CN110162351B - Software system for three-dimensional scanning optical microscopic imaging - Google Patents
Software system for three-dimensional scanning optical microscopic imaging Download PDFInfo
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Abstract
The invention provides a software system for three-dimensional scanning optical microscopic imaging, which mainly comprises a data acquisition module, a display module, a control module and a data storage module; the software system is developed on the basis of MFC by utilizing VC + + in a VS2013 compiling environment; the data acquisition module comprises a photon acquisition part and a nanometer displacement part; the nanometer displacement part comprises a two-dimensional nanometer displacement part and a three-dimensional nanometer displacement part; the software system focuses on the compiling of imaging software of a common Laser Scanning Confocal Microscope (LSCM), a fluorescence lifetime imaging system (FLIM), a stimulated emission depletion microscope (STED) and other systems, the parameter setting mode is simple, the interface is simple and attractive, the humanized design meets the experimental requirements and can contain all required function realization, the software system is very suitable for low development cost and rapid development conditions, the practical requirements under the scientific and technological rapid development background are met, and the software system is very suitable for the construction and related application of an optical microscope system.
Description
Technical Field
The invention relates to the technical field of microscopic imaging software, in particular to a software system for three-dimensional scanning optical microscopic imaging.
Background
The optical microscope is used for high-precision imaging of subcellular structures, and has the advantages of no damage, real time and the like, so that the optical microscope is always a main research means of life science, cell biology and nano material science. However, the commercial system of Laser Scanning Confocal Microscope (LSCM) commonly used in laboratories is expensive, which limits its widespread use in life sciences. Meanwhile, for scientific research purposes based on different needs, for example, a super-resolution optical imaging system for realizing ultrahigh spatial resolution exceeding the optical diffraction limit often needs to change an optical path or modify the system, and the development of such experiments is not facilitated by using a highly centralized commercialized system. In addition, people pursue high imaging resolution, and meanwhile, the intelligent control of the imaging system realizes high requirements on the parameter setting flexibility, imaging quality, system stability and the like of the imaging software system. Therefore, the self-constructed scanning imaging microscope and the corresponding software system development can adapt to different requirements, the cost can be saved, and the wide application of high-resolution optical microscopic imaging is promoted.
Disclosure of Invention
The invention aims to provide a software system for three-dimensional scanning optical microscopic Imaging, which can be used for constructing a laser confocal microscope system (LSCM), can also be directly used for a Fluorescence Lifetime Imaging (FLIM) and other super-resolution Imaging technologies, such as the system development of a Stimulated Emission Depletion microscope (STED), promotes the application of the optical microscopic Imaging technology in the fields of life science, nano science and the like, promotes the development of the related fields, and has important practical application value.
In order to achieve the purpose, the invention provides the following technical scheme: a software system for three-dimensional scanning optical microscopic imaging mainly comprises a data acquisition module, a display module, a control module and a data storage module;
the software system is developed on the basis of MFC by utilizing VC + + in a VS2013 compiling environment;
the data acquisition module comprises a photon acquisition part and a nanometer displacement part, realizes the accurate synchronous control of photon acquisition and nanometer displacement, scans each pixel by adopting a mode of gradually moving and gradually recording the photon number of a single pixel, and realizes the time consistency of the photon number recording of the single pixel and the pixel position moving; the nanometer displacement part comprises a two-dimensional nanometer displacement part and a three-dimensional nanometer displacement part; the two-dimensional nanometer displacement part moves in three modes of transverse XY scanning, longitudinal XZ scanning and YZ scanning; the three-dimensional nanometer displacement part controls the nanometer translation stage to carry out two-dimensional XY scanning at a randomly set Z-axis position;
the display module comprises an image display part, a photon number real-time display part and a parameter real-time display part;
the control module mainly comprises a parameter setting part and a start-stop control part;
the data storage module is divided into image storage and data storage.
Furthermore, the photon collecting part is composed of a single photon detector and a photon collecting card, photon information of each imaging pixel point is collected, photon counting is achieved, the nanometer displacement part controls the nanometer translation platform through a computer, and accurate movement between the imaging sample and the scanning lens in a three-dimensional space is achieved.
