CN109444093B - Synchronous imaging detection method and system - Google Patents
Synchronous imaging detection method and system Download PDFInfo
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- CN109444093B CN109444093B CN201811087103.0A CN201811087103A CN109444093B CN 109444093 B CN109444093 B CN 109444093B CN 201811087103 A CN201811087103 A CN 201811087103A CN 109444093 B CN109444093 B CN 109444093B
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
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- G01N21/84—Systems specially adapted for particular applications
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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Abstract
The embodiment of the invention provides a synchronous imaging detection method and a synchronous imaging detection system, wherein the method comprises the steps of obtaining a plasma state optical signal generated by each three-dimensional measuring point on a blade after being excited by a first laser beam and a fluorescence signal generated after being excited by a second laser beam; obtaining an element distribution map of the nutrient elements to be measured in the leaves based on the content of the nutrient elements to be measured in all the three-dimensional measuring points; and obtaining a molecular spectrogram of the molecules to be detected in the blade based on the content of the molecules to be detected in all the three-dimensional measuring points. The synchronous imaging detection method and the synchronous imaging detection system provided by the embodiment of the invention adopt the first laser beam and the second laser beam to sequentially focus all three-dimensional measurement points of the blade to obtain the fluorescence signal generated by the molecule to be detected at each three-dimensional measurement point and the plasma state optical signal generated by the nutrient element to be detected, and obtain the spatial distribution of the molecule to be detected and the nutrient element to be detected in the blade through the analysis of all the signals.
Description
Technical Field
The embodiment of the invention relates to the technical field of plant detection, in particular to a synchronous imaging detection method and system.
Background
The plant leaves are the most sensitive parts reflecting the physiological metabolism condition, nutrient condition and the adaptation degree to the growth environment of the plants, and the main nutrient content can indirectly reflect the fertility condition of the soil, so that the difference of various nutrients in different plant leaves is analyzed, and a theoretical basis can be provided for the accurate regulation and control of the nutrients. Meanwhile, the plant leaves are used as places for photosynthesis and main organs for substance production, are energy converters of primary producers in an ecological system, are vigorous in physiological activity, sensitive to environmental change and strong in plasticity, and the nutrient composition, transportation, transformation and accumulation of the plant leaves can reflect the physiological and ecological conditions of plants and represent the environmental conditions adapted to the plants. Therefore, the rapid and in-situ detection of the nutrient (particularly nitrogen) and chlorophyll content of the plant leaves is of great significance for understanding the absorption, transportation, transformation and accumulation of plant nutrient elements, the interaction between plants and the environment, accurate fertilization, pest and disease prevention and water and soil pollution prevention.
The existing detection methods for substances in plant tissue leaves comprise atomic absorption spectrometry, ICP emission spectrometry, an electrochemical method, laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS) and the like.
Among these methods, atomic absorption spectrometry, ICP emission spectrometry and electrochemical methods can only analyze one substance at a time, and require sampling and sample preparation, and the analysis process is complicated, costly and long in cycle. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS) are mainly used for detecting the element composition on the surface of a solid, and the element in the plant tissue is difficult to detect. Therefore, the existing leaf nutrient detection method cannot simultaneously detect the content contrast distribution condition of two substances in the leaf at corresponding positions in the distribution space.
Disclosure of Invention
Aiming at the defects in the prior art in the background art, the embodiment of the invention provides a synchronous imaging detection method and a synchronous imaging detection system.
In a first aspect, an embodiment of the present invention provides a synchronous imaging detection method, where the method includes:
acquiring a plasma state optical signal generated after each three-dimensional measuring point on the blade is excited by a first laser beam and a fluorescence signal generated after each three-dimensional measuring point is excited by a second laser beam;
obtaining the content of the nutrient elements to be measured in each three-dimensional measuring point on the blade based on the plasma state optical signal, and obtaining an element distribution map of the nutrient elements to be measured in the blade based on the content of the nutrient elements to be measured in all the three-dimensional measuring points; and obtaining the content of the molecules to be detected in each three-dimensional measuring point on the blade based on the fluorescence signals, and obtaining a molecular spectrogram of the molecules to be detected in the blade based on the content of the molecules to be detected in all the three-dimensional measuring points.
In a second aspect, an embodiment of the present invention provides a synchronous imaging detection system, where the system includes:
first formation of image detecting system, second formation of image detecting system and master control device, wherein:
the first imaging detection system is used for acquiring a plasma state optical signal generated by each three-dimensional measuring point on the blade after the first laser beam is excited, and acquiring a corresponding optical signal characteristic spectral line based on the plasma state optical signal;
the second imaging detection system is used for acquiring a fluorescence signal generated by each three-dimensional measuring point on the blade after being excited by the second laser beam, and acquiring a corresponding pixel point array database based on the fluorescence signal;
the main control device is used for obtaining the content of the nutrient elements to be detected in each three-dimensional measuring point of the blade based on the optical signal characteristic spectral line corresponding to each three-dimensional measuring point, obtaining an element distribution diagram of the nutrient elements to be detected in the blade based on the content of the nutrient elements to be detected in all the three-dimensional measuring points, obtaining the content of the molecules to be detected in each three-dimensional measuring point based on the pixel lattice column database corresponding to each three-dimensional measuring point, and obtaining a molecular spectrogram of the molecules to be detected in the blade based on the content of the molecules to be detected in all the three-dimensional measuring points.
