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WO2018188440A1 - Procédé, dispositif et système d'imagerie - Google Patents

Procédé, dispositif et système d'imagerie Download PDF

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Publication number
WO2018188440A1
WO2018188440A1 PCT/CN2018/078799 CN2018078799W WO2018188440A1 WO 2018188440 A1 WO2018188440 A1 WO 2018188440A1 CN 2018078799 W CN2018078799 W CN 2018078799W WO 2018188440 A1 WO2018188440 A1 WO 2018188440A1
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WO
WIPO (PCT)
Prior art keywords
sample
lens module
light
light intensity
sharpness value
Prior art date
Application number
PCT/CN2018/078799
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English (en)
Chinese (zh)
Inventor
孙瑞涛
徐剑峰
金欢
徐家宏
周志良
姜泽飞
颜钦
Original Assignee
深圳市瀚海基因生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Publication date
Priority claimed from CN201710227964.3A external-priority patent/CN108693625B/zh
Priority claimed from CN201710227938.0A external-priority patent/CN108693113B/zh
Priority claimed from CN201710227936.1A external-priority patent/CN108693624B/zh
Application filed by 深圳市瀚海基因生物科技有限公司 filed Critical 深圳市瀚海基因生物科技有限公司
Publication of WO2018188440A1 publication Critical patent/WO2018188440A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes

Definitions

  • the present invention relates to the field of optical detection, and more particularly to an imaging method, apparatus and system.
  • Sequencing includes the determination of nucleic acid sequences.
  • the current sequencing platforms on the market include a generation of sequencing platforms, second-generation sequencing platforms and three generations of sequencing platforms.
  • the sequencing instrument includes a detection module that utilizes the detection module to transform and/or collect information changes produced by biochemical reactions in the sequence determination to determine the sequence.
  • the detection module generally includes an optical detection module, a current detection module, and an acid-base (pH) detection module.
  • the sequencing platform based on the principle of optical detection performs sequence determination by analyzing the changes in the optical signals in the detected sequencing biochemical reactions.
  • the optical detection system with auto-focus module is equipped with a matching focus control program, which can be directly called and controlled. It is easy to use, but often does not sell the auto-focus module separately. Buyers can buy the whole system together and have high cost.
  • embodiments of the present invention aim to at least solve one of the technical problems existing in the related art or at least provide an alternative practical solution. To this end, embodiments of the present invention are required to provide an imaging method, an optical detection system, and a control device.
  • Embodiments of the present invention provide an imaging method for an optical detection system, the optical detection system including an imaging device and a stage, the imaging device including a lens module and a focus module, the lens module including An optical axis, the stage for carrying a sample, the method comprising the step of focusing: using the focusing module to emit light onto a sample placed on the stage; causing the lens module to follow the light Moving the axis to the first set position; moving the lens module from the first set position to the sample along the optical axis at a first set step and determining whether the focus module receives The light reflected by the sample; when the focusing module receives the light reflected by the sample, causing the lens module to have a second setting smaller than the first set step Moving a step along the optical axis and performing image acquisition on the sample by using the imaging device, and determining whether a sharpness value of the image collected by the imaging device reaches a set threshold; When the sharpness value reaches the set threshold, Keep the current position of the lens module as the location.
  • This method is particularly suitable for devices that include a precision optical system that is difficult to find a clear plane, such as optical inspection equipment with high magnification lenses. In this way, the cost can be reduced.
  • An optical detection system includes a control device, an imaging device, and a stage, the imaging device includes a lens module and a focus module, the lens module includes an optical axis, and the carrier is configured to carry a sample, the control device is configured to: use the focusing module to emit light onto a sample placed on the stage; and move the lens module along the optical axis to a first set position; The lens module moves from the first set position to the sample along the optical axis at a first set step and determines whether the focus module receives the light reflected by the sample; When the focusing module receives the light reflected by the sample, moving the lens module along the optical axis by using a second set step smaller than the first set step size The imaging device performs image acquisition on the sample, and determines whether the sharpness value of the image collected by the imaging device reaches a set threshold; when the sharpness value of the image reaches the set threshold, Saving the current position of the lens module as a save location .
