A method and system for dynamic through focus scanning imaging

Abstract

The application relates to a dynamic over-focus scanning imaging method, and belongs to the technical field of optical microscopic imaging. The method solves the problem that when the existing over-focus scanning imaging method is used to scan along the Z axis, the X axis / Y axis is static, and the detection efficiency is low. The method comprises the following steps: setting an over-focus scanning range; continuously and periodically moving a sample along the Z axis in the over-focus scanning range while continuously moving the sample along the X axis / Y axis relative to a plane array camera; the moving speed of the Z axis, the X axis / Y axis satisfies that when the Z axis moves a distance S Z , the X axis / Y axis moves one pixel, and every time the Z axis moves a distance S Z , the plane array camera takes a photo to record the object surface information of the position; until the X axis / Y axis of the sample is scanned, one scanning is completed; the sample is moved along the Y axis / X axis to the next camera field of view range, and scanning along the X axis / Y axis is continued until the whole sample is scanned; and scanning imaging data obtained is processed to obtain final scanning imaging data along the Z axis at each X axis / Y axis position.

Description

Technical Field

[0001] This invention relates to the field of optical microscopy imaging technology, and in particular to a method and system for dynamic overfocus scanning imaging. Background Technology

[0002] In recent years, optical microscopy has been developing rapidly. In order to meet the needs of scanning and detecting three-dimensional objects, optical microscopy has received widespread attention and in-depth research.

[0003] Through-focus scanning optical microscopy (TSOM) is an image-based three-dimensional measurement method in the field of optical microscopy that scans near the focal point along the optical axis (Z-axis). This method possesses optical super-resolution capabilities, capturing a single focused image and scanning along the optical axis to obtain an image sequence consisting of a focused image of the imaging target and a series of defocused images, forming a data cube. Comparison and processing of these images improve imaging resolution and accuracy. TSOM can achieve super-resolution using ordinary optical microscopes, and its simple structure, ease of operation, and low cost have led to its widespread application in three-dimensional surface imaging and inspection.

[0004] Traditional TSOM, such as Figure 2 As shown, scanning is performed along the Z-axis. After scanning one field of view, the field of view is moved to a neighboring area by X-axis / Y-axis translation, and then the neighboring area is scanned along the Z-axis. Finally, the data is summarized and stitched together to obtain the scanning data information of the entire sample. When the sample size is large, this independent working mode of X-axis / Y-axis scanning and Z-axis scanning significantly reduces imaging efficiency and makes it impossible to achieve rapid online detection of large samples. Summary of the Invention

[0005] Based on the above analysis, the present invention aims to provide a method and system for dynamic overfocus scanning imaging, in order to solve the problems of low efficiency and poor real-time performance of existing overfocus scanning imaging methods.

[0006] This invention provides a method for dynamic overfocus scanning imaging, the method comprising the following steps:

[0007] Set the defocus range of the scanning imaging according to the sample size and measurement accuracy;

[0008] The sample is continuously and periodically moved along the Z-axis within the defocus range, while simultaneously moving continuously relative to the area array camera along the X / Y axes; the magnitudes of the moving speeds along the Z-axis and X / Y axes satisfy the condition that the moving distance S along the Z-axis... Z When the X / Y axis moves exactly one pixel, and the Z axis moves a distance S... ZThe area array camera takes a picture to record the object surface information at the position; until the X axis / Y axis of the sample is scanned, a scan is completed;

[0009] The sample is moved to the next camera field of view along the Y axis / X axis, and the scan is continued along the X axis / Y axis until the entire sample is scanned;

[0010] The obtained scan imaging data is processed to obtain the final scan imaging data along the Z axis at each X axis / Y axis position.

[0011] Further, the continuous periodic movement of the sample along the Z axis within the defocus range while the sample is continuously moved relative to the area array camera along the X axis / Y axis comprises:

[0012] In one scan, the sample is reciprocally moved along the Z axis, and the sample is continuously moved relative to the area array camera along the X axis / Y axis.

