WO2018188442A1

WO2018188442A1 - Imaging method, device and system

Info

Publication number: WO2018188442A1
Application number: PCT/CN2018/078836
Authority: WO
Inventors: 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: 2017-04-10
Filing date: 2018-03-13
Publication date: 2018-10-18
Also published as: WO2018188441A1; WO2018188440A1

Abstract

An imaging method, for use in an optical detection system (100). The optical detection system (100) comprises an imaging device (102) and a bearing platform (103). The imaging device (102) comprises a lens module (104) and a focus module (106). The method comprises the following focusing steps: using the focus module (106) to emit light to a sample (300) on the bearing platform (103); moving the lens module (104) to a first setting position along an optical axis; moving the lens module (104) from the first setting position to the sample (300) in a first set step size along the optical axis, and determining whether the focus module (106) receives the light reflected from the sample (300) or not; if the focus module (106) receives the light reflected from the sample (300), moving the lens module (104) to the sample (300) in a second set step size, less than the first set step size, along the optical axis and calculating a first light intensity parameter according to the light intensity of the light received by the focus module (106), and determining whether the first light intensity parameter is less than a first set light intensity threshold or not; and if the first light intensity parameter is less than the first set light intensity threshold, storing the current position of the lens module (104) as a storage position. The imaging method is convenient to operate and can reduce cost.

Description

Imaging method, device and system

Technical field

The present invention relates to the field of optical detection, and more particularly to an imaging method, apparatus and system.

Background technique

Sequencing, ie 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. From a functional control perspective, 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.

Currently, 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.

Summary of the invention

The 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 light a shaft 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 optical axis Moving to the first set position; moving the lens module from the first set position by the first set step along the optical axis 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 step smaller than the first set step Longly moving the sample along the optical axis, and calculating a first light intensity parameter according to the light intensity of the light received by the focusing module, determining whether the first light intensity parameter is smaller than the first setting a light intensity threshold; wherein the first light intensity parameter is less than the first Setting the intensity threshold, saving the current position of the lens module as the location.

In the above imaging method, by comparing the first light intensity parameter and the first set light intensity threshold, the interference of the non-sample strong reflected light signal on focusing/focusing can be eliminated; the detection and judgment based on the change of the optical signal can be quickly determined. Determining a specific location facilitates a faster and more accurate finding of the plane in which the target object is clearly imaged, ie a clear plane/clear surface. This imaging method is particularly suitable for devices that contain precise optical systems that are difficult to find clear planes, such as optical inspection equipment with high magnification lenses. This is easy to operate and reduces costs.

An optical detection system according to an embodiment of the present invention 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, causing the lens module to follow the sample in the optical axis at a second set step smaller than the first set step Moving, and calculating a first light intensity parameter according to the light intensity of the light received by the focusing module, determining whether the first light intensity parameter is smaller than a first set light intensity threshold; Saving the lens module when the strong parameter is less than the first set light intensity threshold The current location is the save location.

The optical detection system can eliminate the interference of the non-sample strong reflected light signal on focusing/focusing by comparing the first light intensity parameter with the first set light intensity threshold; detecting and judging based on the change of the optical signal can be fast Judging to determine a specific location facilitates further rapid and accurate finding of the plane in which the target object is clearly imaged, ie, a clear plane/clear surface. 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. This is easy to operate and reduces costs.

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.

Additional aspects and advantages of the embodiments of the invention will be set forth in part in

DRAWINGS

The above and/or additional aspects and advantages of the embodiments of the present invention will become apparent and readily understood from

1 is a schematic flow chart of an image forming method according to an embodiment of the present invention.

2 is a schematic view showing the positional relationship between a lens module and a sample according to an embodiment of the present invention.

3 is a partial structural schematic view of an optical detecting system according to an embodiment of the present invention.

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.

Figure 6 is a block diagram of an optical detection system in accordance with an embodiment of the present invention.

detailed description

The embodiments of the present invention are described in detail below, and the examples of the embodiments are illustrated in the drawings, wherein the same or similar reference numerals indicate the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the drawings are intended to be illustrative of the invention and are not to be construed as limiting.