  • the optical inspection system is especially suitable for devices that contain precise optical systems that are difficult to find clear planes, such as optical inspection equipment with high magnification lenses. In this way, the cost can be reduced.
  • a control device for controlling imaging is used in an optical detection system, the optical detection system includes an imaging device and a focus module, and the control device includes: a storage device for storing data, The data includes a computer executable program; a processor for executing the computer executable program, and executing the computer executable program includes the method of performing the above embodiments.
  • a computer readable storage medium for storing a program for execution by a computer, and executing the program includes performing the above method.
  • the computer readable storage medium may include read only memory, random access memory, magnetic or optical disks, and the like.
  • FIG. 1 is a schematic flow chart of an image forming method according to an embodiment of the present invention.
  • FIG. 2 is a schematic view showing the positional relationship between a lens module and a sample according to an embodiment of the present invention.
  • FIG 3 is a partial structural schematic view of an optical detecting system according to an embodiment of the present invention.
  • FIG 4 is another schematic flow chart of an imaging method according to an embodiment of the present invention.
  • FIG. 5 is a schematic flow chart of still another embodiment of the imaging method according to the embodiment of the present invention.
  • FIG. 6 is a block diagram of an optical detection system in accordance with an embodiment of the present invention.
  • first and second are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated.
  • features defining “first” or “second” may include one or more of the described features either explicitly or implicitly.
  • the meaning of "a plurality" is two or more unless specifically and specifically defined otherwise.
  • connection should be understood broadly, for example, it may be a fixed connection, a detachable connection, or an integral connection;
  • the mechanical connections may also be electrical connections or may communicate with each other; they may be directly connected or indirectly connected through an intermediate medium, and may be internal communication of two elements or an interaction relationship of two elements.
  • specific meanings of the above terms in the present invention can be understood on a case-by-case basis.
  • the term "invariant”, for example, relating to distance, object distance, and/or relative position, etc., may be expressed as a numerical value, a numerical range, or a change in quantity, which may be absolutely constant or may be Relatively constant, the so-called relative constant is to maintain a certain range of deviation or a preset acceptable range. Unless otherwise stated, "invariant" involving distance, object distance, and/or relative position is relatively constant.
  • Sequence determination as used in the context of the present invention is the same as nucleic acid sequence determination, including DNA sequencing and/or RNA sequencing, including long fragment sequencing and/or short fragment sequencing.
  • sequence determination reaction is the same as the sequencing reaction.
  • an embodiment of the present invention provides an imaging method for an optical detection system
  • the optical detection system includes an imaging device 102 and a stage
  • the imaging device 102 includes a lens module 104 and a focus module 106.
  • the lens module 104 includes an optical axis OP for carrying the sample 300.
  • the imaging method includes the following focusing steps: S11, using the focusing module 106 to emit light onto the sample 300 placed on the stage; S12, moving the lens module 104 along the optical axis OP to the first set position; S13, making the lens
  • the module 104 moves from the first set position to the sample 300 along the optical axis 300 in a first set step and determines whether the focus module 106 receives the light reflected by the sample 300; the focus module 106 receives the sample 300 In the reflected light, S14, the lens module 104 is moved along the optical axis OP by a second set step smaller than the first set step and the image is collected by the imaging device 102, and the imaging device 102 is determined.
  • the sharpness value of the collected image reaches the set threshold; when the sharpness value of the image reaches the set threshold, S15, the current position of the lens module 104 is saved as the save position.
  • This method is particularly suitable for devices that include a precision optical system that is difficult to find a clear plane, such as optical inspection equipment with high magnification lenses.
  • the sample 300 includes a carrying device 200 and a sample 302 to be tested located in the carrying device, and the sample 302 to be tested is a biomolecule, such as a nucleic acid, and the lens module 104 is located in the carrying device.
  • the carrying device 200 has a front panel 202 and a rear panel (lower panel), each panel having two surfaces, and the sample to be tested 302 is connected to the upper surface of the lower panel, that is, the sample to be tested 302 is located below the lower surface 204 of the front panel 202.
