Scanning device, scanning method, and program

Abstract

The scanning device according to the present invention comprises: an observation optical system that radiates light once toward one surface so as to acquire optical information related to at least some of a plurality of spots; a scanning unit that performs a main scan in which the observation optical system moves in a first direction with respect to the array plate 101 and acquires optical information, and a sub-scan in which the observation optical system moves in a second direction intersecting the first direction with respect to the array plate 101 and does not acquire optical information; and an adjustment unit that adjusts the position of the observation optical system with respect to the array plate 101 in the optical axis direction of the primary light. The adjustment unit performs adjustment in a case where the scanning unit is in a period of sub-scanning.

Description

Technical Field

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

[0002] A protein array plate or a peptide array plate is known in which a large number of biomolecules such as proteins and peptides having peptide bonds are fixed on a substrate. By using this protein array plate or peptide array plate, interactions with a large number of biomolecules fixed on the substrate can be performed simultaneously. Such an array plate is effective for comprehensively analyzing the interactions between a bio-derived liquid sample (e.g., blood, cell extract, saliva, or tissue fluid) and a large number of proteins or peptides. Through such an analysis, the characteristics of the sample can be measured.

[0003] The fixation sites of proteins or peptides on the substrate are called spots. As a method for observing the spots that interact with a sample, for example, a method for identifying which spot has interacted by labeling the spot with a fluorescent probe is known. A microarray scanner is known as a device for observing an array plate labeled with a fluorescent probe.

[0004] The specification of U.S. Patent No. 7911670 discusses a microarray scanner including an illumination optical system, a fluorescence detection optical system, and a two-dimensional scanning system. The illumination optical system has a function of focusing and irradiating a laser onto the array plate. The fluorescence detection optical system has a function of detecting the fluorescence intensity from the spots labeled with a fluorescent probe. The two-dimensional scanning system has a function of acquiring a fluorescence image of the spots on the array plate by performing two-dimensional scanning of the array plate or the optical system. In addition, a confocal optical system is used as the fluorescence detection optical system.

[0005] Patent No. 5281756 discusses a scanning optical device that simplifies the position adjustment in the height direction during fluorescence image acquisition.

[0006] Citation List

[0007] Patent Documents

[0008] PTL 1: U.S. Patent No. 7911670

[0009] PTL 2: Patent No. 5281756 Summary of the Invention

[0010] Technical Problem

[0011] Due to individual differences in the glass thickness and tilt of the array plate, a confocal optical system with a shallow depth of field has a problem that it is difficult to obtain a focused fluorescence image across the entire surface of the array plate.

[0012] In the specification of U.S. Patent No. 7911670, during two-dimensional scanning, automatic focus adjustment using a focus sensor is performed to obtain a focused fluorescence image across the entire surface of the array plate. However, in order to perform automatic focus adjustment with high precision while performing high-speed two-dimensional scanning of the array plate or the optical system, a high-speed feedback control system composed of a high-performance focus sensor and a low-vibration actuator is required, which increases the complexity of the device.

[0013] In Patent No. 5281756, in order to obtain a focused fluorescence image across the entire surface of the array plate, it is necessary to repeatedly perform two-dimensional scanning while changing the focus position and setting parameters, which increases the fluorescence measurement time.

[0014] The present invention has been made in view of the above problems, and the object of the present invention is to obtain focused optical information in a short time.

[0015] A scanning device configured to scan an observation optical system above an array plate, the array plate including a plurality of spots on one surface, the scanning device including: an observation optical system configured to radiate light once toward the one surface to obtain optical information related to at least a part of the plurality of spots; a scanning unit configured to perform main scanning and sub-scanning, in the main scanning, the observation optical system moves relative to the array plate in a first direction and obtains the optical information, in the sub-scanning, the observation optical system moves relative to the array plate in a second direction intersecting the first direction without obtaining the optical information; and an adjustment unit configured to adjust the position of the observation optical system relative to the array plate in the optical axis direction of the light, wherein the adjustment unit performs the adjustment when the scanning unit is in the period of the sub-scanning.

[0016] Advantageous Effects of the Present Invention

[0017] The present invention enables focused optical information to be obtained in a short time. Description of the Drawings

[0018] Figure 1 Figure 1 is a schematic diagram illustrating the configuration of a sample measurement device according to the first embodiment.

[0019] Figure 2A Figure 2A is an XY plan view illustrating the configuration of the array plate.

[0020] Figure 2B Figure 2B is a cross-sectional view illustrating the configuration of the array plate.

[0021] ​​​​​​​Figure 3 Figure 3 It is a diagram showing the internal configuration of the controller according to the first embodiment.

[0022] Figure 4 Figure 4 It is a flowchart showing the operation of the scanning device according to the first embodiment.

[0023] Figure 5A Figure 5A It is a diagram showing the scanning area of the scanning device on the array board according to the first embodiment.

[0024] Figure 5B Figure 5B It is a diagram showing the sub-scanning trajectory of the scanning device according to the first embodiment.

[0025] Figure 5C Figure 5C It is a partially enlarged view showing the sub-scanning trajectory of the scanning device according to the first embodiment.

[0026] Figure 6A Figure 6A It is a diagram showing an example of the height distribution of the array substrate detected by the scanning device in the sub-scanning direction according to the first embodiment.

[0027] Figure 6B Figure 6B It is a diagram showing an example of the height distribution of the array substrate detected by the scanning device in the sub-scanning direction according to the first embodiment.

[0028] Figure 7 Figure 7 It is a flowchart showing the operation of obtaining height information according to the first embodiment.

[0029] Figure 8A Figure 8A It is a diagram showing the scanning area on the array board during the operation of obtaining height information.

[0030] Figure 8B Figure 8B It is a diagram showing the positional relationship of the height scanning during the operation of obtaining height information.

[0031] Figure 8C Figure 8C It is a diagram showing the positional relationship of the height scanning during the operation of obtaining height information.

[0032] Figure 9 Figure 9 It is a diagram showing the positional relationship of the sub-scanning of the scanning device according to the second embodiment.

[0033] Figure 10 ​​​​​​​​​​​​​​​​​​​​​​​​​Figure 10 is a flowchart illustrating the operation of the scanning device according to the second embodiment.

[0034] Figure 11 Figure 11 is a diagram illustrating the internal configuration of the controller according to the third embodiment.

[0035] Figure 12 Figure 12 is a diagram illustrating the operation of the piston crank mechanism.

[0036] Figure 13 Figure 13 is a flowchart illustrating the operation of the coordinate calculation circuit according to the third embodiment. Detailed Description of the Invention

[0037] Preferred embodiments of the present invention will be described below with reference to the accompanying drawings.

[0038] <First Embodiment>

[0039] Figure 1 is a schematic diagram illustrating the configuration of the sample measurement device 100 according to the first embodiment.

[0040] The sample measurement device 100 serves as a scanning device for a scanning target. The sample measurement device 100 measures a sample that is a target located on one surface of the array plate 101. A large number of biomolecules are fixed at spots on the glass slide of the array plate 101 and are fluorescently labeled.

[0041] The light source 102 is a semiconductor laser configured to emit light with a wavelength close to 670 nm.

[0042] 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 front surface (upper surface) of the array plate 101 to the photosensor 105. The confocal optical system 103 consists of a pinhole, a filter, a dichroic mirror, a quarter-wave plate, a polarization beam splitter, and a lens. Using the confocal optical system 103 reduces the influence of the fluorescence component from the glass slide of the array plate 101 itself, thereby improving the signal-to-noise ratio in the measurement of the fluorescence component originating from the spots.

[0043] ​​​​​​The light guiding unit 104 irradiates the excitation light onto the spots on the array plate 101. The light guiding unit 104 is composed of a prism for guiding the excitation light toward the array plate 101 and a lens for focusing the excitation light onto the spots on the array plate 101. In this embodiment, the light guiding unit 104 is positioned below the array plate 101 and irradiates the excitation light upward. In addition, the light guiding unit 104 is configured to irradiate the excitation light as primary light onto the target and capture the fluorescence as secondary light. The light guiding unit 104 corresponds to an example of the observation optical system.

[0044] The light sensor 105 converts light into an electrical signal. The light sensor 105 can use a photomultiplier tube and / or a photodiode. The light sensor 105 can separately acquire the fluorescence from the spots on the array plate 101 and the reflected light from the front surface of the array plate 101. The light sensor 105 corresponds to an example of the detection unit configured to acquire optical information.

[0045] The piston crank mechanism 106 moves the light guiding unit 104 along a plane perpendicular to the optical axis of the lens of the light guiding unit 104 or the optical axis of the excitation light. Specifically, the piston crank mechanism 106 reciprocates the light guiding unit 104 along the short side direction of the array plate 101. As the light guiding unit 104 reciprocates, the excitation light from the light source 102 is scanned in the short side direction of the array plate 101. It should be noted that the short side direction of the array plate 101 is referred to as the main scanning direction, and the reciprocating scan performed by the piston crank mechanism 106 is referred to as the main scan. In other words, the main scanning direction is the direction parallel to the short side of the array plate 101. In this embodiment, the stroke of the main scan is approximately 30 mm. It should be noted that the operating direction of the light guiding unit 104 is constrained to the main scanning direction by a guide (not shown).