[0013] Further, the reciprocally moving of the sample along the Z axis comprises:

[0014] The sample is moved along the Z axis by a distance S Z The sample is moved relative to the area array camera along the X axis / Y axis by a distance of one pixel, and each time the Z axis is moved by a distance S Z The area array camera takes a picture to record the object surface information at the position, and obtains a scan image at the corresponding Z axis position and X axis / Y axis position; the sample is moved from top to bottom or from bottom to top along the Z axis, so that the area array camera scans from the top to the bottom or from the bottom to the top of the sample defocus range for a half cycle, and the area array camera takes 2N-1 times of pictures in one cycle; the sample is continuously moved for one cycle for one reciprocating motion.

[0015] Further, the distance S Z of the sample along the Z axis during the scan imaging is set according to the size of the sample and the measurement accuracy.

[0016] Further, the area array camera is M columns, and has P1 to P M pixels, wherein M≥2N-1.

[0017] Further, the processing of the obtained scan imaging data to obtain the final scan imaging data along the Z axis at each X axis / Y axis position comprises:

[0018] A data table is established with each X axis / Y axis position as a row and each pixel as a column, the obtained scan imaging data is stored in the corresponding position of the information table; according to the information table, imaging data containing each Z axis position Z1 to Z N is extracted at each X axis / Y axis position, redundant data is filtered out, and the extracted data is sorted from Z1 to Z NThe final scanning imaging data along the Z axis at each X axis / Y axis position can be obtained by sorting the imaging data in the order of the Z axis.

[0019] Further, when there are multiple imaging data at the same Z axis position at an X axis / Y axis position, the imaging data at the first same Z axis position is taken as the final value of the Z axis position.

[0020] Further, the sample includes a three-dimensional surface object and an optically transparent three-dimensional object.

[0021] Further, the dynamic through-focus scanning imaging system comprises the following modules:

[0022] A moving device for moving the sample;

[0023] A surface array camera for scanning imaging the sample;

[0024] A data acquisition device for acquiring the scanning imaging data of the surface array camera and transmitting the scanning imaging data to a data processing module;

[0025] A data processing module for processing the scanning imaging data to obtain final scanning imaging data.

[0026] Further, the system further comprises a controller for controlling the moving device to move the sample along the X axis / Y axis continuously relative to the surface array camera while the sample is continuously and periodically moved along the Z axis within the defocus range; when the Z axis moves a distance S Z , the X axis / Y axis moves exactly one pixel; after the surface array camera completes a scanning, the sample is moved to the next camera field of view range along the Y axis / X axis and continues to scan along the X axis / Y axis until the entire sample is scanned.

[0027] Compared with the prior art, the present application can at least achieve one of the following beneficial effects:

[0028] 1. The present application changes the traditional through-focus scanning static scanning mode and is a dynamic through-focus scanning (Dynamic TSOM) method, the Z axis scanning and the X axis / Y axis scanning are performed simultaneously, which greatly shortens the scanning time, improves the speed and detection efficiency of online detection, and meets the TSOM detection requirements when the sample size is large.

[0029] 2. The dynamic through-focus scanning method of the present application can be applied to the scanning of various samples, including three-dimensional surface objects and optically transparent three-dimensional objects.

[0030] The technical solutions in the present application can be combined with each other to realize more preferred combination solutions. Other features and advantages of the present application will be described in the following description, and some advantages will become apparent from the description, or will be learned by practice of the present application. The objects and other advantages of the present application can be realized and obtained by the content particularly pointed out in the description and the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0031] The accompanying drawings are included to provide a further understanding of the present application, and are incorporated in and constitute a part of this application. The drawings illustrate embodiments of the present application and, together with the description, serve to explain the principles of the present application.

[0032] Figure 1 A flow chart of a dynamic overfocus scanning imaging method according to an embodiment of the present application.

[0033] Figure 2 A traditional overfocus scanning imaging method.

[0034] Figure 3 A working principle diagram of a dynamic overfocus scanning imaging method according to an embodiment of the present application.

[0035] Figure 4 A dynamic overfocus scanning imaging method according to an embodiment of the present application.

