 in order to perform a height scan. Specifically, the CPU 301 reads Z(Y+Yp) and Z(Y) and calculates the amount of movement for the height scan. The amount of movement for the height scan varies depending on the current Y coordinate, but the CPU 301 calculates the number of output pulses and the rotation direction of the pulse motor 113 so that the height in the Z direction after the movement is as close as possible to the target height Z(Y+Yp) in the Y coordinate after the sub-scan in S416.

[0054] Let me explain the specific calculation method. Here, Mz is defined as the amount of movement of the array plate 101 when one voltage pulse signal is sent to the motor driver 114. In this embodiment, Mz = 1 µm. Let RoundMz(x) be the multiple of Mz closest to a certain number x, ABS(x) be the absolute value of a certain number, and Sign(x) be the sign of a certain number. The multiple of Mz closest to the target height is called the target pulse number. The target pulse number takes discrete values ​​and corresponds to a height of 602 in Figures 6(a) and 6(b). The number of pulses that the motor control circuit 309 outputs to the motor driver 114 is expressed by the following equation (4). Zp=ABS(RoundMz(Z(Y+Yp))-RoundMz(Z(Y))) ...(Formula 4) RoundMz(Z(Y)) is a discrete value close to the surface of the array plate 101, as shown by the height 602 in Figures 6(a) and 6(b).

[0055] Furthermore, the vertical movement direction Dir of the array plate is expressed by the following (Equation 5). Dir=Sign(RoundMz(Z(Y+Yp))-RoundMz(Z(Y))) ...(Formula 5) The positive direction for Dir is defined as vertically upward, and the direction in which the distance between the light-emitting unit 104 and the array plate 101 increases. When the inclination of the array plate 101 is as shown in Figure 6(a), the value of Dir is 1, and when it is as shown in Figure 6(b), the value of Dir is -1.

[0056] In S418, the CPU 301 performs a height scan to adjust the array plate 101 along the vertical direction in order to acquire focused optical information (adjustment step). Specifically, the CPU 301 instructs the motor control circuit 309 to move the array plate 101 by Zp in the direction of movement Dir. If the value of Dir is positive, the array plate 101 is moved upward, and if the value of Dir is negative, the array plate 101 is moved downward. The processing in S418 is performed based on the scan sequence information, which indicates that the light-emitting unit 104 has reached the new line position in S407. In other words, the CPU 301 decides whether or not to perform a height scan based on the scan sequence information. The motor control circuit 309 outputs a pulse count of Zp / Mz to the motor driver 111. Based on instructions 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 scan is completed while the light emitter 104 is outside the imaging area in the main scanning direction. Therefore, the height scan is performed during the sub-scan period. In other words, the height scan and sub-scan are performed in parallel. On the other hand, the height scan is not performed during the main scan period.

[0057] Once the sub-scan in S416 and the height scan in S418 are complete, proceed to S419. In S419, the synchronization circuit 311 updates the current main scanning direction and Y coordinate. Specifically, if the previous main scanning direction was the forward direction, the synchronization circuit 311 updates the main scanning direction to the return direction and sets the newline position to X1. On the other hand, if the previous main scanning direction was the return direction, the synchronization circuit 311 updates the main scanning direction to the forward direction and sets the newline position to X2. The synchronization circuit 311 also increments the current Y coordinate by Yp from the previous Y coordinate and proceeds to S411.

[0058] Figure 7 is a flowchart showing the operation for acquiring height information of the array plate 101, and corresponds to a part of the process S402 described above. In the flowchart of Figure 7, unlike the flowchart of Figure 4, the reflected light signal data is acquired and analyzed while performing height scanning at equal pitches without performing sub-scanning, and the surface height of the array plate 101 is calculated.

[0059] Figures 8(a) and (b) illustrate the positional relationship of the array plate 101 during height scanning in the height information acquisition operation, and are views of the array plate 101 from the side (short side). Figure 8(c) plots the amount of reflected light acquired by the optical sensor 105 during height scanning for each height, with the horizontal axis representing the magnitude of the light amount and the vertical axis representing the acquired height.

[0060] In S701, the CPU 301 sets parameters for acquiring height information in the synchronization circuit 311. The parameters include the Y coordinate Yh of the position where height information is acquired, the pixel pitch Xp in the main scanning direction, the pixel pitch Zp in the height scanning direction, points 801 (X5, Z5) and 802 (X6, Z6) indicating the height scanning range on the XZ plane, and the rotation speed Xs in the main scanning direction. The rectangular region 803, formed by points 801 and 802 on the XZ plane, is called the height scanning region. The CPU 301 calculates 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. In this embodiment, X5=500, X6=22500, Z5=2000, Z6=6000, Xp=10[um], Zp=10[um], and Xs=1200[rpm]. In this case, Nx=2200 and Nz=400.