In the description of the present invention, it is to be understood that the terms "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. Thus, features defining "first" or "second" may include one or more of the described features either explicitly or implicitly. In the description of the present invention, the meaning of "a plurality" is two or more unless specifically and specifically defined otherwise.

In the description of the present invention, it should be noted that, unless otherwise explicitly defined and defined, "connected" 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. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood on a case-by-case basis.

The following disclosure provides many different embodiments or examples for implementing different structures of the present invention. In order to simplify the disclosure of the present invention, the components and settings of specific examples are described below. In addition, the present invention may be repeated with reference to the numerals and/or reference numerals in the various examples, which are for the purpose of simplification and clarity, and do not indicate the relationship between the various embodiments and/or settings discussed.

In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", " After, "Left", "Right", "Vertical", "Horizontal", "Top", "Bottom", "Inside", "Outside", "Clockwise", "Counterclockwise", "Axial", The orientation or positional relationship of the "radial", "circumferential" and the like is based on the orientation or positional relationship shown in the drawings, and is merely for convenience of description of the present invention and simplified description, and does not indicate or imply the indicated device or component. It must be constructed and operated in a particular orientation, and is not to be construed as limiting the invention.

In the description of the present invention, 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. The so-called "sequence determination reaction" is the same as the sequencing reaction.

1 to 3, 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, and 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 toward the sample along the optical axis OP by a second set step smaller than the first set step, and is calculated according to the light intensity of the light received by the focus module 106. The first light intensity parameter determines whether the first light intensity parameter is smaller than the first set light intensity threshold; when the first light intensity parameter is less than the first set light intensity threshold, S15, saves the current position of the lens module 104 as a save position.

In the above imaging method, by comparing the first light intensity parameter and the first set light intensity threshold, the focus/focus interference of the non-sample strong reflected light signal, such as the oil surface/air reflected light signal of the objective lens 110, may be excluded; The detection and judgment based on the change of the optical signal can quickly determine the specific position, which is convenient for finding the plane of the target object clearly and quickly, that is, the clear plane/clear surface. This imaging method is particularly suitable for devices that contain precise optical systems that are difficult to find clear planes, such as optical inspection equipment with high magnification lenses. This is easy to operate and reduces costs.

Specifically, referring to FIG. 2, in the embodiment of the present invention, 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. Above the 200. 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.

In the embodiment of the present invention, since 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 subsequent acquisition of the clear image by the imaging device 102. In the embodiment of the present invention, the sample 302 to be tested is a solution, the front panel 202 of the carrying device 200 is glass, and 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. Therefore, in the embodiment of the present invention, by comparing the first light intensity parameter and the first set light intensity threshold, the strongly reflected light signal at the position of the non-media interface 204 can be excluded, such as the oil signal reflected by the oil level/air of the objective lens 110. Interference with focus/focus.

In one example, the front panel 202 of the sample 302 to be tested has a thickness of 0.175 mm.

In some embodiments, 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. In some embodiments, 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.

In some embodiments, referring to FIG. 3, 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.

In the embodiment of the present invention, 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. In addition, the microscope 107 further includes a barrel lens 111 between the objective lens 110 and the camera 108.

In some embodiments, 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.

In some embodiments, 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.

In addition, 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. In some examples, the imaging device 102 drives the objective lens 110 to move using a drive such as a stepper motor or a voice coil motor.

In some embodiments, when the coordinate system is established, as shown in FIG. 2, 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.

In one example, imaging device 102 includes a total internal reflection fluorescence microscope, objective lens 110 is 60x magnification, and the first set step size S1 = 0.01 mm. Thus, 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. In one example, the second set step size S3 = 0.002 mm. It is to be understood that, in other examples, other values may be used for the first set step size and the second set step size, which are not specifically limited herein.