The synchronous imaging detection method and the synchronous imaging detection system provided by the embodiment of the invention adopt the first laser beam and the second laser beam to focus all three-dimensional measurement points in a mode of sequentially focusing the same three-dimensional measurement point of the blade to obtain fluorescence generated after the molecules to be detected in each three-dimensional measurement point are excited and plasma state optical signals generated after the nutrient elements to be detected are excited, and obtain the content contrast distribution conditions of the molecules to be detected and the corresponding positions of the nutrient elements to be detected in the distribution space in the blade through the analysis of the fluorescence signals generated by the molecules to be detected in all the three-dimensional measurement points and the plasma state optical signals generated by the nutrient elements to be detected.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and those skilled in the art can also obtain other drawings according to the drawings without creative efforts.
FIG. 1 is a schematic flow chart of a synchronous imaging detection method according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of a synchronous imaging detection system according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of laser focusing of a synchronous imaging detection system according to an embodiment of the present invention;
fig. 4 is a schematic view of a workflow of a synchronous imaging detection system according to an embodiment of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments, but not all embodiments, of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The existing methods for detecting substances in plant leaf tissues comprise an atomic absorption spectrometry, an ICP emission spectrometry, an electrochemical method, a laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), a laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS) and the like, wherein in the methods, the atomic absorption spectrometry, the ICP emission spectrometry and the electrochemical method can only analyze one substance at a time, sampling and sample preparation are needed, the nutrient analysis process needs to be carried out in a laboratory, and the analysis process is complex, high in cost and long in period. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and laser microdissection inductively coupled plasma mass spectrometry (LMD-ICP-MS) are mainly used for detecting the element composition on the surface of a solid, and the element in the plant tissue is difficult to detect. The existing leaf nutrient detection method cannot simultaneously detect the content comparison distribution condition of two substances in the leaf at corresponding positions in a distribution space.
In order to solve the problem of simultaneously detecting the content contrast distribution condition of two substances in a distribution space, based on the LIBS (Laser-Induced Breakdown Spectroscopy) technology and the LIF (Laser-Induced Fluorescence) technology, the embodiment of the present invention provides a synchronous imaging detection method, and fig. 1 is a schematic flow chart of the synchronous imaging detection method provided by the embodiment of the present invention, as shown in fig. 1, the method includes:
step 11, obtaining the content of the nutrient elements to be detected in each three-dimensional measuring point on the blade based on the plasma state optical signal, and obtaining an element distribution map of the nutrient elements to be detected in the blade based on the content of the nutrient elements to be detected in all the three-dimensional measuring points; and obtaining the content of the molecules to be detected in each three-dimensional measuring point on the blade based on the fluorescence signals, and obtaining a molecular spectrogram of the molecules to be detected in the blade based on the content of the molecules to be detected in all the three-dimensional measuring points.
Specifically, the synchronous imaging detection method provided in the embodiment of the present invention is specifically to obtain a plasma state optical signal generated after excitation of a first laser beam by each three-dimensional measurement point on a blade and a fluorescence signal generated after excitation of a second laser beam, where the plasma state optical signal and the fluorescence signal corresponding to each three-dimensional measurement point are both obtained synchronously, that is, after obtaining the plasma state optical signal and the fluorescence signal corresponding to one three-dimensional measurement point, a plasma state optical signal and a fluorescence signal corresponding to the next three-dimensional measurement point are obtained, so as to obtain the plasma state optical signal and the fluorescence signal corresponding to each three-dimensional measurement point; analyzing the plasma state optical signal corresponding to each three-dimensional measuring point to obtain the content of the nutrient element to be measured in each three-dimensional measuring point, and obtaining a three-dimensional element distribution map capable of reflecting the spatial distribution of the nutrient element to be measured on the blade according to the content of the nutrient element to be measured in all the three-dimensional measuring points and the three-dimensional coordinates of each three-dimensional measuring point; the content of the molecules to be measured in each three-dimensional measuring point is obtained by analyzing the fluorescent signal corresponding to each three-dimensional measuring point, and a three-dimensional molecular spectrogram capable of reflecting the spatial distribution of the molecules to be measured on the blade is obtained according to the content of the molecules to be measured in all the three-dimensional measuring points and the three-dimensional coordinates of each three-dimensional measuring point.
Among them, the LIBS (Laser-Induced Breakdown Spectroscopy) technique is a brand new substance element analysis technique based on atomic emission Spectroscopy. The working principle is as follows: under the action of strong laser pulse, the matter on the surface of the sample is excited into plasma and is attenuated rapidly, photons with specific frequency are radiated in the attenuation process, characteristic spectral lines are generated, and the frequency and the intensity of the characteristic spectral lines contain the element type and content information of an analysis object. The LIF (Laser-Induced Fluorescence) technology belongs to the Laser spectrum molecular analysis technology, and adopts the principle that Laser irradiates the surface of an object to be detected, molecules of the object enter an excited state from a ground state after absorbing light energy, and are immediately de-excited to emit Fluorescence. It can be used not only for imaging but also for analysis of some elements. The method has the advantages of low operation cost, high measurement speed, high sensitivity, no or little sample pretreatment, no radiation and secondary pollution, capability of realizing multi-element measurement and the like, and has wide application prospect.
The synchronous imaging detection method provided by the embodiment of the invention adopts the mode that the first laser beam and the second laser beam sequentially focus the same three-dimensional measuring point of the blade to focus all the three-dimensional measuring points to obtain fluorescence generated after the molecules to be detected in all the three-dimensional measuring points are excited and plasma state optical signals generated after the nutrient elements to be detected are excited, and obtains the content contrast distribution conditions of the molecules to be detected and the nutrient elements to be detected at corresponding positions in the distribution space in the blade by analyzing the fluorescence signals generated by the molecules to be detected in all the three-dimensional measuring points and the plasma state optical signals generated by the nutrient elements to be detected.