[0061] In S702, the CPU 301 instructs the motor control circuits 307, 308, and 309 to move the array plate 101 and the light-emitting unit 104 to the starting position for acquiring height information. In this embodiment, the X-coordinate of the starting position for acquiring height information is the end of the scanning range of the piston crank mechanism 106, and the X-coordinate value is -2500. The Y-coordinate of the starting position for acquiring height information is Yh as specified by the parameter. The Z-coordinate of the starting position for acquiring height information is Z5 as specified by the parameter.

[0062] In S703, CPU301 starts the main scan. This process is the same as the process described in S405 above. In S704, the CPU 301 starts light irradiation by initiating the emission of light from the light source 102. This process is the same as the process described in S406 above.

[0063] In S705, the synchronization circuit 311 determines whether the light-emitting unit 104 has reached the new line position. The synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104, output from the coordinate calculation circuit 310, moves from inside the imaging area to outside the imaging area. If the current main scanning direction is the forward path direction (direction in which the X coordinate increases), the synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104 exceeds X6. The forward scanning is represented by the trajectory 804 in Figure 8(b). On the other hand, if the current main scanning direction is the return path direction (direction in which the X coordinate decreases), the synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104 becomes smaller than X5. The return scanning is represented by the trajectory 806 in Figure 8(b). The initial value of the main scanning direction is the forward path direction, and thereafter, each time the new line position is reached, the return and forward directions are alternated. If a newline position is reached, the synchronization circuit 311 outputs a height scan trigger signal and proceeds to S714. If a newline position has not been reached, the synchronization circuit 311 does not output a height scan trigger signal and proceeds to S706.

[0064] In S706, the synchronization circuit 311 determines whether the light-emitting unit 104 has reached the sampling position. This process is the same as the process in S408 described above. The sampling position is one of several points on the trajectory, such as point 808 in Figure 8(b), with a pitch of Xp in the X direction and a pitch of Zp in the Z direction. If the sampling position is reached, the synchronization circuit 311 outputs a data acquisition trigger signal and proceeds to S707. If the sampling position has not yet been reached, the synchronization circuit 311 does not output a data acquisition trigger signal and proceeds to S709.

[0065] In S707, the data acquisition circuit 306 acquires reflected light signal data from the sampling position. This process is the same as that described in S409. After storing Nx × Nz data points in memory 303, the data acquisition circuit 306 sets its internal data acquisition completion register to 1, and sets it to 0 if it has not stored Nx × Nz data points in memory 303.

[0066] In S708, the synchronization circuit 311 updates the sampling position it holds internally. This process is the same as the process described in S410 above. In S709, CPU301 determines whether data acquisition is complete or not. This process is the same as that described in S411. If it is determined that data acquisition is complete, the process proceeds to S710. If it is determined that data acquisition is not complete, the process proceeds to S705.

[0067] In S710, CPU301 stops the light emission from light source 102. This process is the same as the process in S412 described above. In S711, CPU301 stops the main scan. This process is the same as the process described in S413 above. In S712, the CPU 301 moves the array plate 101 and the light-emitting unit 104 to the stop position. This process is the same as the process in S414 described above.

[0068] In S713, the CPU 301 reads and analyzes Nx × Nz reflected light signal data stored in memory 303 to calculate the height information of the array plate. Specifically, it averages the Nx data acquired at the same height to determine the average light intensity for each height. When the average light intensity is arranged by Z coordinate, there are two peaks corresponding to the front and back surfaces of the array plate 101. In Figure 8(c), peak 809 shows the peak due to reflected light from the front surface of the array plate 101, and peak 810 shows the peak due to reflected light from the back surface of the array plate 101. Of the two peaks, peak 809, which has a larger Z coordinate, i.e., peak 811, which corresponds to the front surface, is the height information corresponding to position Yh.

[0069] In S714, the CPU 301 instructs the motor control circuit 309 to move the array plate 101 by Zp in the height scanning direction. If Mz is the amount of movement of the array plate 101 when one voltage pulse signal is sent to the motor driver 114, then the number of pulses that the motor control circuit 309 outputs to the motor driver 114 is Zp / Mz. The height scanning is represented by trajectories 805 and 807 shown in Figure 8(b), and the distance moved in the Z direction is Zp. Based on instructions 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 scan is completed while the light-emitting unit 104 is outside the imaging area in the main scanning direction.

[0070] In S715, the synchronization circuit 311 updates the current main scanning direction and Z coordinate. Specifically, if the previous main scanning direction was the forward direction, the synchronization circuit 311 updates the main scanning direction to the return direction and sets the newline position to X5. On the other hand, if the previous main scanning direction was the return direction, the synchronization circuit 311 updates the main scanning direction to the forward direction and sets the newline position to X6. The synchronization circuit 311 also increments the current Z coordinate by Zp from the previous Z coordinate and proceeds to S709.