Furthermore, the software system realizes the time synchronization control of the photon acquisition card and the nano translation stage in a software code mode;
the time synchronization control is realized by calling corresponding interface functions of a photon acquisition card and a nano translation stage, the acquisition interface function of the photon acquisition card is called to acquire photon numerical values of single time and single pixel, and a moving interface function of the nano translation stage is called to realize the movement of the position of a pixel point; the photon acquisition card acquires the photon numerical value once when the nano translation stage moves one position, and the photon acquisition card starts and stops once and has high time synchronization.
Furthermore, the start-stop control part calls a hardware drive function to realize control through a button message response function.
Further, the software system implementation method includes: signal transmission and data acquisition, pixel gray level calibration, multithreading programming, parameter setting and real-time display and data storage;
the signal transmission is realized by adopting an SMA communication cable, and the signal is a pulse signal of a single photon detector;
photon number information of each pixel acquired by the data acquisition module is stored in a self-defined three-dimensional array in a one-to-one correspondence manner according to a set scanning time interval and the positions of X, Y and Z three-dimensional spaces of corresponding nano translation stages, and the photon number information is used for image display and data storage of OpenGl;
the image display is that OpenGl calibrates the gray value of each imaging pixel according to photon information of a data acquisition module to reconstruct a two-dimensional image; the gray value is calibrated according to 256 gray levels according to the magnitude of the photon value, and the corresponding relation can be obtained by using the following formula:
wherein S represents the gray level of the pixel point, C c Representing the number of photons of the pixel currently to be calibrated, C mi Representing the minimum photon number, C, in a three-dimensional array ma Representing the maximum photon value in the three-dimensional array;
the data acquisition module and the display module are programmed in a multi-thread mode, and the multi-thread mode comprises two threads, a parameter display thread and a scanning thread;
the parameter setting part comprises parameter setting of a photon acquisition card and relevant parameter setting of a nano translation stage, and parameter setting of the photon acquisition card is carried out in an EditControl control by establishing a modeless dialog box; the relevant parameters of the nano translation stage are set by an edit box control; the parameter setting part can set the acquisition time interval of a single pixel of the photon acquisition card, various parameters of the photon acquisition card, the single movement step length of the nano translation stage and the scanning range;
the real-time display comprises real-time display of photon numbers and real-time display of parameters, the real-time display of the photon numbers is realized by a TeeChart control, and the real-time display of the parameters is realized by an edit box and a progress bar; the real-time parameter display is realized by independently starting a thread;
the data storage comprises image storage and original data storage, wherein the image storage adopts an interface screen capture mode, is stored in a BITMAP format and is stored in a current directory; and storing the original data in a TXT format to a current working directory, wherein the original data is photon information data stored in the three-dimensional array and is the result of single imaging scanning, and the last scanning result is covered after the next scanning is started.
Further, the software system operation process can be divided into: initializing a software system, acquiring and processing information, scanning and moving a sampling point position, running multithreading, performing internal memory processing on photon information, displaying an image and displaying the number of photons in real time;
the software system initialization is realized by adopting MFC basic controls, and firstly, various parameters of the photon acquisition card are set: scanning time intervals, TAC gain, TAC range, CFD threshold value, CFD upper and lower limits, and setting the moving range and moving step length of the nano translation stage;
the information acquisition processing is that a single photon detector converts a photon pulse signal into an electric pulse signal, and the electric pulse signal is input into a time-dependent single photon acquisition card through an SMA cable to realize the photon information acquisition of each pixel; the photon information acquisition comprises the steps of storing data into a three-dimensional array according to a set scanning time interval and corresponding X, Y and Z positions;
the scanning movement of the sampling point position is realized by a computer and a nano translation stage, namely the nano translation stage receives a computer command to perform accurate movement, so that the scanning of a three-dimensional space is realized;
the operation of the multiple threads is as follows: starting the parameter display thread immediately after the system initialization is finished, and ending when the software interface is closed; the scanning thread is started after the two-dimensional or three-dimensional scanning is started, and the one-time two-dimensional or three-dimensional scanning of the graph is finished;
the memory processing of the photon information is that an FIFO buffer caches in a mode of accessing immediately;
the image display is realized in a point tracing mode by adopting OpenGl, the pixel value of each point is calibrated according to the photon number of each pixel point of a data acquisition module, and a two-dimensional image is reconstructed by combining the three-dimensional array; the three-dimensional array is a system memory space defined by a programmer, the software system enables the three-dimensional coordinates of the nano translation stage to correspond to the positions of the three-dimensional array one by one, corresponding photon numerical values are stored in the corresponding positions, and the photon numerical values are converted into gray values of pixel points at the corresponding positions when an image is reconstructed;
the photon number is displayed in real time by drawing FastLine by a TeeChart control; the device comprises a TeeCart control, a TeeCart control and a photon curve, wherein the TeeCart control is used for controlling the TeeCart control to be used, the photon number acquired by a photon acquisition part at a single time is taken as a longitudinal axis, the time interval of current acquisition is taken as a transverse axis, the photon curve is drawn in real time, the real-time photon curve reflects the current scanning position in real time, the photon number acquired in a unit time interval is acquired, and the TeeCart control needs to be provided with a TeeCart 8.Ocx Active component before being used.