On the basis of the foregoing embodiments, the acquiring a plasma state optical signal generated by each three-dimensional measurement point on a blade after being excited by a first laser beam and a fluorescence signal generated after being excited by a second laser beam in the synchronous imaging detection method provided by the embodiments of the present invention specifically includes:
the method comprises the steps that a first laser beam and a second laser beam are converged to a three-dimensional measuring point on a blade, and a plasma state optical signal generated after the first laser beam is excited and a fluorescence signal generated after the second laser beam is excited corresponding to the three-dimensional measuring point are obtained;
and sequentially focusing all three-dimensional measuring points on the blade through the first laser beam and the second laser beam. The specific method for acquiring the plasma state optical signal generated by each three-dimensional measuring point on the blade after being excited by the first laser beam and the fluorescence signal generated after being excited by the second laser beam by the synchronous imaging detection method provided by the embodiment of the invention is as follows: focusing all three-dimensional measuring points in a mode that a first laser beam and a second laser beam focus on the same three-dimensional measuring point of a blade to obtain a plasma state optical signal generated after the excitation of a nutrient element to be measured and a fluorescence signal generated after the excitation of a molecule to be measured in each three-dimensional measuring point, wherein the sequence of the first laser beam and the second laser beam focusing on the same three-dimensional measuring point of the blade can be selected according to specific needs, namely the first laser beam and the second laser beam can be focused simultaneously; or after the first laser beam is focused, focusing the second laser beam; for example, taking the spatial distribution of nitrogen elements and chlorophyll molecules as an example, the order of the same three-dimensional measurement point of the blade where the first laser beam and the second laser beam are focused may be: focusing a first three-dimensional measuring point of the blade by a second laser beam, exciting chlorophyll molecules in the three-dimensional measuring point, generating a fluorescent signal corresponding to the chlorophyll molecules, focusing the same three-dimensional measuring point of the blade, namely the first three-dimensional measuring point, by the first laser beam, exciting a nutrient element to be detected corresponding to the three-dimensional measuring point, generating a plasma state optical signal corresponding to the nutrient element to be detected, focusing a second three-dimensional measuring point of the blade by the second laser beam, re-exciting the chlorophyll molecules in the three-dimensional measuring point, generating a fluorescent signal corresponding to the chlorophyll molecules, focusing the second three-dimensional measuring point by the first laser beam, sequentially focusing the second three-dimensional measuring point by the second laser beam and the first laser beam, completing sequential excitation of the chlorophyll molecules and nitrogen elements in all the three-dimensional measuring points, and thus obtaining the fluorescent signal generated by the chlorophyll molecules in all the three-dimensional measuring points and the plasma state optical signal generated by the nitrogen element to be detected Number (n).
On the basis of the foregoing embodiments, the method for detecting synchronous imaging according to the embodiments of the present invention, which is based on plasma state optical signals, obtains the content of nutrient elements to be detected in each three-dimensional measurement point on a blade, and specifically includes:
and obtaining corresponding optical signal characteristic spectral lines based on plasma state optical signals generated after each three-dimensional measuring point is excited, and obtaining the content of the nutrient elements to be measured in each three-dimensional measuring point based on the optical signal characteristic spectral lines.
After a plasma state optical signal generated after the nutrient element to be detected is excited is obtained, the plasma state optical signal is subjected to spectral analysis to obtain a corresponding optical signal characteristic spectral line capable of reflecting the content of the nutrient element to be detected at the three-dimensional measuring point in the blade, and the content of the nutrient element to be detected at the three-dimensional measuring point of the blade is obtained by analyzing the optical signal characteristic spectral line.
On the basis of the foregoing embodiments, the obtaining of the content of the molecule to be measured in each three-dimensional measurement point on the leaf based on the fluorescence signal in the synchronous imaging detection method provided by the embodiments of the present invention specifically includes:
and obtaining a corresponding pixel point array database based on the fluorescence signal generated after each three-dimensional measuring point is excited, and obtaining the content of the molecules to be measured in each three-dimensional measuring point based on the pixel point array database.
After obtaining a fluorescence signal generated after the excitation of the molecule to be detected, performing imaging analysis on the fluorescence signal to obtain a pixel point array database capable of reflecting the content of the molecule to be detected at the three-dimensional measuring point in the leaf, and analyzing according to the pixel point array database to obtain the content of the molecule to be detected at the three-dimensional measuring point of the leaf.
On the basis of the foregoing embodiments, the optical signal characteristic spectral line in the synchronous imaging detection method provided in the embodiments of the present invention specifically includes: spectral peak position and spectral peak intensity. The optical signal characteristic spectral line for reflecting the content of the molecules to be measured at the three-dimensional measuring point in the blade comprises two characteristics of a spectral line peak position and a spectral line peak intensity, wherein the spectral line peak position represents the element type, and the spectral line peak intensity represents the content, namely the optical signal characteristic spectral line comprises the content information of the elements to be measured.