[0036] Figure 5 A system device diagram according to an embodiment of the present application. DETAILED DESCRIPTION

[0037] Preferred embodiments of the present application will be described in detail below with reference to the drawings, which form a part of this application. The drawings, together with the description, are used to explain the principles of the present application, and are not intended to limit the scope of the present application.

[0038] One specific embodiment of the present application discloses a dynamic overfocus scanning imaging method.

[0039] As shown in Figure 1 , the method comprises the following steps:

[0040] Step S1: setting an overfocus range of scanning imaging according to the size of a sample and the measurement accuracy.

[0041] Step S2: continuously and periodically moving the sample along the Z axis within the overfocus range while continuously moving the sample relative to the area array camera along the X axis / Y axis; the moving speed of the Z axis, the X axis / Y axis satisfies that when the Z axis moves a distance S Z , the X axis / Y axis moves exactly one pixel, and every time the Z axis moves a distance S Z , the area array camera takes a picture to record the object surface information at the position; until the X axis / Y axis of the sample is scanned, one scanning is completed.

[0042] Step S3: Move the sample along the Y-axis / X-axis to the next camera field of view, and continue scanning along the X-axis / Y-axis until the entire sample is scanned.

[0043] Step S4: Process the acquired scanning imaging data to obtain the final scanning imaging data along the Z-axis at each X-axis / Y-axis position.

[0044] In step S1, the defocus range refers to the range of images acquired by the area array camera at the out-of-focus position. In practice, the defocus range can be set according to the size of the sample and the measurement accuracy.

[0045] In step S2, the following steps are performed to obtain the scanning imaging data after each movement.

[0046] Step S2.1: Set the distance S that the sample moves along the Z-axis during scanning imaging based on the sample size and measurement accuracy. Z .

[0047] Step S2.2: In a single scan, the sample moves continuously and periodically along the Z-axis within the defocus range while simultaneously moving continuously relative to the area array camera along the X / Y axes; the magnitudes of the moving speeds along the Z-axis and X / Y axes satisfy the condition that when the Z-axis moving distance S... Z At that time, the X-axis / Y-axis moves exactly one pixel; such as Figure 4 As shown.

[0048] Specifically, the Z-axis includes N positions, and the sample moves a distance S along the Z-axis. Z This means obtaining a Z-axis position, such as Z1; the sample moves a distance S along the Z-axis. Z Simultaneously, the sample moves one pixel relative to the area scan camera along the X / Y axis, thus obtaining an X / Y axis position, such as X1 / Y1; the sample moves along the Z axis from top to bottom or from bottom to top, that is, from Z1 to Z. N Or from Z N Moving to Z1, the area array camera scans from the top to the bottom or from the bottom to the top of the sample's out-of-focus area, which is half a cycle; the sample moves continuously for one cycle, which is one reciprocating motion.

[0049] Step S2.3: Whenever the Z-axis moves a distance S Z The area array camera takes pictures and records the object surface information at that location, obtaining scan images at the corresponding Z-axis and X / Y-axis positions; within one cycle, the area array camera takes 2N-1 pictures, that is, for every movement distance S... Z It moves one pixel along the X / Y axis and takes one picture.

[0050] Specifically, the area scan camera is arranged in M ​​columns, with P1 to P2. M Each pixel.

[0051] Step S2.4: Record the scanning imaging data after each movement to obtain the initial scanning imaging data along the Z-axis at each X-axis / Y-axis position.

[0052] Specifically, in order to ensure that the scanning imaging data along the Z-axis at each X / Y axis position can contain a complete cycle, M needs to satisfy M≥2N-1.

[0053] Let M = 2N-1, such as Figure 3 and Figure 4 As shown, at each X-axis / Y-axis position, the sample was photographed M times, resulting in 2N-1 corresponding Z-axis positions. Therefore, at each X-axis / Y-axis position, N Z-axis positions will be traversed, meaning there are N overfocus scanning imaging data at different Z-axis positions.