When the focus module 106 does not receive the light reflected by the sample 300, the lens module 104 is caused to continue moving toward the sample 300 along the optical axis OP at the first set step.

When the first light intensity parameter is greater than or equal to the first set light intensity threshold, the lens module 104 is continuously moved along the optical axis OP by the second set step.

In some embodiments, the optical detection system can be applied to a sequencing system, or the sequencing system includes an optical detection system.

In some embodiments, 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. In this way, 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.

In some embodiments, such as in a total internal reflection imaging system, in order to quickly find the media interface, 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. For example, on a 60-fold total internal reflection imaging device with an objective lens, the range of movement of the lens module 104 can be set to 200 μm ± 10 μm or [190 μm, 250 μm] according to optical path characteristics and experience.

In some embodiments, 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. In one example, the second set position is set to be the lowest position of the upper surface 205 of the front panel of the reaction device 200 to the next depth of field, and the movement range of the lens module 104 is set to 250 μm. The location is determined. In the example of the present invention, 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.

Specifically, in the embodiment of the present invention, the movement range is an interval on the negative axis of the Z-axis. In one example, the first set position is nearlimit, the second set position is farlimit, and 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. In FIG. 2, 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.

It should be noted that in other embodiments, 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.

In some embodiments, 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.

Specifically, in the embodiment of the present invention, the light source 116 can be an infrared light source 116, and the light sensor 118 can be a photo diode. Thus, the cost is low and the detection accuracy is high. 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. In the embodiment of the present invention, when the sample 300 includes the carrier device 200 and the sample to be tested 302, the light reflected by the sample 300 is light reflected by the lower surface of the front panel of the carrier device 200.

Whether the infrared light reflected by the sample 300 enters the objective lens 110 and is received by the light sensor 118 depends mainly on the distance of the objective lens 110 from the sample 300. Therefore, when it is determined that the focus module 106 receives the infrared light reflected by the sample 300, it can be determined that the distance between the objective lens 110 and the sample 300 is moderate, and can be used for imaging of the imaging device 102. In one example, the above distance is 20-40 um.

At this time, 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.

In some embodiments, referring to FIG. 4, when the focus module 106 receives the light reflected by the sample 300, the method further includes the steps of:

S16, moving the lens module 104 to the sample 300 along the optical axis OP in a third setting step that is smaller than the first set step and greater than the second set step, and according to the light received by the focusing module 106. Calculating the second light intensity parameter by the light intensity, determining whether the second light intensity parameter is greater than the second set light intensity threshold;

When the second light intensity parameter is greater than the second set light intensity threshold, step S14 is performed.

In this way, by comparing the second light intensity parameter and the second set light intensity threshold, interference of focusing/focusing of the light signal which is very weak compared with the reflected light of the sample (such as the medium interface) can be excluded.

When the second light intensity parameter is not greater than the second set light intensity threshold, the lens module 104 is caused to continue moving toward the sample 300 along the optical axis OP in the third set step.

In one example, 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.

In some embodiments, the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the second light intensity parameter being the light of the light received by the two light sensors 118. Strong average. As such, the second 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 interference of the weak light signal with respect to focus/focus is more accurately excluded.

Specifically, the second light intensity parameter may be set to SUM, that is, SUM=(PD1+PD2)/2, and PD1 and PD2 respectively represent the light intensity of the light received by the two light sensors 118. In one example, the second set light intensity threshold nSum=40.

In some embodiments, the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the light intensity of the light received by the two light sensors 118 having a first difference The first light intensity parameter is a difference between the first difference value and the set compensation value. As such, the first light intensity parameter is calculated by the light intensity of the light received by the two light sensors 118 such that the interference of the highly reflected light signal with respect to focus/focus is more accurate.

Specifically, the difference (ie, the first light intensity parameter) may be set to err, and the offset value is set to offset, that is, err=(PD1-PD2)-offset. In an ideal state, the first difference can be zero. In one example, the first set light intensity threshold nErr=10, offset=30.