On the basis of the foregoing embodiments, the synchronous imaging detection method provided in an embodiment of the present invention further includes, before acquiring a plasma state optical signal generated by each three-dimensional measurement point on the blade after being excited by the first laser beam and a fluorescence signal generated after being excited by the second laser beam:
and establishing a three-dimensional measurement coordinate system, and determining the distribution of three-dimensional measurement points based on the cell level of the leaf. Namely, the synchronous imaging detection method provided by the embodiment of the invention further comprises the steps of establishing a three-dimensional measurement coordinate system for convenient and accurate measurement, and determining three-dimensional measurement points according to the cell levels of the leaf, wherein the leaf is provided with a plurality of cell layers, each cell contains molecules to be measured and nutrient elements to be measured, a Z axis of three-dimensional measurement is established according to the distribution of cells in different levels on the thickness, and an X-Y axis vertical to the Z axis is established on the surface of the leaf, so that the three-dimensional measurement coordinate system of the leaf is established, and the three-dimensional measurement points are uniformly selected based on the coordinate system and the cell levels of the leaf, and are all in the cell layers of the leaf.
In the embodiment of the present invention, based on the synchronous imaging detection method in the foregoing method embodiments, a synchronous imaging detection system is further provided, fig. 2 is a schematic structural diagram of the synchronous imaging detection system provided in the embodiment of the present invention, and as shown in fig. 2, the system includes a first imaging detection system 21, a second imaging detection system 23, and a main control device 22, where:
the first imaging detection system 21 acquires a plasma state optical signal generated by each three-dimensional measurement point on the blade after the first laser beam is excited, and obtains a corresponding optical signal characteristic spectral line based on the plasma state optical signal;
the second imaging detection system 23 acquires a fluorescence signal generated by each three-dimensional measurement point on the blade after being excited by the second laser beam, and obtains a corresponding pixel point array database based on the fluorescence signal;
the main control device 22 obtains the content of the nutrient element to be measured in each three-dimensional measuring point of the blade based on the optical signal characteristic spectral line corresponding to each three-dimensional measuring point, obtains the element distribution diagram of the nutrient element to be measured in the blade based on the content of the nutrient element to be measured in all the three-dimensional measuring points, obtains the content of the molecule to be measured in each three-dimensional measuring point based on the pixel lattice column database corresponding to each three-dimensional measuring point, and obtains the molecular spectrogram of the molecule to be measured in the blade based on the content of the molecule to be measured in all the three-dimensional measuring points.
Specifically, the synchronous imaging detection system provided by the embodiment of the present invention includes a first imaging detection system 21, a second imaging detection system 23, and a main control device 22, where the main control device 22 controls the first imaging detection system 21 and the second imaging detection system 23 to sequentially detect a same three-dimensional measurement point of a blade, analyzes and obtains contents of a nutrient element to be detected and a molecule to be detected in the three-dimensional measurement point in the blade, and then sequentially detects all three-dimensional measurement points of the blade. The first imaging detection system 21 is configured to obtain a plasma state optical signal generated by each three-dimensional measurement point on the blade after being excited by the first laser beam, and perform spectral analysis on the plasma state optical signal to obtain an optical signal characteristic spectral line capable of reflecting the content of the nutrient element to be detected at the three-dimensional measurement point in the blade; the second imaging detection system 23 is configured to obtain a fluorescence signal generated by each three-dimensional measurement point on the blade after being excited by the second laser beam, and perform imaging analysis on the fluorescence signal to obtain a pixel array database capable of reflecting the content of the molecule to be detected at the three-dimensional measurement point in the blade; the main control device 22 obtains the content of the nutrient element to be measured in each three-dimensional measuring point of the blade based on the optical signal characteristic spectral line corresponding to each three-dimensional measuring point, obtains an element distribution diagram capable of reflecting the spatial distribution of the molecule to be measured on the blade based on the content of the nutrient element to be measured in all the three-dimensional measuring points, obtains the content of the molecule to be measured in each three-dimensional measuring point based on the pixel lattice column database corresponding to each three-dimensional measuring point, and obtains a molecular spectrogram capable of reflecting the spatial distribution of the molecule to be measured on the blade based on the content of the molecule to be measured in all the three-dimensional measuring points, so that the content comparison distribution condition of the molecule to be measured and the nutrient element to be measured at the corresponding position in the distribution space in the blade is obtained through the.
The sequence of the first imaging detection system 21 and the second imaging detection system 23 for detecting the same three-dimensional measurement point of the blade can be selected according to specific needs, that is, the first imaging detection system 21 and the second imaging detection system 23 can simultaneously detect; after the first imaging detection system 21 detects the image, the second imaging detection system 23 may detect the image; after the second imaging detection system 23 performs the detection, the first imaging detection system 21 performs the detection, for example, taking the acquisition of the spatial distribution of nitrogen elements and chlorophyll molecules as an example, the order of the first imaging detection system 21 and the second imaging detection system 23 detecting the same three-dimensional measurement point of the leaf may be: the second imaging detection system 23 detects a first three-dimensional measurement point of the blade, after fluorescent signals generated after chlorophyll molecules in the three-dimensional measurement points are excited are obtained, the first imaging detection system 21 detects the same three-dimensional measurement point of the blade, namely the first three-dimensional measurement point, plasma state optical signals generated after nutrient elements to be detected corresponding to the three-dimensional measurement point are excited are obtained, then, after the second imaging detection system 23 detects a second three-dimensional measurement point of the blade and obtains corresponding fluorescent signals, the first imaging detection system 21 detects and detects the second three-dimensional measurement point, and the detection of all the three-dimensional measurement points is completed by the second imaging detection system 23 and the first imaging detection system 21 in sequence.