  • the imaging device 102 is an image for collecting the sample 302 to be tested, and the sample to be tested 302 is located below the lower surface 204 of the front panel 202 of the carrier device 200, the movement of the lens module 104 at the beginning of the focusing process. It is to find the medium interface 204 where the sample 302 to be tested is located to improve the success rate of the imaging device 102 to acquire a clear image.
  • the sample 302 to be tested is a solution
  • the front panel 202 of the carrying device 200 is glass
  • the medium interface 204 of the carrying device 200 and the sample to be tested 302 is the lower surface 204 of the front panel 202 of the carrying device 200. That is, the interface between the glass and the liquid medium.
  • the sample to be tested 302 of the imaging device 102 is required to be located below the lower surface 204 of the front panel 202. At this time, the image captured by the imaging device 102 is used to discriminate the clear surface for finding the sample 302 to be tested clearly. Can be called focus.
  • the front panel 202 of the sample 302 to be tested has a thickness of 0.175 mm.
  • the carrier device 200 can be a slide, the sample 302 to be tested is placed on the slide, or the sample 302 to be tested is sandwiched between the two slides.
  • the carrier device 200 can be a reaction device, for example, a sandwich-like chip carrying a panel above and below, and the sample 302 to be tested is disposed on the chip.
  • the imaging device 102 includes a microscope 107 and a camera 108.
  • the lens module 104 includes an objective lens 110 of the microscope and a lens module 112 of the camera 108.
  • the focus module 106 can pass the dichroic color separation.
  • the dichroic beam splitter is fixed to the lens module 112 of the camera 108, and the dichroic beam splitter 114 is located between the lens module 112 of the camera 108 and the objective lens 110.
  • the dichroic beam splitter 114 includes a dual c-mount splitter.
  • the dichroic beam splitter 114 can reflect the light emitted by the focusing module 106 to the objective lens 110 and can pass visible light through the lens module 112 of the camera 108 into the camera 108, as shown in FIG.
  • the movement of the lens module 104 may refer to the movement of the objective lens 110, and the position of the lens module 104 may refer to the position of the objective lens 110. In other embodiments, other lenses of the lens module 104 can be selected to achieve focus.
  • the microscope 107 further includes a barrel lens 111 between the objective lens 110 and the camera 108.
  • the stage can move the sample 200 in a plane perpendicular to the optical axis OP (eg, the Z-axis) of the lens module 104 (eg, the XY plane), and/or can drive the sample 300 along the lens module.
  • the optical axis OP of 104 (such as the Z axis) moves.
  • the plane that the stage drives the sample 300 to move is not perpendicular to the optical axis OP, ie, the plane of motion of the sample is at an angle other than zero to the XY plane, and the imaging method is still applicable.
  • the imaging device 102 can also drive the objective lens 110 to move along the optical axis OP of the lens module 104 to perform focusing.
  • the imaging device 102 drives the objective lens 110 to move using a drive such as a stepper motor or a voice coil motor.
  • the positions of the objective lens 110, the stage, and the sample 300 may be set on the negative axis of the Z axis, and the first set position may be the Z axis.
  • the coordinate position on the negative axis It can be understood that, in other embodiments, the relationship between the coordinate system and the camera and the objective lens 110 may be adjusted according to actual conditions, and is not specifically limited herein.
  • the first set step size S1 is more suitable, because S1 is too large to cross an acceptable focus range, and S1 is too small to increase the time overhead.
  • the lens module 104 is caused to continue moving toward the sample 300 along the optical axis OP at the first set step.
  • the lens module 104 when the sharpness value of the image does not reach the set threshold, the lens module 104 is caused to continue moving along the optical axis OP by the second set step.
  • the optical detection system can be applied to a sequencing system, or the sequencing system includes an optical detection system.
  • the lens module 104 when the lens module 104 moves, determining whether the current position of the lens module 104 exceeds the second set position; when the current position of the lens module 104 exceeds the second set position, stopping the moving lens
  • the module 104 either performs a focusing step.
  • the first set position and the second set position can limit the range of movement of the lens module 104, so that the lens module 104 can stop moving when the focus cannot be successfully performed, thereby avoiding waste of resources or damage of the device, or
  • the lens module 104 refocuses when the focus cannot be successfully achieved, thereby improving the automation of the imaging method.