[0046] The pulse motor 107 rotates the piston crank mechanism 106 at a high speed. In this embodiment, the rotation speed of the pulse motor 107 is about 1200 rpm. The pulse motor 107 corresponds to an example of the driving unit.

[0047] The encoder 108 measures the position of the light guiding unit 104 in the main scanning direction. The encoder 108 is mounted on the piston crank mechanism 106 and outputs a phase difference pulse voltage composed of phases A, B, and Z based on the position of the light guiding unit 104 in the main scanning direction. The encoder 108 corresponds to an example of the measuring unit.

[0048] The linear stage 109 moves the array plate 101 along a plane perpendicular to the optical axis of the lens of the light guiding unit 104 or the optical axis of the excitation light. Specifically, the linear stage 109 moves the array plate 101 in a horizontal plane along a direction perpendicular to the main scan. The linear stage 109 consists of a ball screw and a home sensor. It should be noted that scanning in a direction perpendicular to the main scan in a horizontal plane is referred to as sub-scan. In other words, the sub-scan direction is the direction parallel to the long side of the array plate 101. There is a placement portion for placing the array plate 101 on the linear stage 109. The user pre-places the array plate 101 on the placement portion.

[0049] The pulse motor 110 is connected to the linear stage 109. The rotational motion of the pulse motor 110 is converted into a linear motion by the ball screw of the linear stage 109.

[0050] It should be noted that the piston crank mechanism 106, the pulse motor 107, the linear stage 109, and the pulse motor 110 correspond to examples of the scanning unit.

[0051] The motor driver 111 is a driver circuit for rotating the pulse motor 110. In this embodiment, inputting a pulse signal to the motor driver 111 causes the pulse motor 110 to rotate 0.72° and moves the array plate 101 2 μm in the sub-scan direction.

[0052] The linear stage 112 moves the array plate 101 in the vertical direction along the optical axis direction of the lens of the light guiding unit 104 or the optical axis direction of the excitation light. The linear stage 112 consists of a ball screw and a home sensor. It should be noted that scanning in the vertical direction is referred to as height scanning.

[0053] The pulse motor 113 is connected to the linear stage 112. The rotational motion of the pulse motor 113 is converted into a linear motion by the ball screw of the linear stage 112.

[0054] The linear stage 112 and the pulse motor 113 correspond to examples of the adjustment unit.

[0055] The motor driver 114 is a driver circuit for rotating the pulse motor 113. In this embodiment, inputting a pulse signal to the motor driver 114 causes the pulse motor 113 to rotate 0.72° and moves the array plate 101 1 μm upward (i.e., in the vertical direction).

[0056] The motor driver 115 is a driver circuit for rotating the pulse motor 107. In this embodiment, inputting a pulse signal to the motor driver 115 causes the pulse motor 107 to rotate 0.72° and moves the light guiding unit 104 along the main scan direction.

[0057] The controller 116 controls the entire sample measurement device 100. The controller 116 consists of a field programmable gate array (FPGA), a central processing unit (CPU), and a memory, and functions as a computer. The controller 116 performs main scanning, sub-scanning, and height scanning of the excitation light on the array board 101 by controlling the light source 102, the motor driver 111, the motor driver 114, and the motor driver 115. In addition, during the scanning, the controller 116 acquires fluorescence signal data (two-dimensional image) based on the position information of the light guiding unit 104 measured by the encoder 108 and the output signal from the light sensor 105, and stores the acquired fluorescence signal data (two-dimensional image) in the internal memory.

[0058] As described below, the controller 116 acquires height information and tilt information about the array board 101, and generates drive pulse sequences output to the motor drivers 111 and 114 during sub-scanning and height scanning. In addition, the controller 116 controls the timing of sub-scanning and height scanning and the timing of fluorescence signal data acquisition in synchronization with the position information of the light guiding unit 104. By performing the above control, when the light guiding unit 104 is outside the imaging area on the array board 101, the thickness and tilt of the array board 101 are corrected so that height scanning can be performed to focus the entire surface of the array board 101.

[0059] The user interface 117 is an interface for receiving instructions from the user and displaying results. The user interface 117 consists of a keyboard, a mouse, and a display.

[0060] The controller 116 can receive an imaging instruction from the user via the user interface 117, and present image data based on the fluorescence signal data to the user via the user interface 117. In addition, the user can specify the imaging area, the pixel pitch in the main scanning direction, and the pixel pitch in the sub-scanning direction via the graphical user interface (GUI) on the user interface 117 during imaging.

[0061] The piston crank mechanism 106 includes a crank 118 and a connecting rod 119.

[0062] The crank 118 is connected to the rotating shaft of the pulse motor 107 and the connecting rod 119 by joints. The crank 118 has a length r.

[0063] The connecting rod 119 is connected to the crank 118 and the light guiding unit 104 by joints. The connecting rod 119 has a length l.

[0064] Figure 2A is a diagram showing the array board 101 viewed from above, and Figure 2B is a diagram showing the array board 101 viewed from the side.

[0065] The array plate 101 is composed of a glass slide 201 having a rectangular shape with a short side and a long side, and a plurality of spots 202 arranged on the upper surface. At each spot 202, a biomolecule containing a peptide bond is fixed. Here, a single type of biomolecule is fixed at a single spot 202.

[0066] In this embodiment, the diameter of each spot 202 is about 100 um, and the spot pitch is 200 um. In addition, the length of the array plate 101 in the short side direction (short side) is 25 mm, and the length in the long side direction (long side) is 75 mm. The point 203 at the upper left of the array plate 101 is defined as the origin, the rightward direction in the short side direction is defined as the positive direction of the X axis, and the downward direction in the long side direction is defined as the positive direction of the Y axis. Assuming that the X coordinate and the Y coordinate are in um units, the coordinates of the four corners of the array plate 101 are (0, 0), (25000, 0), (0, 75000), and (25000, 75000).

[0067] 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. In other words, in the main scan, an additional 2.5 mm is scanned on both the left and right sides of the array plate 101, and the X coordinate of the scanning range extends from -2500 to 27500.

[0068] Due to limitations in spot generation or considering user grasping, the glass slide 201 includes a region where the spots 202 exist and a region where the spots 202 do not exist. Region 204 is the region on the glass slide 201 where the spots 202 exist. In this embodiment, the coordinates of the four corners of region 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.

[0069] Figure 3 It is a block diagram showing the internal configuration of the controller 116 according to the first embodiment.

[0070] The CPU 301 executes software (program) configured to control the entire controller 116. The CPU 301 is composed of a microprocessor and a cache memory. The CPU 301 corresponds to an example of a control unit.

[0071] The bus interface 302 is an interface for connecting the CPU 301 to various peripheral circuits.

[0072] The memory 303 stores the imaging conditions input by the user, the parameters of the sample measurement device 100, and the fluorescence signal data. The memory 303 can use a double data rate 4 synchronous dynamic random access memory (DDR4-SDRAM) and / or a solid state drive (SSD). The memory corresponds to an example of a storage unit.

[0073] The memory control circuit 304 controls the memory 303 based on commands to access the memory 303 via the bus interface 302.

[0074] 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 and a digital-to-analog (DA) converter. The CPU 301 can control the on / off state and light intensity of the laser irradiation from the light source 102 via the light source control circuit 305.

[0075] The data acquisition circuit 306 is a circuit configured to acquire fluorescence signal data based on instructions from the CPU 301, based on the output signal from the optical sensor 105 and the position of the light guiding unit 104 relative to the array board 101, and continuously store the acquired fluorescence signal data in the memory 303. The data acquisition circuit 306 consists of a buffer circuit, an analog-to-digital (AD) converter, an AD converter control circuit, and a direct memory access (DMA) controller. The data acquisition circuit 306 corresponds to an example of an image acquisition unit.

[0076] The motor control circuit 307 generates a control signal for the motor driver 115 of the pulse motor 107 used as the main scanning motor based on instructions from the CPU 301. The motor control circuit 307 generates a drive pulse voltage based on instructions from the CPU 301 regarding the rotational speed, acceleration, displacement, rotational direction, and rotation start timing of the pulse motor 107.

[0077] The motor control circuit 308 generates a control signal for the motor driver 111 of the pulse motor 110 used as the sub-scanning motor based on instructions from the CPU 301. The motor control circuit 308 generates a drive pulse voltage based on instructions from the CPU 301 regarding the rotational speed, acceleration, displacement, rotational direction, and rotation start timing of the pulse motor 110.

[0078] The motor control circuit 309 generates a control signal for the motor driver 114 of the pulse motor 113 used as the height scanning motor based on instructions from the CPU 301. The motor control circuit 309 generates a drive pulse voltage based on instructions from the CPU 301 regarding the rotational speed, acceleration, displacement, rotational direction, and rotation start timing of the pulse motor 113.