The synchronous imaging detection system provided by the embodiment of the invention sequentially detects all three-dimensional measurement points of the blade through the first imaging detection system and the second imaging detection system, obtains a plasma state optical signal generated after the nutrient element to be detected is excited and a fluorescence signal generated by a molecule to be detected, performs spectral analysis on the plasma state optical signal to obtain an optical signal characteristic spectral line capable of reflecting the content of the nutrient element to be detected, performs imaging analysis on the fluorescence signal to obtain a pixel point array database capable of reflecting the content of the molecule to be detected, and obtains the content contrast distribution conditions of the molecule to be detected and the corresponding position of the nutrient element to be detected in the distribution space in the blade through the content of the nutrient element to be detected and the content of the molecule to be detected in all the three-dimensional measurement points.
On the basis of the foregoing embodiments, the first imaging detection system in the synchronous imaging detection system provided in the embodiments of the present invention includes: first laser, first optical signal incident optical fiber, first optical system, first optical signal outgoing optical fiber and spectral analysis device, wherein:
the first laser is used for emitting a first laser beam for exciting the nutrient element to be detected to generate a plasma state optical signal and emitting the first laser beam into the first optical signal incidence optical fiber;
the first optical signal incidence optical fiber is used for guiding the received first laser beam into the first optical system;
the first optical system is used for focusing the first laser beam to a three-dimensional measuring point in the blade, exciting a nutrient element to be measured to generate a corresponding plasma state optical signal, and conducting the plasma state optical signal to enter a first optical signal exit optical fiber;
the first optical signal emitting optical fiber is used for transmitting the received plasma state optical signal to the spectral analysis device;
the spectrum analysis device is used for obtaining corresponding optical signal characteristic spectral lines based on the plasma state optical signals. That is, the first imaging detection system in the synchronous imaging detection system provided by the embodiment of the present invention includes a first laser, a first optical signal incident optical fiber, a first optical system, a first optical signal emitting optical fiber, and a spectrum analysis device, where the first laser emits a first laser beam with a specific frequency to the first optical signal incident optical fiber, the first laser beam is transmitted to the first optical system through the first optical signal incident optical fiber, and then is focused to a three-dimensional measurement point through the focusing function of the first optical system, so as to excite a nutrient element to be measured, thereby generating a plasma state optical signal, and under the focusing function of the first optical system, the plasma state optical signal is collected and transmitted to the first optical signal emitting optical fiber, and is transmitted to the spectrum analysis device through the first optical signal emitting optical fiber, the spectrum analysis device performs spectrum analysis on the obtained plasma state optical signal, and obtaining an optical signal characteristic spectral line capable of reflecting the content of the nutrient element to be measured at the three-dimensional measuring point in the leaf, wherein the spectral analysis device can select a spectrometer.
On the basis of the foregoing embodiments, the second imaging detection system in the synchronous imaging detection system provided in the embodiments of the present invention includes: second laser, second light signal incident optical fiber, second optical system, second light signal outgoing optical fiber and formation of image detection device, wherein:
the second laser is used for emitting a second laser beam for exciting the molecules to be detected to generate fluorescence and emitting the second laser beam into a second optical signal incidence optical fiber;
the second optical signal incidence optical fiber is used for guiding the received second laser beam into a second optical system;
the second optical system is used for focusing the second laser beam to a three-dimensional measuring point in the blade, exciting the molecules to be measured to generate corresponding fluorescence and transmitting the fluorescence to enter a second optical signal exit optical fiber, wherein the second optical system comprises a focusing lens with a preset angle;
the second optical signal emitting optical fiber is used for transmitting the received fluorescence to the imaging detection device;
the imaging detection device is used for obtaining a corresponding pixel point array database based on the fluorescence signal. That is, the second imaging detection system in the synchronous imaging detection system provided in the embodiment of the present invention includes a second laser, a second optical signal incident optical fiber, a second optical system, a second optical signal emitting optical fiber, and an imaging detection device, where the second laser emits a second laser beam with a specific frequency to the second optical signal incident optical fiber, the second laser beam is transmitted to the second optical system through the second optical signal incident optical fiber, and then focused to a three-dimensional measurement point through the focusing function of the second optical system, to excite a molecule to be detected, so as to generate a fluorescence signal, and the fluorescence signal is collected and transmitted to the second optical signal emitting optical fiber under the focusing function of the second optical system, and transmitted to the imaging detection device through the second optical signal emitting optical fiber, and the imaging detection device performs imaging processing on the obtained fluorescence signal, so as to obtain a pixel point array database capable of reflecting the content of the molecule to be detected at the three-dimensional measurement point in the blade, the imaging detection device can select an imaging detector, and a focusing lens with a preset angle in the second optical system can ensure that a second laser beam emitted by a second laser in the second optical system and a laser beam emitted by the second optical system are focused on the same two three-dimensional measurement points.