Further, the software system operation process can be described as:
(1) Connecting the nano translation stage and the photon acquisition card;
(2) Clicking a setting button to open a modeless dialog box to set various parameters of the photon acquisition card;
(3) Setting parameters of acquisition time interval, scanning range and scanning step length in a control interface, and setting a scanning initial position according to requirements;
(4) The system initialization is finished when the setting is finished;
(5) Starting scanning, namely starting a ComBox control for controlling scanning, starting a scanning thread by selecting a scanning mode, and executing a corresponding imaging scanning process;
(6) In the scanning process, a user can see a real-time photon number curve, and when a single imaging scanning process is finished, a scanning image can be presented in an OpenGl area of a control interface;
(7) The user can click the save button respectively to store the scanned image and the original data.
Compared with the prior art, the invention has the beneficial effects that:
the software system focuses on the compiling of imaging software of a common Laser Scanning Confocal Microscope (LSCM), a fluorescence lifetime imaging system (FLIM), a stimulated emission depletion microscope (STED) and other systems, the parameter setting mode is simple, the interface is simple and attractive, the humanized design meets the experimental requirements and can contain all required function realization, the software system is very suitable for low development cost and rapid development conditions, the practical requirements under the scientific and technological rapid development background are met, and the software system is very suitable for the construction and related application of an optical microscope system.
Drawings
FIG. 1 is a system interface diagram of the present invention;
FIG. 2 is a schematic diagram of two-dimensional scanning imaging according to the present invention;
FIG. 3 is a two-dimensional scanning imaging exemplary effect diagram of the present invention;
FIG. 4 is a schematic diagram of three-dimensional scanning imaging according to the present invention;
FIG. 5 is a partial view of a real-time photon counting display interface in accordance with the present invention;
FIG. 6 is a flow chart of the system of the present invention;
FIG. 7 is a flow chart of the system initialization of the present invention;
FIG. 8 is a flowchart of a scanning imaging process of the present invention;
FIG. 9 is a system memory processing profile of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to the accompanying drawings and embodiments. The embodiments described herein are only for explaining the technical solution of the present invention and are not limited to the present invention.
The software system provided by the invention is developed on the basis of MFC by utilizing VC + + in VS2013 compiling environment. The system adopts a multithreading programming mode to realize simultaneous data display and three-dimensional space scanning. The software system realizes the position movement of the three-dimensional space between the fluorescent sample and the objective lens by controlling the nano translation stage, thereby realizing the three-dimensional point scanning of the imaging system. Meanwhile, the software system controls the photon acquisition card to receive and count photon pulse information of the single photon detector. The software system can set the scanning step length and the scanning range of the single movement of the nano translation stage and the single acquisition time interval of the photon acquisition card in a control interface. The software system realizes two-dimensional and three-dimensional image reconstruction by using OpenGl and realizes real-time photon number display by using a TeeChart control. The software system stores the image in bitmap format and stores the data in txt format.