[0079] The coordinate calculation circuit 310 counts the phase difference pulse signals of two phases A and B from the encoder 108, and calculates the position of the light guiding unit 104. In this embodiment, the encoder 108 has a resolution of 1um. When the level of the phase A signal or the phase B signal changes and the phase A signal is ahead of the phase B signal in phase, the coordinate calculation circuit 310 increases the coordinate of the light guiding unit 104 by 1um. In addition, when the level of the phase A signal or the phase B signal changes and the phase B signal is ahead of the phase A signal in phase, the coordinate calculation circuit 310 decreases the coordinate of the light guiding unit 104 by 1um.

[0080] The synchronization circuit 311 generates trigger signals for the data acquisition circuit 306, the motor control circuit 308, and the motor control circuit 309 based on the coordinate information calculated by the coordinate calculation circuit 310. Here, the trigger signal for the data acquisition circuit 306 is called the data acquisition trigger signal, the trigger signal for the motor control circuit 308 is called the sub-scanning trigger signal, and the trigger signal for the motor control circuit 309 is called the height scanning trigger signal.

[0081] After receiving the data acquisition trigger signal, the data acquisition circuit 306 controls the internal AD converter, acquires a single piece of data output by the optical sensor 105, and stores the acquired output data in the memory 303 via the internal DMA controller.

[0082] After receiving the sub-scanning trigger signal, the motor control circuit 308 outputs a drive pulse sequence corresponding to the displacement of the pixel pitch in the sub-scanning direction to the motor driver 111. The pixel pitch in the sub-scanning direction is specified by the user at the start of imaging and stored in the memory 303.

[0083] After receiving the height scanning trigger signal, the motor control circuit 309 outputs a drive pulse sequence corresponding to the displacement in the height scanning direction to the motor driver 114. The displacement in the height scanning direction is calculated by the CPU 301. The calculation method will be described below.

[0084] The communication circuit 312 is a circuit for connecting the sample measurement device 100 to an external network. A communication method using a communication protocol conforming to the Ethernet standard is adopted. Connecting the sample measurement device 100 to an external personal computer (external PC) or an external server enables remote control of imaging and data storage in an external mass storage.

[0085] The user interface (UI) circuit 313 is a circuit for connecting the sample measurement device 100 to the user interface 117. The UI circuit 313 consists of an input circuit from a keyboard and a mouse and an image forming circuit for controlling a display.

[0086] It should be noted that the peripheral circuit constituting the controller 116 is implemented on a semiconductor chip such as an FPGA or an ASIC, and operates in synchronization with a clock. In this embodiment, the clock frequency is 100 MHz.

[0087] Figure 4 is a flowchart illustrating operations in the imaging process performed by the sample measurement device 100 according to the first embodiment. Figure 4 The flowchart in is implemented by the CPU 301 of the controller 116 executing a program.

[0088] Figures 5A to 5C is a diagram illustrating the positional relationship of sub-scanning on the array board 101 in the imaging process and the array board 101 as viewed from above.

[0089] Figure 6A and Figure 6B is a diagram illustrating the positional relationship of height scanning on the array board 101 in the imaging process and the array board 101 as viewed from the side (long side). The Z = 0 position on the vertical axis represents the horizontal reference plane. Figure 6A and Figure 6B differ in terms of the thickness and tilt of the placed array board.

[0090] In S401, the CPU 301 reads imaging conditions based on an imaging instruction from the user. The user inputs imaging conditions in advance via the user interface 117. The CPU 301 stores the acquired imaging conditions in the memory 303. Here, the points 501(X1, Y1) and 502(X2, Y2) indicating the imaging area on the array board 101, the pixel pitch Xp in the main scanning direction, the pixel pitch Yp in the sub-scanning direction, and the rotational speed Xs in the main scanning direction are input as imaging conditions. In this embodiment, X1 = 500, X2 = 22500, Y1 = 500, Y2 = 64500, Xp = 10um, Yp = 10um, and Xs = 1200 rpm. In other words, the memory 303 is a storage unit configured to store information regarding the imaging area defined with respect to the surface (one surface) of the array board 101 including a plurality of spots 202.

[0091] The CPU 301 sets the imaging conditions in the synchronization circuit 311. Here, the rectangular area with points 501 and 502 as the diagonals on the array board 101 is referred to as the imaging area 503. In addition, 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 this embodiment, Nx = 2200, and Ny = 6400.

[0092] In S402, the CPU 301 obtains height information regarding the array board 101 and obtains 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 board. The height from the horizontal reference plane to the front surface of the array board 101 (i.e., the surface having the dots 202) is referred to as height information. Further, the tilt of the array board 101 with respect to the horizontal reference plane in the sub-scanning direction is referred to as tilt information. Here, the CPU 301 obtains the height information Z3 and Z4 by measuring two Y coordinates Y3 and Y4. In the present embodiment, Y3 = 750, and Y4 = 65000.

[0093] In the case of the tilt as Figure 6A shown, Z4 > Z3, while in the case of the tilt as Figure 6B shown, Z3 < Z4. The method for obtaining height information will be described below with reference to Figure 7 the flowchart shown in

[0094] The tilt information K is calculated using the following (Equation 1):

[0095] K = (Z4 - Z3) / (Y4 - Y3) (Equation 1).

[0096] In S403, the CPU 301 calculates the target height of the sub-scanning position (for each line) from the reference plane based on the imaging conditions, the height information, and the tilt information K. Here, the target height Z(Y) of the sub-scanning position at the coordinate Y is given by the following (Equation 2):

[0097] Z(Y) = K × (Y - Y3) + Z3 (Equation 2).

[0098] In Figure 6A and Figure 6B the target height 601 corresponds to the Z coordinate of the front surface of the array board 101.

[0099] In S404, the CPU 301 instructs the motor control circuits 307, 308, and 309 to move the array board 101 and the light guiding unit 104 to the imaging start position.

[0100] In the present embodiment, the X coordinate of the imaging start position is at the end of the scanning range of the piston crank mechanism 106, and the X coordinate value is -2500. The Y coordinate of the imaging start position is Y1 specified by the imaging conditions. Further, the Z coordinate Z1 of the imaging start position is calculated using Z(Y1) in (Equation 1).

[0101] In S405, the CPU 301 starts the main scanning (scanning process) of the light guiding unit 104 in the main scanning direction. Specifically, the CPU 301 instructs the motor control circuit 307 to rotate the pulse motor 107 at a rotation speed Xs. When the pulse motor 107 rotates, the light guiding unit 104 starts reciprocating movement in the X direction. The X coordinate of the light guiding unit 104 is calculated by the encoder 108 and the coordinate calculation circuit 310 for each clock and is output to the synchronization circuit 311.

[0102] In S406, the CPU 301 instructs the light source control circuit 305 to cause the light source 102 to start emitting light. When the light source 102 emits light, the irradiation of the array plate 101 starts through the light guiding unit 104.

[0103] In S407, the synchronization circuit 311 determines whether the light guiding unit 104 has reached the line feed position. In the case where the X coordinate of the light guiding unit 104 output from the coordinate calculation circuit 310 moves from the imaging area to outside the imaging area, the synchronization circuit 311 determines that the light guiding unit 104 has reached the line feed position. In the case where the current main scanning direction is the forward direction (the direction in which the X coordinate increases), when the X coordinate of the light guiding unit 104 exceeds X2, it is determined that the light guiding unit 104 has reached the line feed position. The scanning in the forward direction is represented by Figure 5B the trajectory 504 in. On the other hand, in the case where the current main scanning direction is the return direction (the direction in which the X coordinate decreases), when the X coordinate of the light guiding unit 104 becomes less than X1, the synchronization circuit 311 determines that the light guiding unit 104 has reached the line feed position. The scanning in the return direction is represented by Figure 5B the trajectory 506 in. The initial value of the main scanning direction is the forward direction, and thereafter, the return direction and the forward direction alternate every time the line feed position is reached.

[0104] In the case where the line feed position has been reached, the synchronization circuit 311 outputs a sub-scanning trigger signal and a height scanning trigger signal to execute S416 in parallel with S417 and S418. In the case where the line feed position has not been reached, the synchronization circuit 311 does not output a sub-scanning trigger signal or a height scanning trigger signal, and the process proceeds to S408.

[0105] In S408, the synchronization circuit 311 determines whether the light guiding unit 104 has reached the sampling position. The sampling position refers to the point on the array plate 101 where the fluorescence signal data is acquired. It should be noted that although the case where the sampling position is different from the spot is described, the sampling position can be the same as the spot. The X coordinate P(N) of the Nth sampling position is represented by the following (Equation 3):

[0106] P(N) = X1 + Xp × (N + 1 / 2) (N = 0, 1,..., Nx - 1) (Equation 3).

[0107] The sampling positions refer to multiple points on the trajectory, such as Figure 5B point 508 in, and the pitch in the X direction and the Y direction are Xp and Yp, respectively. Further, the initial value of the sampling position is P(0), and it is stored in the synchronization circuit 311. In the first sampling position determination, when the X coordinate of the light guiding unit 104 output from the coordinate calculation circuit 310 has passed P(0) in the forward direction (the direction in which the X coordinate increases), it is determined that the light guiding unit 104 has reached the sampling position. In the second and subsequent sampling position determinations, when the X coordinate of the light guiding unit 104 output from the coordinate calculation circuit 310 has passed the sampling position updated in S410 below, it is determined that the light guiding unit 104 has reached the sampling position.