On the basis of the foregoing embodiments, the synchronous imaging detection system provided in the embodiments of the present invention further includes:
a digital pulse delay controller; the trigger ports of the first laser, the second laser, the imaging detection device and the spectral analysis device are all connected with the digital pulse delay controller, the output ports of the digital pulse delay controller, the imaging detection device and the spectral analysis device are all connected with the main control device, wherein:
the main control device sets the working frequency of the first laser, the working frequency of the second laser, the starting time sequence between the first laser and the spectral analysis device, the starting time sequence between the second laser and the imaging detection device and the working time sequence between the first laser and the second laser through the digital pulse delay controller;
the main control device acquires the optical signal characteristic spectral line obtained by the spectral analysis device through the output port of the spectral analysis device, and acquires an element distribution diagram of the nutrient elements to be measured in the blades based on the optical signal characteristic spectral line and the three-dimensional coordinates of the three-dimensional measurement points;
the main control device obtains the pixel point array database obtained by the imaging detection device through the output port of the imaging detection device, and obtains the molecular spectrogram of the molecules to be detected in the blade based on the pixel point array database and the three-dimensional coordinates of the three-dimensional measurement points. The main control device sets the working frequency of a first laser and the working frequency of a second laser through a digital pulse delay controller, so that a first laser beam emitted by the first laser is a laser with a specific frequency and used for exciting molecules to be detected, and a second laser beam emitted by the second laser is a laser with a specific frequency and used for exciting nutrient elements to be detected; the method comprises the steps of setting a starting time sequence between a first laser and an imaging detection device, a starting time sequence between a second laser and a spectrum analysis device and a working time sequence between the first laser and the second laser, so that the whole system can be started orderly and safely, and setting the working time sequence between the first laser and the second laser, so that the first laser and the second laser can sequentially emit laser beams to focus on the same three-dimensional measuring point according to the set time sequence and interval time. Meanwhile, the main control device also sets the exposure time of the spectral analysis device and the imaging detection device.
On the basis of the foregoing embodiments, the synchronous imaging detection system provided in the embodiments of the present invention further includes:
airtight measuring chamber, sample platform, three-dimensional moving platform and three-dimensional motor drive controlling means, as shown in the figure, wherein:
inert gas is filled into the closed measuring chamber, and the closed measuring chamber is used for placing the sample table and the three-dimensional moving platform;
the sample table and the three-dimensional moving platform are positioned in the closed measuring chamber, the sample table is placed on the three-dimensional moving platform and moves along with the three-dimensional moving platform according to the set three-dimensional coordinates, and the sample table is used for placing a sample to be measured;
the three-dimensional motor driving control device is connected with the three-dimensional moving platform and used for controlling the three-dimensional moving platform to move according to a preset three-dimensional coordinate. The volume of the sample stage can be 12mm by 8mm by 3mm, and the sample stage is provided with a device for fixing plant leaves, is fixed on the three-dimensional moving platform, is positioned at the lower part of the closed measuring chamber and is used for placing a sample to be measured; the three-dimensional moving platform can be a flat plate made of alloy aluminum and having the thickness of 15mm by 5mm, is positioned in the closed measuring chamber, and moves in one or more directions of an X axis, a Y axis and a Z axis in three directions to change the position of the sample platform;
the three-dimensional motor driving control device controls the stepping motors in the X axis direction, the Y axis direction and the Z axis direction; the volume of the sealed measuring chamber can be 30cm by 20cm, the sealed measuring chamber is made of aluminum alloy, the lower part of the sealed measuring chamber can be opened and closed, and the whole measuring chamber is subjected to confidentiality processing. When the device works, the lower part of the closed measuring chamber is opened, the three-dimensional moving platform drives the sample platform to move downwards until the sample platform is exposed outside the closed measuring chamber, then a sample to be measured is placed on the sample platform, then the three-dimensional moving platform drives the sample platform to enter the closed measuring chamber, and the lower part of the closed sample chamber is closed. Before measurement, inert gas is filled into the sealed measurement chamber through the gas charging and discharging device. The closed measuring chamber is completely isolated from the external environment and is used for isolating light rays to form a darkroom so as to avoid the interference of visible light; the inside of the closed measuring chamber is filled with inert gas, so that the influence of nitrogen in the air on nitrogen elements in the plant leaves is avoided.
To more clearly illustrate the laser focusing principle of the synchronous imaging detection system in the above embodiment, referring to fig. 3 for details, fig. 3 is a schematic laser focusing diagram of the synchronous imaging detection system according to the embodiment of the present invention, as shown in fig. 3, a first laser beam emitted by a first laser enters a first optical system through a first optical signal entrance fiber 311, the first laser beam in a horizontal state output by the first optical signal entrance fiber passes through a 45 ° mirror 36, emits a first laser beam 341 in a vertical downward direction, and is focused on a plant leaf 38 of a sample stage 37 through a focusing lens, a plasma state optical signal 342 generated by laser induction is collected into a first optical signal exit fiber 312 and is conducted to a spectrometer, wherein a spot diameter may be 0.1mm, the first laser may be a YAG laser, a wavelength may be 1064nm, a maximum energy output by a pulsed laser may be 200mj, the highest repetition frequency of the pulse can be 20Hz, the pulse width can be 3-5ns, and the emergence angle can be within 1 mrad; the spectrum range of the spectrometer can be selected from 200-1100nm, the resolution can be selected from 0.2nm, and the signal-to-noise ratio is 250: the spectrometer is used for obtaining characteristic spectral line peak position and spectral line peak position intensity information of a plasma state optical signal;
the second laser beam 321 emitted by the second laser enters the second optical system through the second optical signal incident fiber 331, the second laser beam output by the second optical signal incident fiber 331 is focused by the focusing lens 35, so that the focal point of the second laser beam coincides with the focal point of the second optical system, the second optical system focuses the second laser beam on the plant leaf 38, the laser induces to generate fluorescence, and the fluorescence signal 332 excited by the second laser beam is collected by the focusing lens, transmitted to the second optical signal exiting fiber 322 and acquired by the imaging detector. Wherein the second laser can be selected as tunable laser with spectral range of 225 nm and 2400nm, continuously adjustable, pulse width of 10ns, and laser irradiation power density of 4.25MW/cm2For generating a pulsed laser beam of variable wavelength.