According to the software implementation method provided by the invention, firstly, parameters such as scanning step length, scanning range, single pixel acquisition time interval and the like are set on a control interface shown in figure 1, then, a scanning mode is set, and a 'start scanning' button is clicked to start software imaging. Two-dimensional scanning imaging is schematically shown in fig. 2, and the system adopts a point scanning mode. Taking transverse (XY) two-dimensional scanning as an example, when a Y axis is at an initial position, the computer controls the nano translation stage to move along the X axis, and when a pixel is moved, the photon acquisition card counts an acquisition time interval, records the sum of photon numbers in the scanning time interval set by software as a pixel intensity value, and stores each pixel intensity value to a corresponding position of a self-defined three-dimensional array of the memory buffer area. And scanning the nano translation stage along the X-axis direction according to the set scanning range and the single pixel acquisition time interval, and continuously increasing the step length along the X-axis until the preset scanning range in the X direction is reached. Then, the scanning along the X axis is repeated by moving one step along the Y axis. When the Y axis is increased by one scanning step, the X axis finishes one line of scanning to form an S-shaped scanning path. Until both the X-direction and the Y-direction reach the maximum range at the same time, i.e., the two-dimensional scan ends. Figure 3 gives an example of two-dimensional imaging with 40 nm fluorescent beads and 90 nm gold particles. Fig. 4 shows a schematic diagram of three-dimensional scanning imaging, namely, three-dimensional scanning, namely, on the basis of two-dimensional scanning, the movement of the Z axis is increased. On the basis of two-dimensional scanning, when each two-dimensional image is scanned, the corresponding computer controls the Z-axis direction to move upwards or downwards by a step length until the set Z-axis scanning range is completed, and then the two-dimensional images of different Z axes are layered and superposed to reconstruct a three-dimensional image.
The three-dimensional array is a system memory space defined by a programmer, the software system enables the three-dimensional coordinates of the nano translation stage to correspond to the positions of the three-dimensional array one by one, corresponding photon numerical values are stored in the corresponding positions, and the photon numerical values are converted into gray values of pixel points in the corresponding positions during image reconstruction. The three-dimensional array is used for reconstructing two-dimensional images at the end of scanning and storing data of single scanning.
The typical scan range for a transverse (XY) two-dimensional scan is 10um × 10um; a typical value of the scanning step is 0.1um; the acquisition time interval is typically 0.01s. It should be understood, however, that the scan range, scan step size, and acquisition time interval are not limited to the typical values described above, depending on the fluorescent sample and imaging requirements.
According to the software implementation method provided by the invention, openGl is utilized to realize reconstruction and display of two-dimensional and three-dimensional images. After the position information of each pixel point and the gray value corresponding to the position information are extracted from the three-dimensional matrix, each pixel in the three-dimensional array corresponds to one pixel on the software interface in a point drawing mode, the gray value also corresponds to the pixel, and each pixel point is completely drawn to present a plane image.
And the TeeChart control is utilized to realize real-time display of photon numbers. The invention selects a Fastline drawing mode of a TeeChart control to realize the real-time display function of the photon number. As shown in fig. 5, each scanning time interval scans a point, and displays the photon count of the pixel point in real time. The points are connected into a curve to obtain a real-time photon number curve along with the accumulation of the scanning time.
The gray value is calibrated according to 256 gray levels according to the magnitude of the photon value, and the corresponding relation can be obtained by using the following formula:
wherein S represents the gray level of the pixel point, C c Representing the number of photons of the pixel currently to be calibrated, C mi Representing the minimum photon number, C, in a three-dimensional array ma Representing the maximum photon value in the three-dimensional array.
Before OpenGl is used, an OpenGl common library (glut32. Lib, glut32.Dll, glut.h and the like) needs to be installed in a VS2013 environment. Before the TeeChart control is used, the TeeChart8.Ocx Active control needs to be reinstalled in a Windows operating system and successfully registered.
According to the software implementation method provided by the invention, the data storage of the software is divided into image storage and data storage. The image storage adopts a screenshot mode, a button 'image storage' designed by software is clicked, and the image is stored in a BITMP format. The data is stored in the TXT format under the current working directory. The original data is photon information data stored in the customized three-dimensional array.