In some embodiments, referring to FIG. 5, when the first light intensity parameter is less than the first set light intensity threshold, the imaging method further includes the following steps: S17, making the lens module 104 smaller than the second set step The fourth set step moves along the optical axis OP and performs image acquisition on the sample 300 by the imaging device 102, and determines whether the sharpness value of the image collected by the imaging device 102 reaches a set threshold; the sharpness value of the image When the set threshold is reached, S18, the current position of the lens module 104 is saved to replace the previous save position. As such, the imaging device can clearly image the sample.

In the present embodiment, the sharpness value of the image can be used as an evaluation value of the 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.

When the sharpness value of the image reaches the set threshold, the current position of the lens module 104 is saved to replace the previous save position, so that the imaging device 102 can output a clear image when the sequence measurement reaction is taken.

In addition, the current position of the lens module 104 is saved to replace the previous save position, so that the save position is updated, and when the imaging device 104 is imaged, the sample 300 is image-collected with the saved position after the lens module 104 is updated.

It should be noted that the embodiment shown in FIG. 5 includes steps S16, S17, and S18. It can be understood that in other embodiments, step S16 can be omitted.

In some embodiments, when the lens module 104 is moved by the fourth 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 fourth 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 that is less than the fourth 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 fourth 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.

Specifically, the fourth set step size can be used as the coarse adjustment step length Z1, the fifth set step size can be used as the fine adjustment step size Z2, and 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.

Taking the current position of the lens module 104 as the starting point T, the coarse adjustment range Z3 is the adjustment range, that is, the adjustment range on the Z axis is (T, T+Z3). First, 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. The first sharpness value R1 of the image captured by the imaging device 102 and the second sharpness value R2 of the image captured by the imaging device 102 when the lens module 104 is in the previous position. R0 represents the set difference.

When 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.

When 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.

When 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.

When R2>R1 and R2-R1<R0, it means that the sharpness value of the image has crossed the set threshold and is closer to the set threshold, so that the lens module 104 has the step size Z2 in the opposite direction to the first direction. Move in the two directions and approach the set threshold with a smaller step size.

In some embodiments, when the lens module 104 is moved, 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 set difference can also be adjusted according to the distance from the peak of the sharpness value.

In one example, T=0, Z1=100, Z2=40, Z3=2100, and the adjustment range is (0, 2100). It should be noted that the above values are used for the metric value used when moving the lens module 104 during the image capturing process by the imaging device 102, and the metric value is related to the light intensity.

In some embodiments, the imaging 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; controlling the lens mode when the sample 300 is moved by the carrier The motion of group 104 is maintained to maintain the 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.

In particular, 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 200 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.

In some embodiments, 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.

Specifically, in the example of the present invention, the third set position may be nPos, the coordinate position corresponding to nPos is on the negative axis of the Z axis, and the coordinate position corresponding to nPos is greater than the coordinate position corresponding to the second set position farlimit. When the coordinate position corresponding to the current position of the lens module 104 is smaller than the coordinate position corresponding to the third set position, it is determined that the current position of the lens module 104 exceeds the third set position.

When it is first determined that the current position of the lens module 104 exceeds the third set position, refocusing is performed to adjust the position of the lens module 104 to attempt successful tracking. In 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.

In some embodiments, when the current position of the lens module 104 does not exceed the third set position, the relative position is determined to be unchanged. In some embodiments, 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.

Referring to FIG. 6, an optical detection system 100 according to an embodiment of the present invention 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 moves the sample module 300 along the optical axis OP to the sample 300 at a second set step smaller than the first set step, and receives the light according to the focus module 106. Calculating the first light intensity parameter to determine whether the first light intensity parameter is smaller than the first set light intensity threshold; when the first light intensity parameter is less than the first set light intensity threshold, saving the current state of the lens module 104 The location is the save location.

It should be noted that the explanation and description of the technical features and beneficial effects of the imaging method in any of the above embodiments and embodiments are also applicable to the optical detection system 100 of the present embodiment. To avoid redundancy, the details are not detailed here. Expand.