[0108] When the sampling position has been reached, the synchronization circuit 311 outputs a data acquisition trigger signal, and the process proceeds to S409. When the sampling position has not been reached, the synchronization circuit 311 does not output a data acquisition trigger signal, and the process proceeds to S411.

[0109] In S409, the data acquisition circuit 306 acquires optical information from the sampling position. Specifically, the data acquisition circuit 306 outputs a conversion start signal of the internal AD converter and performs AD conversion of the output voltage from the optical sensor 105. The AD-converted fluorescence signal data is stored in the memory 303 via the internal DMA controller of the data acquisition circuit 306 and the memory control circuit 304. After Nx×Ny pieces of data are stored in the memory 303, the data acquisition circuit 306 sets the internal data acquisition completion register to "1", and when Nx×Ny pieces of data are not stored in the memory 303, the data acquisition circuit 306 sets the internal data acquisition completion register to "0". The process of S409 is executed while the light guiding unit 104 reciprocates in the main scanning direction, but the process of S409 is not executed while the light guiding unit 104 reciprocates in the sub-scanning direction.

[0110] In S410, the synchronization circuit 311 updates the sampling position stored internally. When the current main scanning direction is the forward direction (the direction in which the X coordinate increases), the synchronization circuit 311 updates the sampling position P(N) to P(N + 1), and when the current main scanning direction is the return direction (the direction in which the X coordinate decreases), the synchronization circuit 311 updates the sampling position P(N) to P(N - 1).

[0111] In S411, the CPU 301 determines whether data acquisition has been completed. The CPU 301 reads the data acquisition completion register of the data acquisition circuit 306, and when the value of the data acquisition completion register is 1, the CPU 301 determines that data acquisition has been completed, and the process proceeds to S412. When the value of the data acquisition completion register is 0, the CPU 301 determines that data acquisition has not been completed, and the process proceeds to S407.

[0112] In S412, the CPU 301 instructs the light source control circuit 305 to stop the emission of light from the light source 102. When the light source 102 stops emitting light, the irradiation of the array board 101 by the light guide unit 104 stops.

[0113] 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 movement of the light guide unit 104 in the X direction stops.

[0114] In S414, the CPU 301 instructs the motor control circuits 307, 308, and 309 to move the array board 101 and the light guide unit 104 to the stop position. The X coordinate, Y coordinate, and Z coordinate of the stop position are 0. The movement to the stop position is performed by returning each axis to the origin using the Z-phase pulse signal of the encoder 108, the origin sensor signal in the linear stage 109, and the origin sensor signal of the linear stage 112.

[0115] In S415, the CPU 301 reads the Nx×Ny pieces of fluorescence signal data stored in the memory 303, performs data compression and format conversion processing, and generates a fluorescence image file in the Tagged Image File Format (TIFF). The fluorescence image file is stored in the memory 303 and presented to the user via the UI circuit 313 and the user interface 117. In addition, based on an instruction from the user, it is sent to an external data server via the communication circuit 312.

[0116] In S416, the CPU 301 performs a sub-scan (scanning process) of moving the array board 101 in the sub-scan direction. Specifically, the CPU 301 instructs the motor control circuit 308 to move the array board 101 by Yp in the sub-scan direction. Here, the number of pulses output from the motor control circuit 308 to the motor driver 111 is Yp / My, where My is the displacement of the array board 101 when one pulse of the voltage pulse signal is sent to the motor driver 111. In this embodiment, My = 2um. The sub-scan is represented by Figure 5B the trajectories 505 and 507 in, and the moving distance in the Y direction is Yp.

[0117] Figure 5C It is an enlarged view of trajectories 505 and 507. Trajectory 505 includes a linear trajectory 511 in the forward direction along the main scanning direction, a semi-circular trajectory 512, and a linear trajectory 513 in the return direction along the main scanning direction. In addition, trajectory 507 includes a linear trajectory 514 in the return direction along the main scanning direction, a semi-circular trajectory 515, and a linear trajectory 513 in the forward direction along the main scanning direction. As described above, each of the trajectories 505 and 507 in this embodiment includes trajectories corresponding to at least two moving directions.

[0118] Based on an instruction from the CPU 301, the motor control circuit 308 outputs a pulse signal to the motor driver 111 at a speed that allows the completion of sub-scanning when the light guiding unit 104 is outside the imaging area in the main scanning direction.

[0119] In S417, the CPU 301 reads the target heights before and after sub-scanning from the memory 303 to perform height scanning. Specifically, the CPU 301 reads Z(Y + Yp) and Z(Y), and calculates the displacement in height scanning. The displacement in height scanning varies depending on the current Y coordinate, but the number of output pulses and the rotation direction of the pulse motor 113 are calculated such that the height in the Z direction after movement is closest to the target height Z(Y + Yp) at the Y coordinate after sub-scanning in S416.

[0120] The specific calculation direction will be described below.

[0121] Here, the displacement of the array board 101 when one pulse of the voltage pulse signal is sent to the motor driver 114 is represented as Mz. In this embodiment, Mz = 1um.

[0122] RoundMz(x) represents the multiple of Mz closest to the number x, ABS(x) represents the absolute value of the number, and Sign(x) represents the sign of the 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 Figure 6A and Figure 6B the height 602 in. The number of pulses output by the motor control circuit 309 to the motor driver 114 is represented by the following (Equation 4):

[0123] Zp = ABS(RoundMz(Z(Y + Yp)) - RoundMz(Z(Y))) (Equation 4).

[0124] RoundMz(Z(Y)) takes discrete values close to the front surface of the array board 101 as specified by the height 602 in Figure 6A and Figure 6B the height 602.

[0125] In addition, the moving direction Dir of the array plate in the height direction is represented by the following (Equation 5):

[0126] Dir = Sign(RoundMz(Z(Y + Yp)) - RoundMz(Z(Y))) (Equation 5).

[0127] The positive direction of Dir is defined as vertically upward, in the direction in which the distance between the light guide unit 104 and the array plate 101 increases. In the case where the inclination of the array plate 101 is as Figure 6A shown, the value of Dir is 1, while in the case where the inclination of the array plate 101 is as Figure 6B shown, the value of Dir is -1.

[0128] In S418, the CPU 301 performs a height scan to adjust the array plate 101 in the vertical direction in order to acquire focusing optical information (adjustment process). Specifically, the CPU 301 instructs the motor control circuit 309 to move the array plate 101 by Zp in the moving direction Dir. When the value of Dir is positive, the array plate 101 moves upward, while when the value of Dir is negative, the array plate 101 moves downward. The process of S418 is executed based on the scan sequence information indicating that the light guide unit 104 has reached the line feed position in S407. In other words, the CPU 301 determines whether to perform a height scan based on the scan sequence information.

[0129] The number of pulses output from the motor control circuit 309 to the motor driver 111 is Zp / Mz. Based on an instruction from the CPU 301, the motor control circuit 309 outputs a pulse signal to the motor driver 114 at a speed that allows the completion of the height scan when the light guide unit 104 is outside the imaging area in the main scanning direction. Therefore, the height scan is performed during the sub-scanning period. In other words, the height scan and the sub-scan are performed in parallel. On the other hand, the height scan is not performed during the main scanning period.

[0130] When the sub-scan in S416 and the height scan in S418 are completed, the process proceeds to S419.

[0131] In S419, the synchronization circuit 311 updates the current main scanning direction and the Y coordinate. In other words, when the previous main scanning direction was the forward direction, the synchronization circuit 311 updates the main scanning direction to the return direction and updates the line feed position to X1. On the other hand, when the previous main scanning direction was the return direction, the synchronization circuit 311 updates the main scanning direction to the forward direction and updates the line feed position to X2. In addition, the synchronization circuit 311 increments the previous Y coordinate by Yp to update the current Y coordinate, and the process proceeds to S411.

[0132] Figure 7 is a flowchart showing the operation of obtaining height information about the array plate 101, which corresponds to a part of the process of S402 described above. It should be noted that, in Figure 7 the process shown in the flowchart of Figure 4 , different from the process shown in the flowchart of

[0133] Figure 8A and Figure 8B are diagrams showing the positional relationship of height scanning on the array plate 101 in the operation of obtaining height information and the array plate 101 as viewed from the side (short side). Figure 8C The intensity of the reflected light acquired by the light sensor 105 during height scanning is plotted for each height, where the horizontal axis represents the magnitude of the light intensity and the vertical axis represents the acquired height.

[0134] In S701, the CPU 301 sets parameters for height information acquisition in the synchronization circuit 311. The Y coordinate Yh of the position where height information acquisition is performed, the pixel pitch Xp in the main scanning direction, the pixel pitch Zp in the height scanning direction, the points 801 (X5, Z5) and 802 (X6, Z6) indicating the height scanning range in the XZ plane, and the rotational speed Xs in the main scanning direction are set as parameters.