[0071] In S402 described above, when acquiring tilt information, the flowchart in Figure 7 is processed for Y coordinates Y3 and Y4 respectively to obtain height information Z3 for Y coordinate Y3 and height information Z4 for Y coordinate Y4, and tilt information K is calculated using (Equation 1). By obtaining the tilt information K in this way, the target height of each sub-scan position can be calculated in S403.

[0072] In the sample measuring device 100 of this embodiment, a height scan is performed during the sub-scanning period to adjust the array plate 101 along the vertical direction. Therefore, focused optical information can be acquired in a short time. Furthermore, in this embodiment, by using the synchronization circuit 311, sub-scanning and height scanning are performed in synchronization with the position of the light-emitting unit 104. By performing sub-scanning and height scanning while the light-emitting unit 104 is outside the imaging area, it is not necessary to move the array plate 101 within the imaging area. Therefore, the effects of vibrations associated with the driving of the array plate 101 can be reduced. In addition, since the sample measurement device 100 does not have a high-speed servo control system consisting of a high-performance focus sensor or a low-vibration actuator, a fluorescence image that is in focus across the entire array plate 101 can be acquired with a simple configuration.

[0073] Furthermore, in this embodiment, the pulse motor 107 for main scanning rotates at a constant speed during imaging to move the light-emitting unit 104, and sub-scanning and height scanning are performed when the light-emitting unit 104 is outside the imaging area. Since it is not necessary to temporarily stop the pulse motor 107 for main scanning before sub-scanning or height scanning, the light-emitting unit 104 can be scanned back and forth at high speed, and the imaging time can be shortened.

[0074] Furthermore, in this embodiment, two-dimensional scanning of the array plate 101 is performed, with a main scan and a sub-scan, while correcting for individual differences in the thickness and inclination of the entire array plate 101 based on height information and inclination information acquired in advance at at least two points. Therefore, compared to scanning the array plate 101 in three dimensions, a fluorescence image that is in focus across the entire array plate 101 can be obtained in a short time.

[0075] Furthermore, in this embodiment, the target pulse count closest to the target height is calculated for each sub-scan position (each row). Therefore, even if the amount and direction of height scanning are not predetermined values, especially if the amount of movement differs from row to row, it is possible to adjust to a height close to the target height. Consequently, even if there are individual differences in the thickness and inclination of the array plate 101 and the method of mounting the array plate 101, and it is difficult to predict the height and inclination of the array plate 101 in advance, it is possible to obtain a fluorescence image that is in focus across the entire surface.

[0076] Furthermore, in this embodiment, when acquiring height information, a main scan is also performed and Nx data points are averaged, allowing for stable peak detection even if there is partial dirt or liquid on the array plate 101. Therefore, the accuracy of the acquired height information can be improved compared to detecting a peak at a single point on the array plate 101 for each height scan position.

[0077] Furthermore, in this embodiment, by using the synchronization circuit 311, fluorescence signal data sampling is performed in synchronization with the position of the light-emitting unit 104. Compared to the case where data sampling or sub-scanning is performed at a fixed period without synchronization with the position of the light-emitting unit 104, it is possible to acquire equally spaced data across the entire array plate 101, thereby improving the accuracy of the position when acquiring captured images.

[0078] In this embodiment, the imaging area 503 covers the entire area 204, and the case where all spots on the array plate 101 are imaged has been described, but this is not the only case. For example, the user can set any part of area 204 as the imaging area 503. In this case, the imaging time can be shortened by scanning only the portion containing the spots of interest to the user.

[0079] In this embodiment, the case in which height information is obtained using the peak of reflected light from the surface of the slide glass 201 has been described, but this is not the only case. For example, height information may be obtained using the peak position of the luminance of the fluorescence signal from some spots on the array plate 101. In this case, it is necessary to illuminate some spots with light in order to obtain height information, but it is not necessary to obtain reflected light with the light sensor 105, so the number of components in the optical system can be reduced.

[0080] In this embodiment, the case where there is only one wavelength from the light source 102 has been described. However, it is also possible to provide multiple light sources, optical systems, and photosensors for each wavelength, and irradiate the array plate 101 with excitation light of multiple wavelengths. By comparing the fluorescence signals generated from excitation light of multiple wavelengths, the properties of the biological material on the spot can be analyzed in more detail.