  • the settings are adjusted such that the range of motion of the lens module 104 is as small as possible to meet the implementation of the solution.
  • the range of movement of the lens module 104 can be set to 200 ⁇ m ⁇ 10 ⁇ m or [190 ⁇ m, 250 ⁇ m] according to optical path characteristics and experience.
  • another set position may be determined depending on the determined range of movement and the setting of any of the second set position and the first set position.
  • the second set position is set to be the lowest position of the upper surface 205 of the front panel 202 of the reaction device 200 to the next depth of field, and the movement range of the lens module 104 is set to 250 ⁇ m.
  • the setting position is confirmed.
  • the coordinate position corresponding to the position of the next depth of field is a position that becomes smaller in the negative direction of the Z axis.
  • the movement range is a section on the negative axis of the Z-axis.
  • the first set position is nearlimit
  • the second set position is farlimit
  • the coordinate positions corresponding to nearlimit and farlimit are on the negative axis of the Z axis
  • nearlimit -6000um
  • farlimit -6350um
  • the range of movement defined between nearlimit and farlimit is 350um. Therefore, when the coordinate position corresponding to the current position of the lens module 104 is smaller than the coordinate position corresponding to the second set position, it is determined that the current position of the lens module 104 exceeds the second set position.
  • the position of farlimit is the position of the next depth L of the lowermost surface 205 of the front panel 202 of the reaction apparatus 200.
  • the depth of field L is the depth of field of the lens module 104.
  • the coordinate positions corresponding to the first set position and/or the second set position may be specifically set according to actual conditions, and are not specifically limited herein.
  • the focus module 106 includes a light source 116 for emitting light onto the sample 300 and a light sensor 118 for receiving light reflected by the sample 300. In this way, the illumination of the focus module 106 and the reception of light can be achieved.
  • the light source 116 can be an infrared light source 116, and the light sensor 118 can be a photo diode.
  • the infrared light emitted by the light source 116 enters the objective lens 110 through the reflection of the dichroic beam splitter and is projected through the objective lens 110 to the sample 300.
  • the sample 300 can reflect infrared light projected through the objective lens 110.
  • the sample 300 includes the carrier device 200 and the sample 302 to be tested, the light reflected by the received sample 300 is light reflected by the lower surface 204 of the front panel of the carrier device 200.
  • the distance between the objective lens 110 and the sample 300 is in an optical imaging suitable range, and can be used for imaging of the imaging device 102. In one example, the distance is 20-40um.
  • the lens module 104 is moved at a second set step size smaller than the first set step, so that the optical detecting system can find the optimal imaging position of the lens module 104 in a smaller range.
  • the sharpness value of the image can be used as an evaluation value for image focus. In one embodiment, it is determined whether the sharpness value of the image acquired by the imaging device 102 reaches a set threshold value that can be passed through the image processing hill climbing algorithm. It is determined whether the sharpness value reaches the maximum value at the peak of the sharpness value by calculating the sharpness value of the image output by the imaging device 102 at each position of the objective lens 110, thereby determining whether the lens module 104 reaches the imaging device 102 during imaging. The location of the clear face. It can be understood that in other embodiments, other image processing algorithms may also be utilized to determine whether the sharpness value reaches the maximum value at the peak.
  • the sharpness value of the image reaches the set threshold, the current position of the lens module 104 is saved as the storage position, so that the imaging device 102 can output a clear image when the sequence measurement reaction is taken.
  • the method further includes the step of: S16, causing the lens module 104 to be smaller than the first set step and greater than the first
  • the third set step of the second step is moved along the optical axis OP to the sample 300, and the first light intensity parameter is calculated according to the light intensity of the light received by the focus module 106, and it is determined whether the first light intensity parameter is greater than The first set light intensity threshold; when the first light intensity parameter is greater than the first set light intensity threshold, step S14 is performed.
  • step S14 is performed.
  • the lens module 104 is caused to continue moving toward the sample 300 along the optical axis OP in the third set step.