[0054] Specifically, the sample moves one pixel relative to the area scan camera along the X / Y axis, including the following two methods:

[0055] The area scan camera remains stationary, while the sample moves one pixel along the X / Y axis; the sample remains stationary along the X / Y axis, while the area scan camera moves one pixel along the X / Y axis. In step S3, after the sample has moved several pixels relative to the area scan camera along the X / Y axis, the area scan camera has completed scanning the sample along the X / Y axis. Then, the sample is moved along the Y / X axis within the camera's field of view, and scanning continues along the X / Y axis until the entire sample has been scanned.

[0056] Specifically, if the area scan camera has completed scanning the sample's X-axis, the sample is moved along the Y-axis by one camera field of view, and then the scanning continues; if the area scan camera has completed scanning the sample's Y-axis, the sample is moved along the X-axis by one camera field of view, and then the scanning continues.

[0057] In step S4, the following steps are performed to obtain the final scan imaging data.

[0058] Step S4.1: Create a data table with each X-axis / Y-axis position as the row and each pixel position as the column, and store the acquired scanning imaging data in the corresponding position of the information table.

[0059] A specific example of an information table is as follows:

[0060]

[0061] Step S4.2: Based on the information table, extract the Z-axis positions Z1 to Z2 at each X-axis / Y-axis position. N The imaging data is filtered to remove redundant data.

[0062] Specifically, when there are multiple imaging data points at the same Z-axis position at a given X-axis / Y-axis position, the imaging data point at the first Z-axis position is taken as the final value of that Z-axis position, while the imaging data points at other Z-axis positions are deleted.

[0063] For example, if there are two imaging data points for the X2 position, the imaging data of the first Z2 position is taken as the imaging data of the Z2 position, and the imaging data of the second Z2 position is deleted.

[0064] Step S4.3: Process the extracted data according to Z1 to Z... N By sorting the data in order, the final scan imaging data along the Z-axis at each X / Y-axis position can be obtained.

[0065] As shown in the information table above, for example at position X2, it corresponds to P1 to P M The corresponding Z-axis position is Z2Z N Z N-1 Z1Z2, the extracted data order is Z2Z N Z1 is not the sequence from Z1 to Z. N order ...

[0067] Therefore, the extracted data needs to be sorted from Z1 to Z. N Sort them in the order they appear.

[0068] Another specific embodiment of the present invention discloses a system for dynamic overfocus scanning imaging. For example... Figure 5 As shown, the system includes the following modules:

[0069] A moving device used to move the sample;

[0070] Area array camera, used for scanning and imaging samples;

[0071] A data acquisition device is used to acquire scanning imaging data from an area array camera and transmit the scanning imaging data to a data processing module.

[0072] The data processing module is used to process the scanned imaging data to obtain the final scanned imaging data.

[0073] Specifically, the system also includes a controller for controlling the moving device, causing the moving device to continuously and periodically move the sample along the Z-axis within the defocus range while simultaneously moving the sample continuously relative to the area array camera along the X / Y axes; when the Z-axis movement distance S... ZThe X / Y axis moves exactly one pixel; after the area scan camera completes one scan, the sample is moved along the Y / X axis to the next camera's field of view, and the scanning continues along the X / Y axis until the entire sample is scanned.

[0074] Compared with existing technologies, this invention provides a dynamic overfocus scanning imaging method. First, it changes the traditional static scanning mode of overfocus scanning to a dynamic overfocus scanning (Dynamic TSOM) method. By delaying the time, it can perform X / Y axis scanning simultaneously with Z-axis scanning, which will greatly shorten the scanning time, improve the speed and efficiency of online detection, and meet the TSOM detection requirements when the sample size is large. Second, the dynamic overfocus scanning method can be applied to the scanning of various samples, including three-dimensional surface objects and optically transparent three-dimensional objects.

[0075] Furthermore, the definition of the above method is not limited to the various specific forms, structures, shapes or methods mentioned in the embodiments. For example, the shooting interval can be any reasonable value; M and N can be any values ​​that satisfy the condition M≥2N-1.