[0081] In this embodiment, the case in which the light-emitting unit 104 is moved in the main scanning direction has been described, but this is not the only case. For example, the array plate 101 may be moved in the main scanning direction, or both the light-emitting unit 104 and the array plate 101 may be moved in the main scanning direction. In other words, a configuration in which at least one of the light-emitting unit 104 and the array plate 101 is moved relative to the main scanning direction is also possible. Furthermore, although this embodiment describes the case in which the array plate 101 is moved in the sub-scanning direction, it is not limited to this case. For example, the light-emitting unit 104 may be moved in the sub-scanning direction, or both the light-emitting unit 104 and the array plate 101 may be moved in the sub-scanning direction. In other words, it is also possible to configure it so that at least one of the light-emitting unit 104 and the array plate 101 is moved relative to the other in the sub-scanning direction. Furthermore, although this embodiment describes the case in which the array plate 101 is moved in the vertical direction, it is not limited to this case. For example, the light-emitting unit 104 may be moved in the vertical direction, or both the light-emitting unit 104 and the array plate 101 may be moved in the vertical direction. In other words, the relative position between the light-emitting unit 104 and the array plate 101 may be adjusted by relatively moving at least one of the light-emitting unit 104 and the array plate 101 in the vertical direction.

[0082] (Second embodiment) The second embodiment differs from the first embodiment in the scanning method of the light-emitting unit 104 and the sampling method of the fluorescence signal data. In the first embodiment, signals from the optical sensor 105 were acquired in the forward and return directions of the main scan, and sub-scanning and height scanning were performed while the ends of the array plate 101 were outside the imaging area. In this embodiment, signals from the optical sensor 105 are acquired in the forward direction of the main scan, and sub-scanning and height scanning are performed in the return direction of the main scan. The configuration of the sample measuring device 100, the array plate 101, and the internal configuration of the controller 116 in this embodiment are the same as in Figures 1, 2, and 3, and therefore their explanation will be omitted.

[0083] Figure 9 is a diagram illustrating the positional relationship of the sub-scanning of the array plate 101 during the imaging process. Figure 10 is a flowchart showing the operation of the imaging process of the array plate 101 in the sample measuring device 100 of this embodiment.

[0084] Steps S1001 to S1006 are the same as steps S401 to S406 in the first embodiment. In S1007, the synchronization circuit 311 determines whether the light-emitting unit 104 has reached the new line position. The synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104, output from the coordinate calculation circuit 310, moves from inside the imaging area to outside the imaging area. If the current main scanning direction is the forward direction (direction in which the X coordinate increases), the new line position is determined to have been reached when the X coordinate of the light-emitting unit 104 exceeds X2. The forward scanning is represented by the trajectory 901 in Figure 9. On the other hand, if the current main scanning direction is the return direction (direction in which the X coordinate decreases), the synchronization circuit 311 determines that the new line position has been reached when the X coordinate of the light-emitting unit 104 becomes smaller than X1. The return scanning is represented by the trajectory 902 in Figure 9. The initial value of the main scanning direction is the forward direction, and thereafter, each time the new line position is reached, the return direction and the forward direction are alternately repeated. If a line break is reached, proceed to S1017. If a line break is not reached, proceed to S1008.

[0085] In S1008, the synchronization circuit 311 determines whether the current main scanning direction is the forward or return path. If it is the forward path, proceed to S1009. If it is the return path, proceed to S1012. In S1009, the synchronization circuit 311 determines whether the light-emitting unit 104 has reached a sampling position. A sampling position is a point on the array plate 101 where fluorescence signal data is acquired. The X coordinate P(N) of the Nth sampling position is expressed by (Equation 3) described above. The sampling positions are multiple points on the trajectory, such as point 903 in Figure 9, with a pitch of Xp in the X direction and a pitch of Yp in the Y direction, and the coordinates are the same as in the first embodiment. The initial value of the sampling position is P(0) and is stored inside the synchronization circuit 311. In determining the first sampling position, it is determined that the sampling position has been reached when the X coordinate of the light-emitting 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 determining the second and subsequent sampling positions, it is determined that the sampling position has been reached when the X coordinate of the light-emitting unit 104 output from the coordinate calculation circuit 310 passes the sampling position updated in S1011 (described later) in the forward direction. If the sampling position is reached, the synchronization circuit 311 outputs a data acquisition trigger signal and proceeds to S1010. If the sampling position has not yet been reached, the synchronization circuit 311 does not output a data acquisition trigger signal and proceeds to S1012.

[0086] S1010 is the same as the process in S409 of the first embodiment. In S1011, the synchronization circuit 311 updates the sampling position it holds internally. The synchronization circuit 311 updates the sampling position P(N) to P(N+1) because the current main scanning direction is the forward path direction (the direction in which the X coordinate increases). However, if N = Nx-1, it updates P(N) to P(0).

[0087] Steps S1012 to S1016 are the same as those in steps S401 to S406 of the first embodiment. In S1017, the synchronization circuit 311 determines whether the current main scanning direction is the forward or return path. If it is the forward path, the synchronization circuit 311 outputs a sub-scan trigger signal and a height scan trigger signal, and operates S1018 and S1019-S1020 in parallel. If it is the return path, proceed to S1021.