Specifically, the work flow of the whole synchronous imaging detection system is further described in detail, fig. 4 is a schematic diagram of the work flow of the synchronous imaging detection system provided in the embodiment of the present invention, as shown in fig. 4, wherein the nutrient element to be detected is a nitrogen element, and the molecule to be detected is a chlorophyll molecule, and the flow includes:
step 41, the main control device sends an instruction to the three-dimensional motor driving control device, the sample platform for placing the plant leaves is moved to a measurement initial position through the three-dimensional moving platform, the laser focus is ensured to be focused on the surface of the plant leaves, and initial coordinates (0, 0, 0) are recorded;
step 42, the control device sends an instruction to set the exposure time of the imaging detector and the spectrometer, wherein the exposure time can be selected to be 0.5ms, the main control device sends an instruction to the digital pulse delay controller to set the working frequency of the first laser and the second laser, the working frequency can be set to be 0.5Hz, the delay time interval of starting between the first laser and the imaging detector, the delay time interval of starting between the second laser and the spectrometer and the time starting interval of exciting laser by the first laser and the second laser are set to be 0.6 ms;
43, emitting laser by the first laser according to the set starting time to strike plant leaves to form chlorophyll fluorescence molecules, obtaining a chlorophyll fluorescence molecule pixel point array database by the imaging detector according to the set delay time interval, and transmitting the chlorophyll fluorescence molecule pixel point array database to the main control device, wherein the imaging detector can be 10us according to the set delay time interval;
step 44, when the starting time of the second laser is reached, the second laser emits laser to strike the plant leaves to form plasma state optical signals, the spectrometer obtains characteristic spectral lines of the plant leaf plasma state optical signals according to a set delay time interval and transmits the characteristic spectral lines to the main control device, and the main control device qualitatively and quantitatively analyzes the content of nitrogen elements according to the relation between the peak position of the spectral lines and the intensity of the spectral lines, wherein the delay time interval can be 10 us;
step 45, the main control device sends an instruction to the three-dimensional motor driving control device, the sample platform for placing the plant leaves is moved to the next preset position point through the three-dimensional moving platform, the step 42 to the step 44 are repeated, the measurement of the preset point is completed, and the process is repeated until the measurement of all three-dimensional measurement points in the preset measurement area is completed;
step 46, the main control device selects 100um × 100um × 50um according to the preset measurement area, selects the set step length as S ═ 10um, and selects X, Y, Z scanning points in the axial direction as (10, 10, 5); the master control device sends the three-dimensional motor driveAnd the control device drives the sample stage to perform N-shaped scanning on an X-Y plane through the three-dimensional moving platform, and performs vertical movement in the Z-axis direction on each measuring point of the X-Y plane to complete the preset three-dimensional plane measurement. Wherein, the main control device records the three-dimensional coordinates (x) of the three-dimensional mobile platform in real timei，yi，zi) (ii) a The master control device receives the pixel point array database d output by the imaging detectoriThe main control device acquires spectral intensity information I of the spectrometeri. The master control device comprehensively considers the position information { (x)1，y2，z3)…(xi，yi，zi)…(x10，y10，z10) Determining the column of database information (d) for pixel points1…di…dn) And spectral intensity information (I)1…Ii…In) And completing the imaging analysis and the element characteristic spectrum analysis of the fluorescence pixel points of the preset measurement area so as to obtain a three-dimensional molecular spectrogram of the plant leaf and a three-dimensional nitrogen element distribution map of the plant leaf, and thus obtaining the content contrast distribution condition of the chlorophyll molecules and the nitrogen elements at corresponding positions in the distribution space in the leaf through the three-dimensional molecular spectrogram and the three-dimensional nitrogen element distribution map, wherein the three-dimensional molecular spectrogram is a three-dimensional laser-induced fluorescence hyperspectral image.
The above-described embodiments of the apparatus are merely illustrative, and the units described as separate parts may or may not be physically separate, and parts displayed as units may or may not be physical units, may be located in one place, or may be distributed on a plurality of network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the solution of the present embodiment. One of ordinary skill in the art can understand and implement it without inventive effort.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (7)
1. A method of simultaneous imaging detection, comprising:
acquiring a plasma state optical signal generated after each three-dimensional measuring point on the blade is excited by a first laser beam and a fluorescence signal generated after each three-dimensional measuring point is excited by a second laser beam;
obtaining the content of the nutrient elements to be measured in each three-dimensional measuring point on the blade based on the plasma state optical signal, and obtaining an element distribution map of the nutrient elements to be measured in the blade based on the content of the nutrient elements to be measured in all the three-dimensional measuring points; obtaining the content of molecules to be detected in each three-dimensional measuring point on the blade based on the fluorescence signals, and obtaining a molecular spectrogram of the molecules to be detected in the blade based on the content of the molecules to be detected in all the three-dimensional measuring points;
the method for acquiring the plasma state optical signal generated after the excitation of the first laser beam and the fluorescence signal generated after the excitation of the second laser beam of each three-dimensional measuring point on the blade specifically comprises the following steps:
the method comprises the steps that a first laser beam and a second laser beam are converged to a three-dimensional measuring point on a blade, and a plasma state optical signal generated after the first laser beam is excited and a fluorescence signal generated after the second laser beam is excited corresponding to the three-dimensional measuring point are obtained;
sequentially focusing all three-dimensional measuring points on the blade through a first laser beam and a second laser beam;
and the plasma state optical signal and the fluorescence signal corresponding to each three-dimensional measuring point are synchronously acquired.