  • the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the first light intensity parameter being the light of the light received by the two light sensors 118. Strong average. As such, the first light intensity parameter is calculated by the average of the light intensities of the light received by the two light sensors 118 such that the weak light signal is excluded from the more accurate.
  • the first set light intensity threshold nSum 40.
  • the third set step size S2 0.005 mm. It can be understood that, in other examples, the third set step size may also adopt other values, which are not specifically limited herein.
  • the method further includes the following steps: S16, the lens module 104 is made smaller than the first set step and greater than The third set step of the second set step moves to the sample 300 along the optical axis OP, and calculates a first light intensity parameter according to the light intensity of the light received by the focus module 106, and determines whether the first light intensity parameter is And greater than the first set light intensity threshold; when the first light intensity parameter is greater than the first set light intensity threshold, S17, the lens module 104 is made smaller than the third set step and greater than the second set step The fourth set step moves along the optical axis OP to the sample 300, and calculates a second light intensity parameter according to the light intensity of the light received by the focus module 106, and determines whether the second light intensity parameter is smaller than the second set light intensity threshold.
  • step S14 is performed.
  • the second light intensity parameter is less than the second set light intensity threshold.
  • the comparison of the second set light intensity threshold can eliminate the strong reflected light signal at the non-media interface position, such as the interference of the oil/reflected light signal of the objective lens 110 on the focus/focus.
  • the lens module 104 is caused to continue moving toward the sample 300 along the optical axis OP in the third set step.
  • the lens module 104 is caused to continue moving toward the sample 300 along the optical axis OP in the fourth set step.
  • the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the first light intensity parameter being the light of the light received by the two light sensors 118.
  • the strong average value, the light intensity of the light received by the two light sensors 118 has a first difference, and the second light intensity parameter is the difference between the first difference and the set compensation value.
  • the second light intensity parameter is calculated by the light intensity of the light received by the two light sensors 118 such that the optical signal that excludes strong reflection is more accurate.
  • the first set light intensity threshold nSum 40.
  • the lens module 104 when the lens module 104 is moved by the second set step, it is determined whether the first sharpness value of the pattern corresponding to the current position of the lens module 104 is greater than the previous one of the lens module 104. a second sharpness value of the image corresponding to the position; when the first sharpness value is greater than the second sharpness value and the sharpness difference between the first sharpness value and the second sharpness value is greater than the set difference value, Having the lens module 104 continue to move toward the sample 300 along the optical axis OP in a second set step; at a sharpness between the first sharpness value and the second sharpness value and between the first sharpness value and the second sharpness value When the difference value is less than the set difference value, the lens module 104 continues to move along the optical axis OP to the sample 300 at a fifth set step size smaller than the second set step size to cause the image captured by the imaging device 102.
  • the sharpness value reaches a set threshold; when the second sharpness value is greater than the first sharpness value and the sharpness difference between the second sharpness value and the first sharpness value is greater than the set difference, the lens module is made 104 moves away from the sample 300 along the optical axis OP in a second set step; the second sharpness value is greater than the first sharpness value and the second sharpness value and the first sharpness value are When the sharpness difference is less than the set difference, the lens module 104 is moved away from the sample 300 along the optical axis OP by the fifth set step to make the sharpness value of the image collected by the imaging device 102 reach the set threshold. . In this way, the position of the lens module 104 corresponding to the peak of the sharpness value can be accurately found, so that the image output by the imaging device is clear.
  • the second set step size can be used as the coarse adjustment step Z1
  • the fifth set step size can be used as the fine adjustment step length Z2
  • the coarse adjustment range Z3 can be set.
  • the setting of the coarse adjustment range Z3 can stop the movement of the lens module 104 when the sharpness value of the image cannot reach the set threshold, thereby saving resources.
  • the coarse adjustment range Z3 is the adjustment range, that is, the adjustment range on the Z axis is (T, T+Z3).
  • the lens module 104 is moved in the first direction (such as the direction in which the optical axis OP approaches the sample 300) in the range of (T, T+Z3) by the step size Z1, and compared with the current position of the lens module 104.
  • R1>R2 and R1-R2>R0 it means that the sharpness value of the image is close to the set threshold and farther from the set threshold, so that the lens module 104 continues to move in the first direction by the step size Z1 to quickly The ground is close to the set threshold.