[0088] In S1018, the CPU 301 performs a sub-scan simultaneously with the main scan in the return direction. Specifically, the CPU 301 instructs the motor control circuit 308 to move the array plate 101 by Yp in the sub-scan direction. Here, if My is the amount the array plate 101 moves when one voltage pulse signal is sent to the motor driver 111, then the number of pulses that the motor control circuit 308 outputs to the motor driver 111 is Yp / My. In this embodiment, My = 2um. The sub-scan here is performed simultaneously with the main scan in the return direction, resulting in a sloping, approximately straight trajectory that intersects both the X and Y directions, as shown in trajectory 902 in Figure 9. The distance traveled in the Y direction is Yp. Based on instructions 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-scan is completed while the light-emitting unit 104 is moving in the return path direction of the main scanning direction.

[0089] S1019 is the same as the process in S417 of the first embodiment. In S1020, height scanning is performed to acquire focused optical information. Specifically, the CPU 301 instructs the motor control circuit 309 to move the array plate 101 by Zp in the direction of movement Dir. If the value of Dir is positive, the array plate 101 is moved upward, and if the value of Dir is negative, the array plate 101 is moved downward. The motor control circuit 309 outputs a pulse of Zp / Mz to the motor driver 111. Based on instructions 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 scan is completed while the light emitter 104 is moving in the return direction of the main scan. Therefore, the height scan is performed while the sub-scan is being performed and while the main scan in the return direction is being performed. In other words, the height scan and the sub-scan and main scan in the return direction are performed in parallel. On the other hand, the height scan is not performed while the main scan in the forward direction is being performed.

[0090] Once the sub-scan in S1018 and the height scan in S1020 are complete, proceed to S1021. S1021 is the same as the process in S419 of the first embodiment.

[0091] In the sample measuring device 100 of this embodiment, fluorescence signal data is acquired only in the forward direction of the main scan, and sub-scanning and height scanning are performed in the return direction of the main scan without acquiring fluorescence signal data. Therefore, although the time required for imaging is doubled, the influence of positional and angular errors of the light-emitting unit 104 due to the forward and return paths can be reduced because the main scan direction when acquiring data can be made consistent.

[0092] Furthermore, in this embodiment, the time spent on the return path of the main scan can be allocated to the sub-scan and height scan, allowing the speeds of the sub-scan and height scan to be reduced. Consequently, the influence of residual vibrations caused by the sub-scan and height scan can be reduced, thereby improving the image quality of the fluorescence image.

[0093] In this embodiment, the case in which fluorescence signal data is acquired only in the forward direction of the main scan has been described. However, it is also possible to acquire fluorescence signal data only in the return direction of the main scan, and perform sub-scanning and height scanning without acquiring fluorescence signal data in the forward direction of the main scan. In this case, it can be achieved by reversing the forward and return directions in the determinations in S1008 and S1017 described above.

[0094] In this embodiment, the operation of the imaging process has been described, but similarly, in the height information acquisition process, sampling and height scanning can be performed in either the forward or return direction only. In this case, as with the imaging process, the influence of positional and angular errors of the light-emitting unit 104 can be reduced, and the influence of residual vibrations during height scanning can be minimized, thereby improving the accuracy of the acquired height information.

[0095] (Third embodiment) The third embodiment differs from the first embodiment in that it does not have an encoder 108 for measuring the position of the light-emitting unit 104 in the main scanning direction. In the first embodiment, the coordinate calculation circuit 310 calculated the position of the light-emitting unit 104 based on the signal from the encoder 108. In this embodiment, the position of the light-emitting unit 104 is calculated based on the motor drive pulse signal from the motor control circuit 307. Note that the configuration of the sample measuring device 100 and the array plate 101 in this embodiment are the same as in Figures 1 and 2, so their description is omitted.

[0096] Figure 11 is a block diagram showing the internal configuration of the controller 1116 of the third embodiment. The controller 1116 has the same configuration as the first embodiment, except that the coordinate calculation circuit 310 is replaced with a coordinate calculation circuit 1310 and it does not have an encoder 108. The coordinate calculation circuit 1310 is a circuit that calculates the position of the light-emitting unit 104 based on the drive pulse voltage from the motor control circuit 307.

[0097] Figure 12 is a diagram illustrating the operation of the piston-crank mechanism. If the length of the crank 118 in the piston-crank mechanism is r, the length of the connecting rod 119 is l, and the angle of the pulse motor 107 is θ, then the position x of the light-emitting unit 104 is expressed by the following equation (Equation 6).

[0098]

number

[0099] θ can be calculated by multiplying the rotation angle per pulse by the number of drive pulses. Since r and l are known values, the coordinates of the light-emitting unit 104 are calculated using (Equation 6) each time a drive pulse is input. However, since the rotation angle per pulse is in increments of 0.72°, the obtained x values ​​are discrete. Therefore, the coordinate calculation circuit 1310 estimates the coordinates between pulses by interpolating them using the angular velocity.