The interface and the specific implementation flow of the software system are specifically described below with reference to the accompanying drawings:
FIG. 1 is a schematic diagram of a software system. The upper left corner is a ListBox control used for displaying the current state and parameter setting condition of the system; the middle black frame is an OpenGl interface and is used for displaying a two-dimensional image; on the right is a TeeChart control for displaying photon number changes in real time. The real-time display of parameters such as counting rate of the photon acquisition card is arranged at the lower left part; the middle part can display information such as the maximum photon number of each scanning; the lower part of the TeeChart control is a setting area of parameters such as a system scanning time interval, a scanning step length and the like; the bottom is a control area for controlling the starting and stopping of the system and storing data.
As shown in fig. 6, the software system is initialized first after being started. And after the system initialization is finished, setting system parameters, and after the setting is finished, judging whether the parameter setting is finished or not, so as to prevent omission. And then the system receives a scanning starting command through the interactive interface to perform scanning imaging. The system finishes one two-dimensional scanning and stops scanning. And storing the image data and the original data through a human-computer interaction interface.
As shown in fig. 7, the system initialization mainly includes the connection of the nano translation stage and the start-up of the photon acquisition card; setting system parameters such as the moving range and the moving step length of the nano translation stage; and (4) setting parameters such as scanning time interval, TAC range, gain, CFD upper and lower limits and the like of the photon acquisition card.
As shown in fig. 8, the scanning imaging process mainly solves the precise synchronous control of photon collection and nanometer displacement. Firstly, the position of the nano translation stage is reset to zero, the photon collecting card is started to count the total number of photons within a certain time interval, whether the counting time reaches a preset value or not is judged, and if the counting time reaches the preset time value, the nano translation stage moves by one scanning step length. And meanwhile, judging whether the nano translation stage moves to a preset maximum range or not, if not, moving the nano translation stage to a new position, and counting and storing the total number of photons in the same time interval by the photon acquisition card. And scanning each pixel in a mode of gradually moving and gradually recording the photon number of a single pixel, and repeating the whole process until the nano translation stage moves to a preset maximum range. After the scanning is finished, the software system starts OpenGl to reconstruct a two-dimensional image and displays the two-dimensional image on an interface.
As shown in fig. 9, the memory of the software system is obtained by FIFO buffer, and is transferred to the internal self-defined three-dimensional array of the buffer area by a way of storing and fetching, and then the system extracts the data in the three-dimensional array to reconstruct the two-dimensional image and store the data.
The foregoing description merely represents preferred embodiments of the present invention, which are described in some detail and detail, and should not be construed as limiting the scope of the present invention. It should be noted that various changes, modifications and substitutions may be made by those skilled in the art without departing from the spirit of the invention, and all are intended to be included within the scope of the invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (6)
1. A software system for three-dimensional scanning optical microscopy imaging is characterized in that: the software system mainly comprises a data acquisition module, a display module, a control module and a data storage module;
the software system is developed on the basis of MFC by utilizing VC + + in a VS2013 compiling environment;
the data acquisition module comprises a photon acquisition part and a nanometer displacement part, realizes the accurate synchronous control of photon acquisition and nanometer displacement, scans each pixel by adopting a mode of gradually moving and gradually recording the photon number of a single pixel, and realizes the time consistency of the photon number recording of the single pixel and the pixel position moving; the nanometer displacement part comprises a two-dimensional nanometer displacement part and a three-dimensional nanometer displacement part; the two-dimensional nanometer displacement part moves in three modes of transverse XY scanning, longitudinal XZ scanning and YZ scanning; the three-dimensional nanometer displacement part controls the nanometer translation stage to carry out two-dimensional XY scanning at a randomly set Z-axis position;