  • R1>R2 and R1-R2 ⁇ R0 it means that the sharpness value of the image is close to the set threshold and is closer to the set threshold, so that the lens module 104 moves in the first direction by the step size Z2, so as to be smaller.
  • the step size is close to the set threshold.
  • R2>R1 and R2-R1>R0 it means that the sharpness value of the image has crossed the set threshold and is far from the set threshold, so that the lens module 104 has the step size Z1 in the opposite direction to the first direction.
  • the two directions e.g., in the direction away from the sample 300 along the optical axis OP) move to quickly approach the set threshold.
  • the fifth set step size can be adjusted to accommodate the step size when approaching the set threshold is not too large or too small.
  • the method further includes the following focusing step: acquiring the relative position of the lens module 104 and the sample 300 when the lens module 104 is in the storage position; and controlling the lens module when the sample 300 is moved by the loading platform Movement of 104 to maintain relative position. In this way, it can be ensured that the image captured by the imaging device 102 at different positions of the sample 300 is kept clear and the focus is achieved.
  • the sample 300 is tilted due to physical errors of the stage and/or the sample 300. Therefore, when the sample 300 is moved by the stage, the distance between the different positions of the surface of the sample 300 and the lens module 104 may occur. Variety. Therefore, when the sample 300 is moved relative to the optical axis OP of the lens module 104, the imaging position of the imaging device 102 by the imaging device 102 is maintained at the clear surface position. This process is called chasing.
  • the sample 300 is moved by the stage, including the sample 300 moving along the X1 axis parallel to the X axis, and the sample 300 moving along the Y1 axis parallel to the Y axis, and the sample 300 moving along the plane X1Y1 defined by the X1 axis and the Y1 axis, and The sample 300 moves along the tilted X axis, and the sample 300 moves along the tilted Y axis, and the sample 300 moves along a plane XY defined oblique to the X and Y axes.
  • the stage when the sample 300 is moved by the stage, it is determined whether the current position of the lens module 104 exceeds the third set position; when the current position of the lens module 104 exceeds the third set position, the load is utilized.
  • the stage drives the sample 300 to move along the optical axis OP and performs a focusing step; when the number of movements reaches the set number of times and the current position of the lens module 104 still exceeds the third set position, it is determined that the tracking failure has failed. In this way, the limitation of the third set position and the number of movements enables the lens module 104 to perform refocusing when the focus recovery fails.
  • the third set position may be nPos
  • the coordinate position corresponding to nPos is on the negative axis of the Z axis
  • the coordinate position corresponding to nPos is greater than the coordinate position corresponding to the second set position farlimit.
  • refocusing is performed to adjust the position of the lens module 104 to attempt successful tracking.
  • the process of chasing the focus if the number of times the lens module 104 is moved reaches the set number of times, the current position of the lens module 104 is still beyond the third set position, the focus cannot be recovered, the focus recovery is determined, the pause is resumed, and the focus is re-focused. Clear face.
  • the coordinate position corresponding to the third set position is an empirical value. When the value is smaller than this value, the image captured by the imaging device 102 is blurred and the probability of chasing a large probability fails.
  • the set number is an empirical value, which can be set according to the actual situation.
  • the relative position when the current position of the lens module 104 does not exceed the third set position, the relative position is determined to be unchanged.
  • the relative positions include relative distances and relative directions. Further, to simplify the operation, the relative position may refer to the relative distance, and the relative position does not mean that the object distance of the imaging system of the imaging device 102 is constant, so that different positions of the sample 300 can be clearly imaged by the imaging device.
  • an optical detection system 100 includes a control device 101, an imaging device 102, and a loading platform 103.
  • the imaging device 102 includes a lens module 104 and a focusing module 106.
  • the lens module 104 includes light.