[0100] Figure 13 is a flowchart showing the operation of the coordinate calculation circuit. In S1301, the coordinate calculation circuit 1310 sets the values ​​of its internal pulse counter and time counter to 0. In S1302, the coordinate calculation circuit 1310 determines whether or not the rising edge of the drive pulse signal from the motor control circuit 307 has been input. If it has been input, the process proceeds to S1303. If it has not been input, the process proceeds to S1311.

[0101] In S1303, the coordinate calculation circuit 1310 determines whether the motor has completed one rotation. For example, if the rotation angle θp of the pulse motor 107 per pulse is 0.72°, the motor completes one rotation in 500 pulses. Therefore, if the current value Cp of the pulse counter is 499, it is determined that one rotation has occurred and the process proceeds to S1310. On the other hand, if the current value Cp of the pulse counter is 498 or less, it is determined that one rotation has not occurred and the process proceeds to S1304.

[0102] In S1304, the coordinate calculation circuit 1310 increments the pulse counter by 1. In S1305, the coordinate calculation circuit 1310 calculates the angular velocity w by dividing the value of the time counter by the clock period. If the value of the time counter is Ct and the clock period is T, the angular velocity w is calculated as w = θp × T / Ct. However, if the value of the time counter Ct is 0, the angular velocity w is calculated to be 0.

[0103] In S1306, the coordinate calculation circuit 1310 sets the time counter to 0. In S1307, the coordinate calculation circuit 1310 calculates the angle θ. Here, if the value of the pulse counter is Cp, the angle θ is calculated as θ = Cp × θp + w × Ct × T.

[0104] In S1308, the coordinate calculation circuit 1310 calculates the x-coordinate. Specifically, the x-coordinate is calculated by substituting the calculated angle θ into (Equation 6). In S1309, the coordinate calculation circuit 1310 outputs the calculated coordinates to the synchronization circuit 311.

[0105] In S1310, the coordinate calculation circuit 1310 sets the pulse counter to 0. In S1311, the coordinate calculation circuit 1310 increments the pulse counter by 1. As described above, the operation of the coordinate calculation circuit 1310 has been explained using a flowchart. However, since the coordinate calculation circuit 1310 is implemented on a digital circuit, it actually performs operations S1301 to S1311 for each clock cycle.

[0106] In this embodiment, by estimating the position of the light-emitting unit 104 from the drive pulse signal of the motor control circuit 307, it is possible to synchronize the main scan, sub-scan, height scan, and data acquisition without using an encoder. Since the sample measuring device 100 does not have an encoder, manufacturing costs can be reduced.

[0107] In this embodiment, the rotation angle θp of the pulse motor 107 per pulse was set to 0.72°, but this is not the only case. For example, by using a motor driver with a microstepping control function in the motor control circuit 307, θp can be divided into tens to hundreds of parts, thereby improving the accuracy of estimating the position of the light-emitting unit 104.

[0108] In this embodiment, the coordinate calculation circuit 1310 calculates the angular velocity from the time difference of the rising edge of the pulse signal, but this is not the only case. For example, the coordinate calculation circuit 1310 may be operated while the pulse motor 107 is rotating at a constant speed, and the angular velocity may be calculated using the rotation speed Xs of the main scanning direction specified by the user.

[0109] Although the present invention has been described in detail above based on its preferred embodiments, the present invention is not limited to any particular embodiment, and various forms that do not depart from the spirit of the invention are also included in the present invention. For example, parts of the configuration or process of each embodiment may be combined with other embodiments. Furthermore, the various controls described above, which are performed by the CPU 301, synchronization circuit 311, data acquisition circuit 306, etc., in the above-described embodiment, may be performed by a single piece of hardware. Alternatively, the various controls described above may be shared among multiple pieces of hardware (for example, multiple processors or circuits) to control the entire device.

[0110] <Other Embodiments> The present invention can also be realized by performing the following process: supplying a program that implements the functions of the above-described embodiment to a system or device via a network or various recording media, and having the computer (CPU, MPU, etc.) of that system or device read and execute the program code. In this case, the program and the recording media storing the program constitute the present invention.