[0076] Those skilled in the art will understand that all or part of the processes of the methods described in the above embodiments can be implemented by a computer program instructing related hardware, and the program can be stored in a computer-readable storage medium. The computer-readable storage medium may be a disk, optical disk, read-only memory, or random access memory, etc.

[0077] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for dynamic overfocus scanning imaging, characterized in that, The method includes the following steps: Set the defocus range of the scanning imaging according to the sample size and measurement accuracy; The sample is continuously and periodically moved along the Z-axis within the defocus range while simultaneously moving continuously relative to the area array camera along the X-axis or Y-axis, including: in one scan, the sample reciprocates along the Z-axis while simultaneously moving continuously relative to the area array camera along the X-axis or Y-axis. The sample reciprocating motion along the Z-axis includes: the sample moving from top to bottom or from bottom to top along the Z-axis, such that the area array camera scans from the top to the bottom or from the bottom to the top of the sample's defocus range as half a cycle, and the area array camera takes 2N-1 pictures in one cycle; the sample continuously moving for one cycle is one reciprocating motion. The magnitude of the movement speed along the Z-axis, X-axis, or Y-axis satisfies the condition that the distance moved along the Z-axis... When the X-axis or Y-axis moves exactly one pixel, whenever the Z-axis moves a certain distance... The temporal array camera captures and records the surface information of the object at the current position; a scan is completed when the X-axis or Y-axis of the sample has been scanned. Move the sample along the Y-axis or X-axis to the next camera field of view, and continue scanning along the X-axis or Y-axis until the entire sample has been scanned; The acquired scanning imaging data is processed to obtain the final scanning imaging data along the Z-axis at each X-axis or Y-axis position. This includes: establishing a data table with each X-axis or Y-axis position as a row and each pixel as a column; storing the acquired scanning imaging data in the corresponding positions of the information table; and extracting the Z-axis position data from each X-axis or Y-axis position based on the information table. to The imaging data is filtered to remove redundant data, and the extracted data is processed according to... to By sorting the data in order, the final scan imaging data along the Z-axis can be obtained at each X-axis or Y-axis position. When there are multiple imaging data at the same Z-axis position at an X-axis or Y-axis position, the imaging data at the first Z-axis position is taken as the final value of that Z-axis position.

2. The dynamic overfocus scanning imaging method according to claim 1, characterized in that, The cyclic reciprocating motion of the sample along the Z-axis also includes: Sample movement distance along the Z-axis At any given time, the sample moves one pixel relative to the area array camera along the X-axis or Y-axis, and each time it moves a distance along the Z-axis... The time-area array camera captures and records the object surface information at the current position, obtaining scanned images at the corresponding Z-axis position and X-axis or Y-axis position.

3. The dynamic overfocus scanning imaging method according to claim 2, characterized in that, Set the distance the sample moves along the Z-axis during scanning imaging based on the sample size and measurement accuracy. .

4. The dynamic overfocus scanning imaging method according to claim 2, characterized in that, The area array camera is in M ​​columns, with to There are 2N-1 pixels, where M ≥ 2N-1.

5. The dynamic overfocus scanning imaging method according to claim 1, characterized in that, The samples include three-dimensional surface objects and optically transparent three-dimensional objects.

6. A dynamic overfocus scanning imaging system, characterized in that, The method for implementing the dynamic overfocus scanning imaging method of claim 1 includes the following modules: A moving device used to move the sample; Area array camera, used for scanning and imaging samples; A data acquisition device is used to acquire scanning imaging data from an area array camera and transmit the scanning imaging data to a data processing module. The data processing module is used to process the scanned imaging data to obtain the final scanned imaging data; The system also includes a controller for controlling the moving device, causing the moving device to continuously and periodically move the sample along the Z-axis within the defocus range while simultaneously moving the sample continuously relative to the area array camera along the X-axis or Y-axis; when the Z-axis movement distance... The X-axis or Y-axis moves exactly one pixel; after the area array camera completes one scan, the sample is moved along the Y-axis or X-axis to the next camera's field of view, and the scanning continues along the X-axis or Y-axis until the entire sample is scanned.

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