  • the axis OP, the stage 103 is used to carry the sample 300, and the control device 101 is configured to: use the focusing module 106 to emit light onto the sample 300 placed on the stage 103; and move the lens module 104 along the optical axis OP to the first Setting the position; moving the lens module 104 from the first set position to the sample 300 along the optical axis OP by the first set step and determining whether the focus module 106 receives the light reflected by the sample 300; When receiving the light reflected by the sample 300, the lens module 104 is moved along the optical axis OP by a second set step smaller than the first set step and the image is collected by the imaging device 102, and is determined. Whether the sharpness value of the image collected by the imaging device 102 reaches the set threshold; when the sharpness value of the image reaches the set threshold, the current position of the lens module 104 is saved as the save position.
  • control device 101 includes a device having data processing and control capabilities, such as a personal computer, an embedded system, a cell phone, a tablet, a laptop, and the like.
  • the focus module 106 includes a light source 116 for emitting light onto the sample 300 and a light sensor 118 for receiving light reflected by the sample 300.
  • control device 101 can control the light source 116 to emit light and control the light sensor 118 to receive light.
  • the control device 101 when the focus module 106 receives the light reflected by the sample 300, the control device 101 is configured to: cause the lens module 104 to be smaller than the first set step and greater than the second set step.
  • the third set step moves along the optical axis OP to the sample 300, and calculates a first light intensity parameter according to the light intensity of the light received by the focus module 106, and determines whether the first light intensity parameter is greater than the first set light intensity. Threshold; when the first light intensity parameter is greater than the first set light intensity threshold, the lens module 104 is moved along the optical axis by the second set step and the image is collected by the imaging device 102, and the imaging device 102 is determined. Whether the sharpness value of the acquired image reaches the set threshold.
  • the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the first light intensity parameter being the light of the light received by the two light sensors 118. Strong average.
  • the control device 101 when the focus module 106 receives the light reflected by the sample 300, the control device 101 is configured to: cause the lens module 104 to be smaller than the first set step and greater than the second set step.
  • the third set step moves along the optical axis OP to the sample 300, and calculates a first light intensity parameter according to the light intensity of the light received by the focus module 106, and determines whether the first light intensity parameter is greater than the first set light intensity.
  • the lens module 104 when the first light intensity parameter is greater than the first set light intensity threshold, the lens module 104 is disposed along the optical axis OP with a fourth set step size that is less than the third set step size and greater than the second set step size Moving to the sample 300, and calculating a second light intensity parameter according to the light intensity of the light received by the focus module 104, determining whether the second light intensity parameter is less than a second set light intensity threshold; and the second light intensity parameter is less than the second light intensity parameter
  • the lens module 104 is moved along the optical axis OP by the second set step and the image is collected by the imaging device 102, and the sharpness of the image collected by the imaging device 102 is determined. Whether the value reaches the set threshold.
  • the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the first light intensity parameter being the light of the light received by the two light sensors 118.
  • the strong average value, the light intensity of the light received by the two light sensors 118 has a first difference, and the second light intensity parameter is the difference between the first difference and the set compensation value.
  • the control device 101 when the lens module 104 is moved by the second set step, the control device 101 is configured to determine whether the first sharpness value of the pattern corresponding to the current position of the lens module 104 is greater than the lens mode.
  • the lens module 104 is caused to continue moving along the optical axis OP to the sample 300 in a second set step; the first sharpness value is greater than the second sharpness value and the first sharpness value and the second sharpness are
  • the sharpness difference between the values is less than the set difference
  • the lens module 104 continues to move along the optical axis OP to the sample 300 at a fifth set step size that is less than the second set step size to cause the imaging device 102 to
  • the sharpness value of the collected image reaches a set threshold; when the second sharpness value is greater than the first sharpness
  • the control device 101 when the lens module 104 is moved, the control device 101 is configured to determine whether the current position of the lens module 104 exceeds a second set position, and the first set position and the second set position together define the lens.
  • the control device 101 when the control device 101 performs focusing, the focusing step in the method of the above embodiment can be performed.
  • control device 101 is configured to: determine the relative position of the lens module 104 and the sample 300 when the lens module 104 is in the storage position; and control the lens module 104 when the sample 300 is moved by the carrier 103. The movement to keep the relative position unchanged.
  • the control device 101 when the stage 103 is used to drive the sample 300 to move, the control device 101 is configured to determine whether the current position of the lens module 104 exceeds the third set position; the current position of the lens module 104 exceeds the third position.