2. The method according to claim 1, wherein the obtaining of the content of the nutrient element to be measured in each three-dimensional measurement point on the blade based on the plasma state optical signal specifically comprises:
and obtaining corresponding optical signal characteristic spectral lines based on the plasma state optical signals generated after each three-dimensional measuring point is excited, and obtaining the content of the nutrient elements to be measured in each three-dimensional measuring point based on the optical signal characteristic spectral lines.
3. The method according to claim 1, wherein the obtaining of the content of the molecule to be measured in each three-dimensional measurement point on the leaf based on the fluorescence signal comprises:
and obtaining a corresponding pixel point array database based on the fluorescence signal generated after each three-dimensional measuring point is excited, and obtaining the content of the molecules to be measured in each three-dimensional measuring point based on the pixel point array database.
4. The method according to claim 2, wherein the optical signal characteristic line specifically comprises: spectral peak position and spectral peak intensity.
5. The method of claim 1, wherein before acquiring a plasma state light signal generated after the excitation of the first laser beam and a fluorescence signal generated after the excitation of the second laser beam at each three-dimensional measurement point on the blade, further comprising:
and establishing a three-dimensional measurement coordinate system, and determining the distribution of three-dimensional measurement points based on the cell level of the leaf.
6. A simultaneous imaging detection system, comprising:
first formation of image detecting system, second formation of image detecting system and master control device, wherein:
the first imaging detection system is used for acquiring a plasma state optical signal generated by each three-dimensional measuring point on the blade after the first laser beam is excited, and acquiring a corresponding optical signal characteristic spectral line based on the plasma state optical signal;
the second imaging detection system is used for acquiring a fluorescence signal generated by each three-dimensional measuring point on the blade after being excited by the second laser beam, and acquiring a corresponding pixel point array database based on the fluorescence signal;
the main control device is used for obtaining the content of nutrient elements to be detected in each three-dimensional measuring point of the blade based on the optical signal characteristic spectral line corresponding to each three-dimensional measuring point, obtaining an element distribution diagram of the nutrient elements to be detected in the blade based on the content of the nutrient elements to be detected in all the three-dimensional measuring points, obtaining the content of molecules to be detected in each three-dimensional measuring point based on the pixel lattice column database corresponding to each three-dimensional measuring point, and obtaining a molecule spectrogram of the molecules to be detected in the blade based on the content of the molecules to be detected in all the three-dimensional measuring points;
the first imaging detection system comprises: first laser, first optical signal incident optical fiber, first optical system, first optical signal outgoing optical fiber and spectral analysis device, wherein:
the first laser is used for emitting a first laser beam for exciting the nutrient element to be detected to generate a plasma state optical signal and emitting the first laser beam into the first optical signal incidence optical fiber;
the first optical signal incidence optical fiber is used for guiding the received first laser beam into the first optical system;
the first optical system is used for focusing the first laser beam to the three-dimensional measuring point in the blade, exciting the nutrient element to be measured to generate a corresponding plasma state optical signal, and conducting the plasma state optical signal to enter the first optical signal exit optical fiber;
the first optical signal emitting optical fiber is used for transmitting the received plasma state optical signal to the spectrum analysis device;
the spectrum analysis device is used for obtaining corresponding optical signal characteristic spectral lines based on the plasma state optical signals;
the second imaging detection system comprises: second laser, second light signal incident optical fiber, second optical system, second light signal outgoing optical fiber and formation of image detection device, wherein:
the second laser is used for emitting a second laser beam for exciting the molecules to be detected to generate fluorescence and emitting the second laser beam into the second optical signal incidence optical fiber;
the second optical signal incidence optical fiber is used for guiding the received second laser beam into the second optical system;
the second optical system is used for focusing the second laser beam to the three-dimensional measuring point in the blade, exciting the molecules to be measured to generate corresponding fluorescence and transmitting the fluorescence to enter the second optical signal exit optical fiber, wherein the second optical system comprises a focusing lens with a preset angle;
the second optical signal emergent optical fiber is used for transmitting the received fluorescence to the imaging detection device;
the imaging detection device is used for obtaining the corresponding pixel point array database based on the fluorescent signal;
and the plasma state optical signal and the fluorescence signal corresponding to each three-dimensional measuring point are synchronously acquired.
7. The simultaneous imaging detection system according to claim 6, further comprising:
a digital pulse delay controller; the first laser, the second laser, the trigger port of the imaging detection device and the trigger port of the spectral analysis device are all connected with the digital pulse delay controller, the output port of the imaging detection device and the output port of the spectral analysis device are all connected with the master control device, wherein:
the main control device sets the working frequency of the first laser, the working frequency of the second laser, the starting time sequence between the first laser and the spectrum analysis device, the starting time sequence between the second laser and the imaging detection device and the working time sequence between the first laser and the second laser through the digital pulse delay controller;
the main control device acquires the optical signal characteristic spectral line obtained by the spectral analysis device through the output port of the spectral analysis device, and acquires an element distribution diagram of the nutrient element to be measured in the blade based on the optical signal characteristic spectral line and the three-dimensional coordinates of the three-dimensional measurement points;
the main control device obtains the pixel point array database obtained by the imaging detection device through the output port of the imaging detection device, and obtains the molecular spectrogram of the molecule to be detected in the blade based on the pixel point array database and the three-dimensional coordinates of the three-dimensional measurement points.
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