the display module comprises an image display part, a photon number real-time display part and a parameter real-time display part;
the control module mainly comprises a parameter setting part and a start-stop control part;
the data storage module is divided into image storage and data storage;
the software system implementation method comprises the following steps: signal transmission and data acquisition, pixel gray level calibration, multithreading programming, parameter setting and real-time display and data storage;
the signal transmission is realized by adopting an SMA communication cable, and the signal is a pulse signal of a single-photon detector;
photon number information of each pixel acquired by the data acquisition module is stored in a self-defined three-dimensional array in a one-to-one correspondence manner according to a set scanning time interval and the positions of the corresponding X, Y and Z three-dimensional spaces of the nano translation stage, and the photon number information is used for image display and data storage of OpenGl;
the image display is that OpenGl calibrates the gray value of each imaging pixel according to photon information of a data acquisition module to reconstruct a two-dimensional image; the gray value is calibrated according to 256 gray levels according to the magnitude of the photon value, and the corresponding relation can be obtained by using the following formula:
wherein S represents the gray level of the pixel point, C c Representing the number of photons of the pixel currently to be calibrated, C mi Representing the minimum photon number, C, in a three-dimensional array ma Representing the maximum photon value in the three-dimensional array;
the data acquisition module and the display module are programmed in a multi-thread mode, and the multi-thread mode comprises two threads, a parameter display thread and a scanning thread;
the parameter setting part comprises parameter setting of a photon acquisition card and relevant parameter setting of a nano translation stage, and parameter setting of the photon acquisition card is carried out in an EditControl control by establishing a modeless dialog box; the relevant parameters of the nano translation stage are set by an edit box control; the parameter setting part can set the acquisition time interval of a single pixel of the photon acquisition card, various parameters of the photon acquisition card, the single movement step length of the nano translation stage and the scanning range;
the real-time display comprises real-time photon number display and real-time parameter display, wherein the real-time photon number display is realized by a TeeChart control, and the real-time parameter display is realized by an edit box and a progress bar; the real-time parameter display is realized by independently starting threads;
the data storage comprises image storage and original data storage, wherein the image storage adopts an interface screen capture mode, is stored in a BITMAP format and is stored in a current directory; the original data is stored in a TXT format and is stored in a current working directory, the original data is photon information data stored in the three-dimensional array and is the result of single imaging scanning, and the last scanning result is covered after the next scanning is started.
2. The software system for three-dimensional scanning optical microscopy imaging as defined in claim 1, wherein: the photon collecting part is composed of a single photon detector and a photon collecting card, photon information of each imaging pixel point is collected, photon counting is achieved, the nanometer displacement part controls the nanometer translation stage through a computer, and accurate movement between the imaging sample and the scanning lens in a three-dimensional space is achieved.
3. The software system for three-dimensional scanning optical microscopy imaging according to claim 1, characterized in that: the software system realizes the time synchronous control of the photon acquisition card and the nano translation stage in a software code mode;
the time synchronization control is realized by calling corresponding interface functions of a photon acquisition card and a nano translation stage, the acquisition interface function of the photon acquisition card is called to acquire single-time and single-pixel photon numerical values, and the moving interface function of the nano translation stage is called to realize the movement of the position of a pixel point; the photon collecting card can obtain the photon value once when the nanometer translation stage moves one position, and the photon collecting card is started and stopped once and highly time-synchronized.
4. The software system for three-dimensional scanning optical microscopy imaging as defined in claim 1, wherein: the start-stop control part calls a hardware drive function through a button message response function to realize control.