In some embodiments, 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.

In some embodiments, 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.

Specifically, the control device 101 can control the light source 116 to emit light and control the light sensor 118 to receive light.

In some embodiments, when the focus module 106 receives light reflected by the sample 300, the control device 101 is configured to:

The lens module 104 is moved toward the sample 300 along the optical axis OP by a third set step that is smaller than the first set step and larger than the second set step, and is based on the light intensity of the light received by the focus module 106. Calculating a second light intensity parameter, determining whether the second light intensity parameter is greater than a second set light intensity threshold;

When the second light intensity parameter is greater than the second set light intensity threshold, the lens module 104 is moved toward the sample 300 along the optical axis OP by a second set step smaller than the first set step, and Calculating a first light intensity parameter according to the light intensity of the light received by the focus module 106, and determining whether the first light intensity parameter is smaller than a first set light intensity threshold.

In some embodiments, the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the second light intensity parameter being the light of the light received by the two light sensors 118. Strong average.

In some embodiments, the focus module 106 includes two light sensors 118 for receiving light reflected by the sample 300, the light intensity of the light received by the two light sensors 118 having a first difference The first light intensity parameter is a difference between the first difference value and the set compensation value.

In some embodiments, when the first light intensity parameter is less than the first set light intensity threshold, the control device 101 is configured to:

The lens module 104 is moved along the optical axis OP by a fourth set step smaller than the second set step and the image is collected by the imaging device 102, and the sharpness value of the image collected by the imaging device 102 is determined. Whether the set threshold is reached;

When the sharpness value of the image reaches the set threshold, the current position of the lens module 104 is saved to replace the previous save position.

In some embodiments, when the lens module 104 is moved by the fourth 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 second sharpness value of the image corresponding to the previous position of the group 104; the sharpness difference between the first sharpness value and the second sharpness value is greater than the first sharpness value is greater than the second sharpness value When the difference is fixed, the lens module 104 is caused to continue moving along the optical axis OP to the sample 300 in a fourth set step; the first sharpness value is greater than the second sharpness value and the first sharpness value and the second sharpness are When the sharpness difference between the values 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 fourth 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 value and the sharpness difference between the second sharpness value and the first sharpness value is greater than the set difference value , the lens module 104 is moved away from the sample 300 along the optical axis OP by a fourth set step; the second sharpness value is greater than the first sharpness value and the second sharpness value is When the sharpness difference between the first sharpness values 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 image collected by the imaging device 102. The sharpness value reaches the set threshold.

In some embodiments, 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 the second set position; and the current position of the lens module 104 exceeds the second set position. At the same time, stop moving the lens module 104 or focus.

Specifically, when the control device 101 performs focusing, the focusing step in the method of the above embodiment can be performed.

In some embodiments, the 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.

In some embodiments, 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. When 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.

Referring to FIG. 6, 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. For storing data, 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.

In the description of the present specification, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiment", "example", "specific example", or "some examples", etc. The specific features, structures, materials or characteristics described in the embodiments or examples are included in at least one embodiment or example of the invention. In the present specification, the schematic representation of the above terms does not necessarily mean the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in a suitable manner in any one or more embodiments or examples.

The logic and/or steps represented in the flowcharts or otherwise described herein, for example, may be considered as an ordered list of executable instructions for implementing logical functions, and may be embodied in any computer readable storage medium. For use by an instruction execution system, apparatus, or device (eg, a computer-based system, a system including a processor, or other system that can fetch instructions and execute instructions from an instruction execution system, apparatus, or device), or in conjunction with such instruction execution systems, Used for devices or equipment. For the purposes of this specification, 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. . More specific examples (non-exhaustive list) of 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). In addition, 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.

In addition, 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.

Although the embodiments of the present invention have been shown and described, it is understood that the above-described embodiments are illustrative and are not to be construed as limiting the scope of the invention. The embodiments are subject to changes, modifications, substitutions and variations.