[0135] The rectangular area 803 with points 801 and 802 as its diagonals in the XZ plane is called the height scanning area. The CPU 301 pre - calculates the number of pixels Nx = (X6 - X5) / Xp in the main scanning direction and the number of pixels Nz = (Z6 - Z5) / Zp in the height scanning direction. 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.

[0136] In S702, the CPU 301 instructs the motor control circuits 307, 308, and 309 to move the array plate 101 and the light guiding unit 104 to the height information acquisition start position.

[0137] In this embodiment, the X coordinate of the height information acquisition start position is located at the end of the scanning range of the piston - crank mechanism 106, and the X coordinate value is - 2500. As specified by the parameters, the Y coordinate of the height information acquisition start position is Yh. In addition, as specified by the parameters, the Z coordinate of the height information acquisition start position is Z5.

[0138] In S703, the CPU 301 starts the main scan. This process is similar to the process of S405 described above.

[0139] In S704, the CPU 301 causes the light source 102 to start light emission, thereby starting light irradiation. This process is similar to the process of S406 described above.

[0140] In S705, the synchronization circuit 311 determines whether the light guide unit 104 has reached the line feed position. When the X coordinate of the light guide unit 104 output from the coordinate calculation circuit 310 moves out of the imaging area from the imaging area, the synchronization circuit 311 determines that the light guide unit 104 has reached the line feed position. When the current main scan direction is the forward direction (the direction in which the X coordinate increases), when the X coordinate of the light guide unit 104 exceeds X6, it is determined that the light guide unit 104 has reached the line feed position. The scan in the forward direction is represented by Figure 8B the trajectory 804 in. On the other hand, when the current main scan direction is the return direction (the direction in which the X coordinate decreases), when the X coordinate of the light guide unit 104 becomes less than X5, the synchronization circuit 311 determines that the light guide unit 104 has reached the line feed position. The scan in the return direction is represented by Figure 8B the trajectory 806 in. The initial value of the main scan direction is the forward direction, and thereafter, each time the line feed position is reached, the return direction and the forward direction alternate.

[0141] When the line feed position has been reached, the synchronization circuit 311 outputs a height scan trigger signal, and the process proceeds to S714. When the line feed position has not been reached, the synchronization circuit 311 does not output a height scan trigger signal, and the process proceeds to S706.

[0142] In S706, the synchronization circuit 311 determines whether the light guide unit 104 has reached the sampling position. This process is similar to the process of S408 described above. The sampling position refers to multiple points on the trajectory, such as Figure 8B the point 808 in, and the pitch in the X direction and the Z direction are Xp and Zp, respectively. When the sampling position has been reached, the synchronization circuit 311 outputs a data acquisition trigger signal, and the process proceeds to S707. When the sampling position has not been reached, the synchronization circuit 311 does not output a data acquisition trigger signal, and the process proceeds to S709.

[0143] In S707, the data acquisition circuit 306 acquires reflected light signal data from the sampling position. This process is similar to the process of S409 described above. After Nx × Nz pieces of data are stored in the memory 303, the data acquisition circuit 306 sets the internal data acquisition completion register to "1", and in the case where Nx × Nz pieces of data are not stored in the memory 303, the data acquisition circuit 306 sets the internal data acquisition completion register to "0".

[0144] In S708, the synchronization circuit 311 updates the internally stored sampling position. This process is similar to the process of S410 described above.

[0145] In S709, the CPU 301 determines whether data acquisition has been completed. This process is similar to the process of S411 described above. When the CPU 301 determines that data acquisition has been completed, the process proceeds to S710. When the CPU 301 determines that data acquisition has not been completed, the process proceeds to S705.

[0146] In S710, the CPU 301 stops the emission of light from the light source 102. This process is similar to the process of S412 described above.

[0147] In S711, the CPU 301 stops the main scan. This process is similar to the process of S413 described above.

[0148] In S712, the CPU 301 moves the array board 101 and the light guiding unit 104 to the stop position. This process is similar to the process of S414 described above.

[0149] In S713, the CPU 301 reads Nx × Nz pieces of reflected light signal data stored in the memory 303, analyzes the read data, and calculates height information about the array board. Specifically, Nx pieces of data acquired at the same height are summed and then averaged to calculate the average light intensity at each height. When the average light intensity is arranged according to the Z coordinate, there are two peaks corresponding to the front surface and the back surface of the array board 101. In Figure 8C the peak 809 indicates the peak caused by the reflected light from the front surface of the array board 101, and the peak 810 indicates the peak caused by the reflected light from the back surface of the array board 101. The Z coordinate 811 indicating the peak 809 with the larger Z coordinate among the two peaks (i.e., the peak corresponding to the front surface) corresponds to the height information at the position Yh.

[0150] In S714, the CPU 301 instructs the motor control circuit 309 to move the array board 101 by Zp in the height scanning direction. The number of pulses output from the motor control circuit 309 to the motor driver 114 is Zp / Mz, where Mz is the displacement of the array board 101 when one pulse of the voltage pulse signal is sent to the motor driver 114. The height scanning is represented by the trajectories 805 and 807 in Figure 8B and the moving distance in the Z direction is Zp.

[0151] Based on the instruction from the CPU 301, when the light guiding unit 104 is outside the imaging area in the main scanning direction, the motor control circuit 309 outputs a pulse signal to the motor driver 114 at a speed that allows the height scanning to be completed.

[0152] In S715, the synchronization circuit 311 updates the current main scanning direction and Z coordinate. In other words, when the previous main scanning direction is the forward direction, the synchronization circuit 311 updates the main scanning direction to the return direction and updates the line feed position to X5. On the other hand, when the previous main scanning direction is the return direction, the synchronization circuit 311 updates the main scanning direction to the forward direction and updates the line feed position to X6. In addition, the synchronization circuit 311 increments the previous Z coordinate by Zp to update the current Z coordinate, and the process proceeds to S709.

[0153] When obtaining the tilt information in S402 described above, the processing in the flowchart in Figure 7 is performed for each of the Y coordinates Y3 and Y4 to obtain the height information Z3 for the Y coordinate Y3 and the height information Z4 for the Y coordinate Y4, and the tilt information K is calculated using (Equation 1). By calculating the tilt information K as described above, the target height can be calculated for each sub-scanning position in S403.

[0154] As described above, the sample measurement device 100 according to the present embodiment performs height scanning during the sub-scanning period to adjust the array board 101 in the vertical direction. Therefore, the focusing optical information can be obtained in a short time.

[0155] In addition, in the present embodiment, the synchronization circuit 311 is used to perform sub-scanning and height scanning in synchronization with the position of the light guiding unit 104. Since the sub-scanning and height scanning are performed when the light guiding unit 104 is outside the imaging area, it is not necessary to move the array board 101 within the imaging area. Therefore, the influence of vibration caused by driving the array board 101 can be reduced. In addition, since the sample measurement device 100 does not include a high-speed servo control system composed of a high-performance focus sensor and a low-vibration actuator, a focused fluorescence image across the entire surface of the array board 101 can be obtained with a simple configuration.

[0156] In addition, in the present embodiment, the pulse motor 107 for main scanning rotates at a constant speed during imaging to move the light guiding unit 104, and performs sub-scanning and height scanning when the light guiding unit 104 is outside the imaging region. Since it is not necessary to temporarily stop the pulse motor 107 for main scanning before sub-scanning and height scanning, reciprocating scanning of the light guiding unit 104 can be performed at high speed, which reduces the imaging time.

[0157] In addition, in the present embodiment, two-dimensional scanning involving both main scanning and sub-scanning is performed while correcting individual differences in the thickness and tilt of the entire array plate 101 based on at least height information and tilt information acquired in advance at two points. Therefore, compared with three-dimensional scanning of the array plate 101, a focused fluorescence image across the entire surface of the array plate 101 can be acquired in a short time.

[0158] In addition, in the present embodiment, for each sub-scanning position (each row), the number of target pulses closest to the target height is calculated. Therefore, in the case where the amount and direction of height scanning are not predetermined values, especially when the displacement of each row is different, the height can be adjusted to a height close to the target height. Therefore, even when it is difficult to predict in advance the height and tilt of the array plate 101 due to individual differences in the thickness and tilt of the array plate 101 and the method of placing the array plate 101, a focused fluorescence image across the entire surface of the array plate can be obtained.

[0159] In addition, in the present embodiment, main scanning is also performed during height information acquisition, and Nx pieces of data are averaged, so that stable peak detection can be achieved even when there is local contamination or liquid on the array plate 101. Therefore, compared with detecting peaks at a single point on the array plate 101 for each height scanning position, the accuracy of the acquired height information can be improved.

[0160] In addition, in the present embodiment, the synchronization circuit 311 is used to sample fluorescence signal data synchronously with the position of the light guiding unit 104. Compared with the case where data sampling and sub-scanning are performed at constant intervals without synchronizing with the position of the light guiding unit 104, data at uniform intervals can be acquired across the entire array plate 101, thereby improving the position accuracy when acquiring a captured image.

[0161] It should be noted that although the present embodiment describes the case where the imaging region 503 covers the entire region 204 and all the spots on the array plate 101 are imaged, this is not a restrictive case. For example, the user can set a part of the region 204 as the imaging region 503. In this case, by scanning only the part including the spots of interest to the user, the imaging time can be reduced.