[0111] Furthermore, the disclosure of this embodiment includes the following configuration. (Composition 1) A scanning device for scanning an observation optical system over an array plate having multiple spots on one side, An observation optical system that irradiates one of the surfaces with primary light in order to acquire optical information relating to at least a portion of the plurality of spots, A scanning unit that performs a main scan in which the observation optical system moves relative to the array plate in a first direction and acquires the optical information, and a sub-scan in which the observation optical system moves relative to the array plate in a second direction intersecting the first direction without acquiring the optical information. It includes an adjustment unit for adjusting the relative position of the observation optical system with respect to the array plate in the optical axis direction of the primary light, The scanning apparatus is characterized in that the adjustment unit performs the adjustment when the scanning unit is in the sub-scanning period. (Configuration 2) The scanning apparatus according to configuration 1, characterized in that the adjustment unit does not perform the adjustment when the scanning unit is in the period of the main scan. (Composition 3) The scanning device according to configuration 1 or 2, further comprising an image acquisition unit that acquires a two-dimensional image based on an output signal from the observation optical system and information regarding the relative position of the observation optical system with respect to the array plate in a plane in which the observation optical system moves relative to the array plate. (Composition 4) A scanning device according to any one of configurations 1 to 3, further comprising a control unit that determines whether or not to perform the adjustment based on information regarding the scanning sequence by the scanning unit. (Composition 5) A scanning device according to any one of configurations 1 to 4, further comprising a storage unit for storing information relating to a shooting area defined for one of the aforementioned surfaces. (Composition 6) The scanning apparatus according to any one of configurations 1 to 5, characterized in that the sub-scanning includes movement corresponding to two or more movement directions in a plane in which the observation optical system moves relative to the array plate. (Composition 7) The scanning device according to configuration 6, characterized in that the sub-scanning period includes movement in the first direction. (Composition 8) It has an acquisition unit that acquires information about the aforementioned array plate, The scanning device according to any one of configurations 1 to 7, characterized in that the adjustment unit performs the adjustment based on the information regarding the array plate acquired by the acquisition unit. (Composition 9) The scanning device according to configuration 8, characterized in that the information relating to the array plate includes information on the inclination of the array plate as viewed from the first direction. (Composition 10) The scanning device according to configuration 9, characterized in that the inclination information of the array plate is calculated based on height information acquired at least two points on the array plate. (Composition 11) The scanning device according to any one of configurations 1 to 10, characterized in that the scanning unit moves the observation optical system and the array plate relative to each other based on information regarding the position of the observation optical system and information regarding the position of the array plate. (Composition 12) The scanning device according to any one of configurations 1 to 11, characterized in that the adjustment unit performs the adjustment based on information relating to the imaging area from which the optical information is acquired and information relating to the position of the observation optical system. (Composition 13) The scanning apparatus according to configuration 12, characterized in that the adjustment unit performs the adjustment when the position of the observation optical system is outside the imaging area. (Composition 14) The scanning device according to configuration 12 or 13, characterized in that the information relating to the imaging area is information that has been previously entered by the user. (Composition 15) It has a measuring unit that measures the position of the observation optical system, The scanning device according to any one of configurations 11 to 14, characterized in that the information relating to the position of the observation optical system is acquired based on the position of the observation optical system measured by the measuring unit. (Composition 16) The observation optical system has a drive unit that moves it relative to the array plate, The scanning device according to any one of configurations 11 to 14, characterized in that the information relating to the position of the observation optical system is acquired based on a signal that drives the drive unit. (Composition 17) The scanning device according to any one of configurations 1 to 16, characterized in that the second direction is a direction orthogonal to the first direction. (Composition 18) The scanning device according to any one of configurations 1 to 16, characterized in that the second direction is not perpendicular to the first direction and is inclined with respect to the first direction. (Composition 19) The array plate, when viewed from one of the sides, has a rectangular shape with a short side and a long side, The first direction is parallel to the short side of the array plate, The scanning device according to any one of configurations 1 to 16, characterized in that the second direction is parallel to the long side of the array plate. (Composition 20) The array plate, when viewed from one of the sides, has a rectangular shape with a short side and a long side, The first direction is parallel to the short side of the array plate, The scanning device according to any one of configurations 1 to 16, characterized in that the second direction intersects with both the short side and the long side of the array plate. (Composition 21) The scanning device according to any one of configurations 1 to 20, characterized in that the observation optical system is configured to irradiate at least a portion of the plurality of spots with primary light and to collect secondary light from at least a portion of the plurality of spots. (Method 1) A scanning method for scanning an observation optical system over an array plate having multiple spots on one side, A scanning process comprising: a main scan in which an observation optical system that irradiates primary light toward one surface in order to acquire optical information relating to at least a portion of the plurality of spots is moved relative to the array plate in a first direction and the optical information is acquired; and a sub-scan in which the observation optical system is moved relative to the array plate in a second direction intersecting the first direction without acquiring the optical information. The process includes an adjustment step for adjusting the relative position of the observation optical system with respect to the array plate in the optical axis direction of the primary light, The scanning method is characterized in that the adjustment step is performed during the sub-scanning period of the scanning step. (Method 2) The process includes an acquisition step that acquires information about the array plate in advance, prior to the scanning step. The scanning method according to Method 1, characterized in that the adjustment step is performed based on the information regarding the array plate acquired in the acquisition step. (Program 1) A program that causes a computer to perform each step described in Method 1. [Explanation of Symbols]

[0112] 100: Sample measuring device (scanning device) 101: Array plate 104: Light emitter 105: Light sensor 106: Piston crank mechanism 107: Pulse motor 109: Linear stage 110: Pulse motor 112: Linear stage 113: Pulse motor