  • the position is fixed, the sample 300 is moved by the stage 103 to perform focusing; when the number of movements of the sample 300 reaches the set number of times and the current position of the lens module 104 is still beyond the third set position, it is determined that the focus recovery has failed.
  • a control device 101 for controlling imaging is provided for an optical detection system 100.
  • the optical detection system 100 includes an imaging device 102 and a carrier 103.
  • the control device 101 includes a storage device 120.
  • the data includes a computer executable program; a processor 122 for executing a computer executable program, and executing the computer executable program includes the method of performing any of the above embodiments.
  • a computer readable storage medium for storing a program for execution by a computer, the program comprising the method of any of the above embodiments.
  • the computer readable storage medium may include read only memory, random access memory, magnetic or optical disks, and the like.
  • a "computer-readable storage medium” can be any apparatus that can contain, store, communicate, propagate, or transport a program for use in an instruction execution system, apparatus, or device, or in conjunction with such an instruction execution system, apparatus, or device.
  • computer readable storage media include the following: electrical connections (electronic devices) having one or more wires, portable computer disk cartridges (magnetic devices), random access memory (RAM) , read only memory (ROM), erasable editable read only memory (EPROM or flash memory), fiber optic devices, and portable compact disk read only memory (CDROM).
  • the computer readable storage medium may even be a paper or other suitable medium on which the program can be printed, as it may be optically scanned, for example by paper or other medium, followed by editing, interpretation or, if necessary, other Processing is performed in a suitable manner to obtain the program electronically and then stored in computer memory.
  • each functional unit in each embodiment of the present invention may be integrated into one processing module, or each unit may exist physically separately, or two or more units may be integrated into one module.
  • the above integrated modules can be implemented in the form of hardware or in the form of software functional modules.
  • the integrated modules, if implemented in the form of software functional modules and sold or used as stand-alone products, may also be stored in a computer readable storage medium.

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  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)
  • Automatic Focus Adjustment (AREA)
  • Studio Devices (AREA)

Abstract

L'invention concerne un procédé d'imagerie, destiné à être utilisé dans un système de détection optique (100). Le système de détection optique (100) comprend un dispositif d'imagerie (102) et une plateforme de support (103). Le dispositif d'imagerie (102) comprend un module de lentille (104) et un module de mise au point (106). Le module de lentille (104) comprend un axe optique. Le procédé comprend les étapes de mise au point suivantes : l'utilisation du module de mise au point (106) pour émettre de la lumière vers un échantillon (300) ; le déplacement du module de lentille (104) vers une première position de réglage le long de l'axe optique ; le déplacement du module de lentille (104) vers l'échantillon (300) dans une première taille d'étape définie le long de l'axe optique, et la détermination si le module de mise au point (106) reçoit la lumière réfléchie par l'échantillon (300) ou non ; si le module de mise au point (106) reçoit la lumière réfléchie par l'échantillon (300), le déplacement du module de lentille (104) dans une seconde taille d'étape définie, inférieure à la première taille d'étape définie, le long de l'axe optique et la réalisation d'une collecte d'image sur l'échantillon (300) à l'aide du dispositif d'imagerie (102), et la détermination si la valeur de netteté d'une image collectée par le dispositif d'imagerie (102) atteint un seuil défini ou non ; et si la valeur de netteté de l'image atteint le seuil défini, le stockage de la position actuelle du module de lentille (104) en tant que position de stockage. Le procédé d'imagerie peut assurer que l'échantillon (300) est imagé distinctement.
PCT/CN2018/078799 2017-04-10 2018-03-13 Procédé, dispositif et système d'imagerie WO2018188440A1 (fr)

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CN201710227964.3A CN108693625B (zh) 2017-04-10 2017-04-10 成像方法、装置及系统
CN201710227938.0A CN108693113B (zh) 2017-04-10 2017-04-10 成像方法、装置及系统
CN201710227936.1A CN108693624B (zh) 2017-04-10 2017-04-10 成像方法、装置及系统
CN201710227964.3 2017-04-10
CN201710227938.0 2017-04-10
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