[0162] It should be noted that although this embodiment describes the case of obtaining height information using the peak of the reflected light from the front surface of the glass slide 201, this is not a restrictive case. For example, the peak position of the fluorescence signal brightness from some spots on the array plate 101 can be used to obtain height information. In this case, although it is necessary to irradiate the spots with light to obtain height information, since the optical sensor 105 does not need to obtain the reflected light, the number of components in the optical system can be reduced.

[0163] It should be noted that although this embodiment describes the case of a single wavelength from the light source 102, multiple light sources, multiple optical systems, and multiple optical sensors for each wavelength can be included, and the array plate 101 can be irradiated with excitation light having multiple wavelengths. By comparing the fluorescence signals generated by the excitation light having multiple wavelengths, the properties of the biomolecules at the spots can be analyzed in more detail.

[0164] It should be noted that although this embodiment describes the case where the light guiding unit 104 moves in the main scanning direction, this is not a restrictive case. For example, the array plate 101 can move in the main scanning direction, or both the light guiding unit 104 and the array plate 101 can move in the main scanning direction. In other words, at least one of the light guiding unit 104 and the array plate 101 can move relative to the other in the main scanning direction.

[0165] In addition, although this embodiment describes the case where the array plate 101 moves in the sub-scanning direction, this is not a restrictive case. For example, the light guiding unit 104 can move in the sub-scanning direction, or both the light guiding unit 104 and the array plate 101 can move in the sub-scanning direction. In other words, at least one of the light guiding unit 104 and the array plate 101 can move relative to the other in the sub-scanning direction.

[0166] In addition, although this embodiment describes the case where the array plate 101 moves in the vertical direction, this is not a restrictive case. For example, the light guiding unit 104 can move in the vertical direction, or both the light guiding unit 104 and the array plate 101 can move in the vertical direction. In other words, at least one of the light guiding unit 104 and the array plate 101 can move relative to the other in the vertical direction to adjust the relative position of the light guiding unit 104 and the array plate 101.

[0167] <Second Embodiment>

[0168] The difference between the second embodiment and the first embodiment lies in the scanning method of the light guiding unit 104 and the fluorescence signal data sampling method. In the first embodiment, signal acquisition from the light sensor 105 is performed in the forward direction and the return direction of the main scan, and the sub-scan and the height scan are performed during the period outside the imaging regions at both ends of the array plate 101. In this embodiment, signal acquisition from the light sensor 105 is performed in the forward direction during the main scan, and the sub-scan and the height scan are performed in the return direction during the main scan.

[0169] It should be noted that the configuration of the sample measurement device 100 in this embodiment, as well as the internal configurations of the array plate 101 and the controller 116, are similar to those in Figure 1 , Figure 2A , Figure 2B and Figure 3 , so the descriptions of these will be omitted.

[0170] Figure 9 FIG. is a diagram illustrating the positional relationship of the sub-scan on the array plate 101 in the imaging process. Figure 10 FIG. is a flowchart of the operations in the process of imaging the array plate 101 performed by the sample measurement device 100 according to this embodiment.

[0171] S1001 to S1006 are similar to S401 to S406 in the first embodiment.

[0172] In S1007, the synchronization circuit 311 determines whether the light guiding unit 104 has reached the line feed position. When the X coordinate of the light guiding unit 104 output from the coordinate calculation circuit 310 moves from the imaging region to outside the imaging region, the synchronization circuit 311 determines that the light guiding unit 104 has reached the line feed position. When the current main scan direction is the forward direction (the direction in which the X coordinate increases), when the X coordinate of the light guiding unit 104 exceeds X2, it is determined that the light guiding unit 104 has reached the line feed position. The scan in the forward direction is represented by the trajectory 901 in Figure 9 . On the other hand, when the current main scan direction is the return direction (the direction in which the X coordinate decreases), when the X coordinate of the light guiding unit 104 becomes less than X1, the synchronization circuit 311 determines that the light guiding unit 104 has reached the line feed position. The scan in the return direction is represented by the trajectory 902 in Figure 9 . The initial value of the main scan direction is the forward direction, and thereafter, the return direction and the forward direction alternate each time the line feed position is reached.

[0173] When the line feed position has been reached, the process proceeds to S1017. When the line feed position has not been reached, the process proceeds to S1008.

[0174] In S1008, the synchronization circuit 311 determines whether the current main scanning direction is the forward direction or the return direction. If it is the forward direction, the process proceeds to S1009. If it is the return direction, the process proceeds to S1012.

[0175] In S1009, the synchronization circuit 311 determines whether the light guiding unit 104 has reached the sampling position. The sampling position refers to the points on the array board 101 where the fluorescence signal data is acquired. The X coordinate P(N) of the Nth sampling position is represented by the above (Equation 3). The sampling position refers to multiple points on the trajectory, such as Figure 9 the point 903 in, and the pitch in the X direction and the Y direction are Xp and Yp respectively, which are the same as the coordinates in the first embodiment. In addition, the initial value of the sampling position is P(0) and is stored in the synchronization circuit 311. In the first sampling position determination, when the X coordinate of the light guiding 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), it is determined that the light guiding unit 104 has reached the sampling position. In the second and subsequent sampling position determinations, when the X coordinate of the light guiding unit 104 output from the coordinate calculation circuit 310 passes through the sampling position updated in S1011 described below in the forward direction, it is determined that the light guiding unit 104 has reached the sampling position.

[0176] If the sampling position has been reached, the synchronization circuit 311 outputs a data acquisition trigger signal, and the process proceeds to S1010. If the sampling position has not been reached, the synchronization circuit 311 does not output a data acquisition trigger signal, and the process proceeds to S1012.

[0177] The processing of S1010 is similar to that of S409 in the first embodiment.

[0178] In S1011, the synchronization circuit 311 updates the internally stored sampling position. Since the current main scanning direction is the forward direction (the direction in which the X coordinate increases), the synchronization circuit 311 updates the sampling position P(N) to P(N + 1). However, when N = Nx - 1, P(N) is updated to P(0).

[0179] S1012 to S1016 are similar to S401 to S406 in the first embodiment.

[0180] In S1017, the synchronization circuit 311 determines whether the current main scanning direction is the forward direction or the return direction. If it is the forward direction, the synchronization circuit 311 outputs a sub-scanning trigger signal and a height scanning trigger signal to execute S1018 in parallel with S1019 and S1020. If it is the return direction, the process proceeds to S1021.

[0181] In S1018, the CPU 301 performs sub-scanning simultaneously with the main scanning in the return direction. Specifically, the CPU 301 instructs the motor control circuit 308 to move the array board 101 in the sub-scanning direction by Yp. Here, the number of pulses output from the motor control circuit 308 to the motor driver 111 is Yp / My, where My is the displacement of the array board 101 when one pulse of the voltage pulse signal is sent to the motor driver 111. In this embodiment, My = 2um.

[0182] Here, the sub-scanning is performed simultaneously with the main scanning in the return direction, thereby generating a substantially linearly inclined trajectory that intersects both the X direction and the Y direction, such as Figure 9 the trajectory 902 in. It should be noted that the moving distance of the component in the Y direction is Yp.

[0183] Based on an instruction from the CPU 301, the motor control circuit 308 outputs a pulse signal to the motor driver 111 at a speed that allows the completion of sub-scanning when the optical guiding unit 104 moves in the return direction of the main scanning direction.

[0184] S1019 is similar to the processing of S417 in the first embodiment.

[0185] In S1020, height scanning is performed to obtain focusing optical information. Specifically, the CPU 301 instructs the motor control circuit 309 to move the array board 101 by Zp in the moving direction Dir. When the value of Dir is positive, the array board 101 moves upward, and when the value of Dir is negative, the array board 101 moves downward.

[0186] The number of pulses output from the motor control circuit 309 to the motor driver 111 is Zp / Mz. Based on an instruction from the CPU 301, the motor control circuit 309 outputs a pulse signal to the motor driver 114 at a speed that allows the completion of height scanning when the optical guiding unit 104 moves in the return direction of the main scanning direction. Therefore, height scanning is performed while performing sub-scanning and while performing the main scanning in the return direction. In other words, height scanning is performed in parallel with the main scanning and sub-scanning in the return direction. On the other hand, height scanning is not performed while performing the main scanning in the forward direction.

[0187] When the sub-scanning in S1018 and the height scanning in S1020 are completed, the process proceeds to S1021.

[0188] S1021 is similar to the processing of S419 in the first embodiment.

[0189] As described above, the sample measurement device 100 according to the present embodiment acquires fluorescence signal data only during the main scan in the forward direction, and performs a sub-scan and a height scan during the main scan in the return direction without acquiring fluorescence signal data. Therefore, although the imaging time is doubled, since the main scan direction during the data acquisition period can be aligned, the influence of the position and angle errors of the optical guiding unit 104 in the forward path and the return path can be reduced.