Claims

1. A scanning device for scanning an observation optical system over an array plate having multiple spots on one side, An observation optical system that irradiates one of the surfaces with primary light in order to acquire optical information relating to at least a portion of the plurality of spots, A scanning unit that performs a main scan in which the observation optical system moves relative to the array plate in a first direction and acquires the optical information, and a sub-scan in which the observation optical system moves relative to the array plate in a second direction intersecting the first direction without acquiring the optical information. It includes an adjustment unit for adjusting the relative position of the observation optical system with respect to the array plate in the optical axis direction of the primary light, The scanning apparatus is characterized in that the adjustment unit performs the adjustment based on information relating to the imaging area for acquiring the optical information and information relating to the position of the observation optical system when the position of the observation optical system is outside the imaging area and the scanning unit is in the sub-scanning period.

2. The scanning apparatus according to claim 1, characterized in that the adjustment unit does not perform the adjustment when the scanning unit is in the period of the main scan.

3. The scanning apparatus according to claim 1, further comprising an image acquisition unit that acquires a two-dimensional image based on an output signal from the observation optical system and information relating to the relative position of the observation optical system with respect to the array plate in a plane in which the observation optical system moves relative to the array plate.

4. The scanning apparatus according to claim 1, further comprising a control unit that determines whether or not to perform the adjustment based on information regarding the scanning sequence by the scanning unit.

5. The scanning device according to claim 1, further comprising a storage unit for storing information relating to a shooting area defined for one of the aforementioned surfaces.

6. The scanning apparatus according to claim 1, characterized in that the sub-scanning includes movement corresponding to two or more movement directions in a plane in which the observation optical system moves relative to the array plate.

7. The scanning apparatus according to claim 6, characterized in that the sub-scanning includes movement in the first direction.

8. It has an acquisition unit that acquires information about the aforementioned array plate, The scanning apparatus according to claim 1 or 2, characterized in that the adjustment unit performs the adjustment based on the information regarding the array plate acquired by the acquisition unit.

9. The scanning device according to claim 8, characterized in that the information relating to the array plate includes information on the inclination of the array plate as viewed from the first direction.

10. The scanning device according to claim 8, characterized in that the inclination information of the array plate is calculated based on height information obtained at least two points on the array plate.

11. The scanning device according to claim 1, characterized in that the scanning unit moves the observation optical system and the array plate relative to each other based on information regarding the position of the observation optical system and information regarding the position of the array plate.

12. The scanning device according to claim 11, characterized in that the information regarding the imaging area is information that has been previously entered by the user.

13. It has a measuring unit that measures the position of the observation optical system, The scanning device according to claim 11 or 12, characterized in that the information relating to the position of the observation optical system is information obtained based on the position of the observation optical system measured by the measuring unit.

14. The observation optical system has a drive unit that moves it relative to the array plate, The scanning device according to claim 11 or 12, characterized in that the information relating to the position of the observation optical system is information acquired based on a signal that drives the drive unit.

15. The scanning device according to claim 1 or 2, characterized in that the second direction is perpendicular to the first direction.

16. The scanning device according to claim 1 or 2, characterized in that the second direction is not perpendicular to the first direction, but inclined with respect to the first direction.

17. The array plate, when viewed from one of the sides, has a rectangular shape with a short side and a long side, The first direction is parallel to the short side of the array plate, The scanning device according to claim 1 or 2, characterized in that the second direction is parallel to the long side of the array plate.

18. The array plate, when viewed from one of the sides, has a rectangular shape with a short side and a long side, The first direction is parallel to the short side of the array plate, The scanning device according to claim 1 or 2, characterized in that the second direction intersects with both the short side and the long side of the array plate.

19. The scanning apparatus according to claim 1 or 2, characterized in that the observation optical system is configured to irradiate at least a portion of the plurality of spots with primary light and to collect secondary light from at least a portion of the plurality of spots.

20. A scanning method for scanning an observation optical system over an array plate having multiple spots on one side, A scanning process comprising: a main scan in which an observation optical system that irradiates primary light toward one surface in order to acquire optical information relating to at least a portion of the plurality of spots is moved relative to the array plate in a first direction and the optical information is acquired; and a sub-scan in which the observation optical system is moved relative to the array plate in a second direction intersecting the first direction without acquiring the optical information. The process includes an adjustment step for adjusting the relative position of the observation optical system with respect to the array plate in the optical axis direction of the primary light, The scanning method is characterized in that, in the adjustment step, the adjustment is performed based on information regarding the imaging area from which the optical information is acquired and information regarding the position of the observation optical system, when the position of the observation optical system is outside the imaging area and during the sub-scan period of the scanning step.

21. The process includes an acquisition step that acquires information about the array plate in advance, prior to the scanning step. The scanning method according to claim 20, characterized in that the adjustment step is performed based on the information regarding the array plate acquired in the acquisition step.

22. A program for causing a computer to perform each of the steps described in claim 20.

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