Claims

An imaging method, characterized in that the method is used in an optical detection system, the optical detection system comprising an imaging device and a stage, the imaging device comprising a lens module and a focusing module, the lens module comprising light A shaft for carrying a sample, the method comprising the following focusing steps:

Using the focusing module to emit light onto the sample placed on the stage;

Moving the lens module along the optical axis to a 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 be along the optical axis in a second setting step smaller than the first set step Moving the sample, and calculating a first light intensity parameter according to the light intensity of the light received by the focusing module, determining whether the first light intensity parameter is less than a first set light intensity threshold;

When the first light intensity parameter is smaller than the first set light intensity threshold, the current position of the lens module is saved as a save position.
The method of claim 1 wherein said focusing module comprises a light source for emitting said light onto said sample, and said light sensor for receiving reflection from said sample The light.
The method of claim 1 wherein when said focusing module receives said light reflected by said sample, said method further comprising the steps of:

Moving the lens module along the optical axis at a third setting step that is smaller than the first set step and greater than the second set step, and according to the focus mode Calculating a second light intensity parameter by the light intensity of the received light, and determining whether the second light intensity parameter is greater than a second set light intensity threshold;

When the second light intensity parameter is greater than the second set light intensity threshold, performing the second set step of the lens module to be smaller than the first set step along the optical axis And moving the sample to calculate a first light intensity parameter according to the light intensity of the light received by the focusing module, and determining whether the first light intensity parameter is smaller than a first set light intensity threshold.
The method of claim 3 wherein said focus module comprises two light sensors for receiving said light reflected by said sample, said second light intensity parameter An average of the light intensities of the light received by the two photosensors.
The method of claim 1 wherein said focus module comprises two light sensors for receiving said light reflected by said sample, said two light sensors receiving The light intensity of the received light has a first difference, and the first light intensity parameter is a difference between the first difference and the set compensation value.
The method according to any one of claims 1 to 5, wherein when the first light intensity parameter is smaller than the first set light intensity threshold, the method further comprises the following steps:

Moving the lens module along the optical axis in a fourth set step size smaller than the second set step and performing image acquisition on the sample by using the imaging device, and determining the imaging device Whether the sharpness value of the collected image reaches a set threshold;

When the sharpness value of the image reaches the set threshold, the current position of the lens module is saved to replace the save position.
The method according to claim 6, wherein when the lens module is moved in the fourth set step, the first of the patterns corresponding to the current position of the lens module is determined. Whether the sharpness value is greater than a second sharpness value of the image corresponding to a previous position of the lens module;

And 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 a set difference value, The module continues to move the sample along the optical axis in the fourth set step;

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 less than the set difference value, The lens module continues to move the sample along the optical axis at a fifth set step size smaller than the fourth set step to enable the sharpness value of the image collected by the imaging device to reach Setting the 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 value, Moving the lens module away from the sample along the optical axis in the fourth set step;

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 less than the set difference value, The lens module moves away from the sample along the optical axis in the fifth set step to cause the sharpness value of the image collected by the imaging device to reach the set threshold.
The method according to any one of claims 1 to 7, wherein when the lens module is moved, it is determined whether the current position of the lens module exceeds a second set position;

When the current position of the lens module exceeds the second set position, the moving of the lens module or the performing the focusing step is stopped.
The method of any of claims 1-8, wherein the method further comprises the following scoring step:

Determining a relative position of the lens module and the sample when the lens module is in the saving position;

When the sample is moved by the stage, the movement of the lens module is controlled to keep the relative position unchanged.
The method according to claim 9, wherein when the sample is moved by the stage, it is determined whether the current position of the lens module exceeds a third set position;

When the current position of the lens module exceeds the third set position, the sample is moved by the stage and the focusing step is performed;

When the number of movements of the sample reaches a set number of times and the current position of the lens module still exceeds the third set position, it is determined that the focus recovery has failed.
An optical detection system, comprising: a control device, an imaging device and a stage, the imaging device comprising a lens module and a focusing module, the lens module comprising an optical axis, the carrier for carrying a sample The control device is used to:

Using the focusing module to emit light onto the sample placed on the stage;

Moving the lens module along the optical axis to a 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 be along the optical axis in a second setting step smaller than the first set step Moving the sample, and calculating a first light intensity parameter according to the light intensity of the light received by the focusing module, determining whether the first light intensity parameter is less than a first set light intensity threshold;

When the first light intensity parameter is smaller than the first set light intensity threshold, the current position of the lens module is saved as a save position.
The system of claim 11 wherein said focus module comprises a light source for emitting said light onto said sample, and said light sensor for receiving reflection from said sample The light.
The system of claim 11 wherein said control means is operative to: when said focus module receives said light reflected by said sample:

Moving the lens module along the optical axis at a third setting step that is smaller than the first set step and greater than the second set step, and according to the focus mode Calculating a second light intensity parameter by the light intensity of the received light, and determining whether the second light intensity parameter is greater than a second set light intensity threshold;

When the second light intensity parameter is greater than the second set light intensity threshold, causing the lens module to be along the optical axis at a second set step size smaller than the first set step The sample is moved, and the first light intensity parameter is calculated according to the light intensity of the light received by the focus module, and it is determined whether the first light intensity parameter is smaller than the first set light intensity threshold.
The system of claim 13 wherein said focus module comprises two light sensors for receiving said light reflected by said sample, said second light intensity parameter An average of the light intensities of the light received by the two photosensors.
The system of claim 11 wherein said focus module comprises two light sensors for receiving said light reflected by said sample, said two light sensors receiving The light intensity of the received light has a first difference, and the first light intensity parameter is a difference between the first difference and the set compensation value.
The system of claim 11 wherein said control means is operative to: when said first intensity parameter is less than said first set light intensity threshold:

Moving the lens module along the optical axis in a fourth set step size smaller than the second set step and performing image acquisition on the sample by using the imaging device, and determining the imaging device Whether the sharpness value of the collected image reaches a set threshold;

When the sharpness value of the image reaches the set threshold, the current position of the lens module is saved to replace the save position.
The system of claim 16 wherein said control means is operative to determine a current position of said lens module when said lens module is moved in said fourth set step Whether the first sharpness value of the pattern is greater than a second sharpness value of the image corresponding to a previous position of the lens module;

And 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 a set difference value, The module continues to move the sample along the optical axis in the fourth set step;

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 less than the set difference value, The lens module continues to move the sample along the optical axis at a fifth set step size smaller than the fourth set step to enable the sharpness value of the image collected by the imaging device to reach Setting the 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 value, Moving the lens module away from the sample along the optical axis in the fourth set step;

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 less than the set difference value, The lens module moves away from the sample along the optical axis in the fifth set step to cause the sharpness value of the image collected by the imaging device to reach the set threshold.
The system according to any one of claims 11-17, wherein the control device is configured to determine whether the current position of the lens module exceeds a second set position when the lens module is moved;

When the current position of the lens module exceeds the second set position, the lens module is stopped from moving or focusing.
A system according to any one of claims 11 to 18, wherein said control means is for:

Determining a relative position of the lens module and the sample when the lens module is in the saving position;

When the sample is moved by the stage, the movement of the lens module is controlled to keep the relative position unchanged.
The system according to claim 19, wherein said control means is configured to determine whether a current position of said lens module exceeds a third set position when said sample is moved by said stage;

When the current position of the lens module exceeds the third set position, the sample is moved by the stage and focused;

When the number of movements of the sample reaches a set number of times and the current position of the lens module still exceeds the third set position, it is determined that the focus recovery has failed.
A control device for controlling imaging for an optical detection system, the optical detection system comprising an imaging device and a focusing module, wherein the control device comprises:

a storage device for storing data, the data comprising a computer executable program;

A processor for executing the computer executable program, the executing the computer executable program comprising performing the method of any of claims 1-10.