[0190] In addition, in the present embodiment, since the time spent on the main scan in the return direction can be allocated to the sub-scan and the height scan, the speeds of the sub-scan and the height scan can be reduced. Therefore, the influence of the residual vibration caused by the sub-scan and the height scan can be reduced, which can improve the quality of the fluorescence image.

[0191] It should be noted that although the present embodiment describes the case where fluorescence signal data is acquired only during the main scan in the forward direction, fluorescence signal data can be acquired only during the main scan in the return direction, and a sub-scan and a height scan can be performed during the main scan in the forward direction without acquiring fluorescence signal data. This can be achieved by reversing the forward direction and the return direction in the determination in S1008 and S1017 described above.

[0192] It should be noted that although the present embodiment describes the operations in the imaging process, sampling and height scanning can also be performed only in one of the forward direction and the return direction in the process of acquiring height information. In this case, as in the imaging process, the influence of the position and angle errors of the optical guiding unit 104 can be reduced, and the influence of the residual vibration caused by the height scan can also be reduced, which can improve the accuracy of the acquired height information.

[0193] <Third Embodiment>

[0194] The third embodiment is different from the first embodiment in that the encoder 108 for measuring the position of the optical guiding unit 104 in the main scan direction is not included. In the first embodiment, the coordinate calculation circuit 310 calculates the position of the optical guiding unit 104 based on the signal from the encoder 108. In the present embodiment, the position of the optical guiding unit 104 is calculated based on the motor drive pulse signal from the motor control circuit 307. It should be noted that the configurations of the sample measurement device 100 and the array plate 101 in the present embodiment are similar to those in Figure 1 、 Figure 2A and Figure 2B and thus the descriptions thereof will be omitted.

[0195] Figure 11FIG. 0 is a block diagram showing the internal configuration of the controller 1116 in the third embodiment. The configuration of the controller 1116 is similar to that in the first embodiment, except that the coordinate calculation circuit 310 is replaced by the coordinate calculation circuit 1310 and the encoder 108 is not included.

[0196] The coordinate calculation circuit 1310 is a circuit that calculates the position of the light guide unit 104 based on the drive pulse voltage from the motor control circuit 307.

[0197] Figure 12 FIG. 7 is a diagram showing the operation of the piston crank mechanism.

[0198] The position x of the light guide unit 104 is represented by the following (Equation 6), where r is the length of the crank 118 of the piston crank mechanism, l is the length of the connecting rod 119 of the piston crank mechanism, and θ is the angle of the pulse motor 107.

[0199] [Formula 1]

[0200]

[0201] By multiplying the rotation angle per pulse by the number of drive pulses, θ can be calculated. Since r and l are known values, the coordinates of the light guide unit 104 are calculated using (Equation 6) each time a drive pulse is input. However, since the rotation angle per pulse is in 0.72° increments, the resulting value of x will also be discrete. Therefore, the coordinate calculation circuit 1310 estimates the coordinates between pulses by interpolating using the angular velocity.

[0202] Figure 13 FIG. 23 is a flowchart showing the operation performed by the coordinate calculation circuit.

[0203] In S1301, the coordinate calculation circuit 1310 sets the values of the internal pulse counter and the time counter to 0.

[0204] In S1302, the coordinate calculation circuit 1310 determines whether 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.

[0205] In S1303, the coordinate calculation circuit 1310 determines whether the motor has completed one rotation. For example, when the rotation angle θp of the pulse motor 107 per pulse is 0.72°, the motor completes one full rotation using 500 pulses. Therefore, when the current value Cp of the pulse counter is 499, it is determined that one full rotation has been completed, and the process proceeds to S1310. On the other hand, when the current value Cp of the pulse counter is 498 or less, it is determined that one full rotation has not been completed, and the process proceeds to S1304.

[0206] In S1304, the coordinate calculation circuit 1310 increments the pulse counter by 1.

[0207] In S1305, the coordinate calculation circuit 1310 calculates the angular velocity w by dividing the value of the time counter by the clock period. The angular velocity w is calculated using w = θp × T / Ct, where Ct is the value of the time counter and T is the clock period. However, when the value Ct of the time counter is 0, the angular velocity w is calculated as 0.

[0208] In S1306, the coordinate calculation circuit 1310 sets the time counter to 0.

[0209] In S1307, the coordinate calculation circuit 1310 calculates the angle θ. The angle θ is calculated using θ = Cp × θp + w × Ct × T, where Cp is the value of the pulse counter.

[0210] In S1308, the coordinate calculation circuit 1310 calculates the x coordinate. Specifically, the calculated angle θ is substituted into (Equation 6) to calculate the x coordinate.

[0211] In S1309, the coordinate calculation circuit 1310 outputs the calculated coordinates to the synchronization circuit 311.

[0212] In S1310, the coordinate calculation circuit 1310 sets the pulse counter to 0.

[0213] In S1311, the coordinate calculation circuit 1310 increments the pulse counter by 1.

[0214] Although the operation of the coordinate calculation circuit 1310 has been described with reference to the flowchart, since the coordinate calculation circuit 1310 is implemented on a digital circuit, the coordinate calculation circuit 1310 actually performs the operations of S1301 to S1311 in each clock cycle.

[0215] As described above, in this embodiment, the position of the light guiding unit 104 is estimated from the drive pulse signal from the motor control circuit 307 to allow synchronization of main scanning, sub-scanning, height scanning, and data acquisition without using an encoder. Since the sample measurement device 100 does not include an encoder, the manufacturing cost can be reduced.

[0216] It should be noted that although this embodiment describes the case where the rotation angle θp of the pulse motor 107 per pulse is set to 0.72°, this is not a restrictive case. For example, by using a motor driver with a microstep control function in the motor control circuit 307, θp can be divided into dozens or hundreds of steps, thereby improving the accuracy of estimating the position of the light guiding unit 104.

[0217] It should be noted that although this embodiment describes the case where the coordinate calculation circuit 1310 calculates the angular velocity based on the time difference between the rising edges of the pulse signals, this is not a restrictive case. For example, the coordinate calculation circuit 1310 can be configured to operate while the pulse motor 107 rotates at a constant speed, and can calculate the angular velocity using the rotational speed Xs in the main scanning direction specified by the user.

[0218] Although the present invention has been described in detail based on the preferred embodiments of the present invention, the present invention is not limited to the specific embodiments, and various forms that do not deviate from the essence of the present invention are also included within the scope of the present invention. For example, a part of the configuration or process from one embodiment can be combined with another embodiment.

[0219] In addition, a single hardware component can perform various types of control performed by the CPU 301, the synchronization circuit 311, or the data acquisition circuit 306 described in the above embodiments. In addition, multiple hardware components (e.g., multiple processors or circuits) can share the processing of various types of control to control the entire device.

[0220] <Other Embodiments>

[0221] The present invention can also be implemented by performing the following processing. Specifically, a program configured to implement the functions of the above embodiments is supplied to a system or device via a network or various recording media, and a computer (such as a CPU or a microprocessor unit (MPU)) of the system or device reads the program code and executes the read program code. In this case, the program and the recording medium storing the program constitute the present invention.

[0222] In addition, the disclosure of this embodiment includes the following configurations.

[0223] (Configuration 1)

[0224] A scanning device configured to scan an observation optical system above an array plate, the array plate including a plurality of spots on one surface, the scanning device comprising: an observation optical system configured to radiate primary light toward the one surface to obtain optical information related to at least a part of the plurality of spots; a scanning unit configured to perform main scanning and sub-scanning, in the main scanning, the observation optical system moves relative to the array plate in a first direction and obtains optical information, in the sub-scanning, the observation optical system moves relative to the array plate in a second direction intersecting the first direction without obtaining optical information; and an adjustment unit configured to adjust the position of the observation optical system relative to the array plate in the optical axis direction of the primary light, wherein the adjustment unit performs the adjustment when the scanning unit is in the period of sub-scanning.

[0225] (Configuration 2)

[0226] The scanning device according to Configuration 1, wherein when the scanning unit is in the period of main scanning, the adjustment unit does not perform adjustment.

[0227] (Configuration 3)

[0228] The scanning device according to Configuration 1 or 2, further comprising an image acquisition unit configured to acquire a two-dimensional image based on an output signal from the observation optical system and information on the position of the observation optical system relative to the array plate on a plane in which the observation optical system moves relative to the array plate.

[0229] (Configuration 4)

[0230] The scanning device according to any one of Configurations 1 to 3, further comprising a control unit configured to determine whether to perform adjustment based on information on a scanning sequence performed by the scanning unit.

[0231] (Configuration 5)

[0232] The scanning device according to any one of Configurations 1 to 4, further comprising a storage unit configured to store information on an imaging region defined with respect to the one surface.

[0233] (Configuration 6)

[0234] The scanning device according to any one of Configurations 1 to 5, wherein the sub-scanning includes movements corresponding to two or more movement directions on a plane in which the observation optical system moves relative to the array plate.

[0235] (Configuration 7)

[0236] The scanning device according to Configuration 6, wherein the period of the sub-scanning includes a movement in the first direction.

[0237] (Configuration 8)

[0238] The scanning device according to any one of Configurations 1 to 7 further includes an acquisition unit configured to acquire information about the array board, wherein the adjustment unit performs adjustment based on the information about the array board acquired by the acquisition unit.