5. The software system for three-dimensional scanning optical microscopy imaging according to claim 1, characterized in that: the software system operation process can be divided into: initializing a software system, acquiring and processing information, scanning and moving a sampling point position, running multithreading, processing a photon information memory, displaying an image and displaying photon number in real time;
the software system initialization is realized by adopting MFC basic controls, and firstly, the parameters of the photon acquisition card are set as follows: scanning time intervals, TAC gains, TAC ranges, CFD thresholds and CFD upper and lower limits of a photon acquisition card, and then setting the moving range and the moving step length of a nano translation stage;
the information acquisition processing is that a single photon detector converts a photon pulse signal into an electric pulse signal, and the electric pulse signal is input into a time-dependent single photon acquisition card through an SMA cable to realize the photon information acquisition of each pixel; the photon information acquisition comprises the steps of storing data into a three-dimensional array according to a set scanning time interval and corresponding X, Y and Z positions;
the scanning movement of the sampling point position is realized by a computer and a nano translation stage, namely the nano translation stage receives a computer command to perform accurate movement, so that the scanning of a three-dimensional space is realized;
the operation of the multiple threads is as follows: starting the parameter display thread immediately after the system initialization is finished, and ending when the software interface is closed; the scanning thread is started after the two-dimensional or three-dimensional scanning is started, and the one-time two-dimensional or three-dimensional scanning of the graph is finished;
the memory processing of the photon information is that an FIFO buffer caches in a mode of accessing and fetching instantly;
the image display is realized in a point tracing mode by adopting OpenGl, the pixel value of each point is calibrated according to the photon number of each pixel point of a data acquisition module, and a two-dimensional image is reconstructed by combining the three-dimensional array; the three-dimensional array is a system memory space defined by a programmer, the software system enables the three-dimensional coordinates of the nano translation stage to correspond to the positions of the three-dimensional array one by one, corresponding photon numerical values are stored in the corresponding positions, and the photon numerical values are converted into gray values of pixel points at the corresponding positions when an image is reconstructed;
the photon number is displayed in real time by drawing FastLine through a TeeChart control; the device comprises a TeeChart control and a TeeChart control, wherein the TeeChart control is used for drawing a photon curve in real time by taking the photon number acquired by a photon acquisition part at a single time as a longitudinal axis and the time interval of current acquisition as a horizontal axis, the real-time photon curve reflects the current scanning position in real time and the photon number acquired in a unit time interval, and a TeeChart8.Ocx Active component is required to be installed before the TeeChart control is used.
6. The software system for three-dimensional scanning optical microscopy imaging according to claim 1, characterized in that: the software system operation process can be described as follows:
(1) Connecting the nano translation stage and the photon acquisition card;
(2) Clicking a setting button to open a modeless dialog box to set various parameters of the photon acquisition card;
(3) Setting parameters of acquisition time interval, scanning range and scanning step length in a control interface, and setting a scanning initial position according to requirements;
(4) The system initialization is finished when the setting is finished;
(5) Starting scanning, namely starting a ComBox control for controlling scanning, starting a scanning thread by selecting a scanning mode, and executing a corresponding imaging scanning process;
(6) In the scanning process, a user can see a real-time photon number curve, and when a single imaging scanning process is finished, a scanning image can be presented in an OpenGl area of a control interface;
(7) The user can click the save button respectively to store the scanned image and the original data.
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| CN113091606B (en) * | 2021-03-21 | 2022-11-11 | 西北工业大学 | Cross-scale micro-nano structure laser manufacturing detection system and control method |
| CN113485700B (en) * | 2021-07-07 | 2022-06-28 | 华中科技大学鄂州工业技术研究院 | Design method and device of optical-mechanical-electrical modular control software based on Qt |
| CN114935815A (en) * | 2022-05-26 | 2022-08-23 | 南昌大学 | Microscopic imaging system and method based on scanning galvanometer and nano translation stage |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1453734A (en) * | 2002-04-23 | 2003-11-05 | 北京清大德人显微数字技术有限公司 | Computerized laser confocal scanning microscope system |
| JP2006084960A (en) * | 2004-09-17 | 2006-03-30 | Yokogawa Electric Corp | Three-dimensional confocal microscopic system |
| CN103487419A (en) * | 2013-09-22 | 2014-01-01 | 江苏美伦影像系统有限公司 | Confocal laser scanning micro-imaging system |
-
2019
- 2019-03-12 CN CN201910182445.9A patent/CN110162351B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1453734A (en) * | 2002-04-23 | 2003-11-05 | 北京清大德人显微数字技术有限公司 | Computerized laser confocal scanning microscope system |
| JP2006084960A (en) * | 2004-09-17 | 2006-03-30 | Yokogawa Electric Corp | Three-dimensional confocal microscopic system |
| CN103487419A (en) * | 2013-09-22 | 2014-01-01 | 江苏美伦影像系统有限公司 | Confocal laser scanning micro-imaging system |
Non-Patent Citations (1)
| Title |
|---|
| 《MFC中利用TeeChart控件完成三维图形绘制》;神马观止;《https://blog.csdn.net/iloveyoumj/article/details/18460925》;20140118;第1-5页 * |
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