[0239] (Configuration 9)

[0240] The scanning device according to Configuration 8, wherein the information about the array board includes tilt information about the array board when viewed from a first direction.

[0241] (Configuration 10)

[0242] The scanning device according to Configuration 9, wherein the tilt information about the array board is calculated based on height information about the array board acquired at at least two points.

[0243] (Configuration 11)

[0244] The scanning device according to any one of Configurations 1 to 10, wherein the scanning unit moves the observation optical system and the array board relative to each other based on information about the position of the observation optical system and information about the position of the array board.

[0245] (Configuration 12)

[0246] The scanning device according to any one of Configurations 1 to 11, wherein the adjustment unit performs adjustment based on information about the imaging region for acquiring optical information and information about the position of the observation optical system.

[0247] (Configuration 13)

[0248] The scanning device according to Configuration 12, wherein the adjustment unit performs adjustment when the position of the observation optical system is outside the imaging region.

[0249] (Configuration 14)

[0250] The scanning device according to Configuration 12 or 13, wherein the information about the imaging region is information pre-input by the user.

[0251] (Configuration 15)

[0252] The scanning device according to any one of Configurations 11 to 14 further includes a measurement unit configured to measure the position of the observation optical system, wherein the information about the position of the observation optical system is information acquired based on the position of the observation optical system measured by the measurement unit.

[0253] (Configuration 16)

[0254] The scanning device according to any one of Configurations 11 to 14 further includes a driving unit configured to move the observation optical system relative to the array plate, wherein information about the position of the observation optical system is information obtained based on a signal for driving the driving unit.

[0255] (Configuration 17)

[0256] The scanning device according to any one of Configurations 1 to 16, wherein the second direction is a direction perpendicular to the first direction.

[0257] (Configuration 18)

[0258] The scanning device according to any one of Configurations 1 to 16, wherein the second direction is a direction that is not perpendicular to the first direction but is inclined with respect to the first direction.

[0259] (Configuration 19)

[0260] The scanning device according to any one of Configurations 1 to 16, wherein the array plate has a rectangular shape when viewed from the one surface, the rectangular shape having a short side and a long side, wherein the first direction is a direction parallel to the short side of the array plate, and wherein the second direction is a direction parallel to the long side of the array plate.

[0261] (Configuration 20)

[0262] The scanning device according to any one of Configurations 1 to 16, wherein the array plate has a rectangular shape when viewed from the one surface, the rectangular shape having a short side and a long side, wherein the first direction is a direction parallel to the short side of the array plate, and wherein the second direction is a direction intersecting both the short side and the long side of the array plate.

[0263] (Configuration 21)

[0264] The scanning device according to any one of Configurations 1 to 20, wherein the observation optical system is configured to radiate light once toward at least a part of the plurality of spots and capture secondary light from at least the part of the plurality of spots.

[0265] (Method 1)

[0266] A scanning method for causing an observation optical system to scan above an array plate having a plurality of spots on one surface, the method including performing a main scan and a sub-scan and adjusting the position of the observation optical system relative to the array plate in the optical axis direction of the primary light. In the main scan, the observation optical system moves relative to the array plate in a first direction and obtains optical information related to at least a part of the plurality of spots. The observation optical system is configured to radiate primary light toward the one surface to obtain optical information. In the sub-scan, the observation optical system moves relative to the array plate in a second direction intersecting the first direction without obtaining optical information, wherein the adjustment is performed during the period of the sub-scan performed by scanning.

[0267] (Method 2)

[0268] The scanning method according to Method 1 further includes pre-acquiring information about the array plate before scanning, wherein the adjustment is performed based on the acquired information about the array plate.

[0269] (Program 1)

[0270] A program for causing a computer to execute the scanning method according to Method 1.

[0271] The present invention is not limited to the above embodiments, and various changes and modifications can be made without departing from the spirit and scope of the present invention. Therefore, the following claims are appended to define the scope of the present invention.

[0272] This application claims priority based on Japanese Patent Application No. 2022-189179 filed on November 28, 2022, and incorporates the entire content thereof herein by reference.

[0273] 100 Sample measurement device (scanning device)

[0274] 101 Array plate

[0275] 104 Light guiding unit

[0276] 105 Light sensor

[0277] 106 Piston crank mechanism

[0278] 107 Pulse motor

[0279] 109 Linear stage

[0280] 110 Pulse motor

[0281] 112 Linear stage

[0282] 113 Pulse motor

Claims

1. A scanning device configured to scan an observation optical system above an array plate, the array plate including a plurality of spots on one surface, the scanning device comprising: An observation optical system configured to radiate primary light toward the one surface to obtain optical information related to at least a part of the plurality of spots; A scanning unit configured to perform main scanning and sub-scanning. In the main scanning, the observation optical system moves relative to the array plate in a first direction and obtains the optical information. In the sub-scanning, the observation optical system moves relative to the array plate in a second direction intersecting the first direction without obtaining the optical information; And An adjustment unit configured to adjust the position of the observation optical system relative to the array plate in the optical axis direction of the primary light, wherein the adjustment unit performs the adjustment when the scanning unit is in the period of the sub-scanning.

2. The scanning device according to claim 1, wherein when the scanning unit is in the period of the main scanning, the adjustment unit does not perform the adjustment.

3. The scanning device according to claim 1, further comprising an image acquisition unit configured to acquire a two-dimensional image based on an output signal from the observation optical system and information about the position of the observation optical system relative to the array plate on a plane where the observation optical system moves relative to the array plate.

4. The scanning device according to claim 1, further comprising a control unit configured to determine whether to perform the adjustment based on information about a scanning sequence executed by the scanning unit.

5. The scanning device according to claim 1, further comprising a storage unit configured to store information about an imaging region defined relative to the one surface.

6. The scanning device according to claim 1, wherein the sub-scanning includes movements corresponding to two or more movement directions on a plane where the observation optical system moves relative to the array plate.

7. The scanning device according to claim 6, wherein the sub-scanning includes a movement in the first direction.

8. The scanning device according to claim 1 or 2, further comprising an acquisition unit configured to acquire information about the array plate, wherein the adjustment unit performs the adjustment based on the information about the array plate acquired by the acquisition unit.

9. The scanning device according to claim 8, wherein the information about the array plate includes tilt information about the array plate when observed from the first direction.

10. The scanning device according to claim 8, wherein the tilt information about the array plate is calculated based on height information about the array plate acquired at at least two points.

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

12. The scanning device according to claim 1, wherein the adjustment unit performs the adjustment based on information about the imaging region where the optical information is acquired and information about the position of the observation optical system.

13. The scanning device according to claim 12, wherein the adjustment unit performs the adjustment when the position of the observation optical system is outside the imaging region.

14. The scanning device according to claim 12 or 13, wherein the information about the imaging region is information pre-input by a user.

15. The scanning device according to claim 11 or 12, further comprising a measurement unit configured to measure the position of the observation optical system, wherein the information about the position of the observation optical system is information acquired based on the position of the observation optical system measured by the measurement unit.

16. The scanning device according to claim 11 or 12, further comprising a drive unit configured to move the observation optical system relative to the array plate, wherein the information about the position of the observation optical system is information acquired based on a signal for driving the drive unit.

17. The scanning device according to claim 1 or 2, wherein the second direction is a direction perpendicular to the first direction.

18. The scanning device according to claim 1 or 2, wherein the second direction is a direction that is not perpendicular to the first direction but is inclined with respect to the first direction.

19. The scanning device according to claim 1 or 2, wherein the array plate has a rectangular shape when viewed from the one surface, the rectangular shape having a short side and a long side, wherein the first direction is a direction parallel to the short side of the array plate, and wherein the second direction is a direction parallel to the long side of the array plate.

20. The scanning device according to claim 1 or 2, wherein the array plate has a rectangular shape when viewed from the one surface, the rectangular shape having a short side and a long side, wherein the first direction is a direction parallel to the short side of the array plate, and wherein the second direction is a direction intersecting both the short side and the long side of the array plate.

21. The scanning device according to claim 1 or 2, wherein the observation optical system is configured to irradiate the primary light toward at least a part of the plurality of spots and capture the secondary light from at least a part of the plurality of spots.

22. A scanning method for scanning an observation optical system above an array plate including a plurality of spots on one surface, the method comprising: Perform a main scan and a sub-scan. In the main scan, the observation optical system moves relative to the array plate in a first direction and acquires optical information related to at least a part of the plurality of spots. The observation optical system is configured to irradiate light once toward the one surface to acquire the optical information. In the sub-scan, the observation optical system moves relative to the array plate in a second direction intersecting the first direction without acquiring the optical information; and Adjust the position of the observation optical system relative to the array plate in the optical axis direction of the once light, wherein the adjustment is performed during the sub-scan performed by the scan.

23. The scanning method according to claim 22, further comprising pre-acquiring information about the array plate before the scan, wherein the adjustment is performed based on the acquired information about the array plate.

24. A program for causing a computer to execute the scanning method according to claim 22 or 23.

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