CN112362603A - Device for synchronously acquiring reflection spectrum and absorption spectrum images of plants at high flux and working method thereof - Google Patents
Device for synchronously acquiring reflection spectrum and absorption spectrum images of plants at high flux and working method thereof Download PDFInfo
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- CN112362603A CN112362603A CN202011523687.9A CN202011523687A CN112362603A CN 112362603 A CN112362603 A CN 112362603A CN 202011523687 A CN202011523687 A CN 202011523687A CN 112362603 A CN112362603 A CN 112362603A
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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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
Abstract
The invention relates to a device for synchronously acquiring reflection spectrum and absorption spectrum images of plants at high flux and a working method thereof, wherein the device comprises a box body, an LED area array light source, a space moving platform, an image acquisition module, a control module, an objective table, a drawing frame, an objective box and universal wheels; the control module is respectively connected with the space mobile platform and the image acquisition module; the dynamic chlorophyll fluorescence detection, the steady chlorophyll fluorescence detection and the multispectral imaging technology are combined to carry out plant physiological nondestructive detection, corresponding devices are provided, the same camera is used for obtaining three images, three signals (a dynamic chlorophyll fluorescence signal, a steady chlorophyll fluorescence signal and a multispectral reflection signal) are arranged on the same pixel point of each image, and high-flux information collection of small and medium-sized plants is met. The invention can acquire three images by using the same camera, so that the same pixel point of each image has three signals, and the plant physiological information can be acquired more comprehensively.
Description
Technical Field
The invention relates to the technical field of plant detection, in particular to a device for synchronously acquiring reflection spectrum and absorption spectrum images of plants at high flux and a working method thereof.
Background
The nondestructive acquisition and discrimination of the plant physiological condition information have great significance to the fields of breeding, automatic production and the like. At present, technologies for acquiring plant physiological information by using a nondestructive detection technology comprise a visible light multispectral imaging technology, a near-infrared imaging technology, a terahertz detection technology, a chlorophyll fluorescence imaging technology and the like, and certain achievements are obtained for detecting the plant physiological condition.
The chlorophyll fluorescence imaging technology developed on the basis of the increasingly mature chlorophyll fluorescence dynamics technology can reflect and visualize the utilization condition of the plant on the light energy, and is widely applied to the field of plant physiological state detection. At present, most chlorophyll fluorescence imaging technologies cannot get rid of the constraint of 'dark adaptation' before measurement, the light reaction and the dark reaction are disjointed after the dark adaptation, and the process of actually carrying out photosynthesis and accumulating organic matters on plants is carried out under the dynamic balance of high-speed running of the light reaction and the dark reaction. At the same time, few theoretical studies have begun to focus on the correlation between steady-state chlorophyll fluorescence kinetics changes and the photosynthesis process under photoadaptive conditions.
At present, a device prepared based on an imaging spectrum technology acquires at most two kinds of image information, the acquired images are mostly dynamic chlorophyll fluorescence images and multispectral images, steady chlorophyll fluorescence images are not involved, and the mutual relation between steady chlorophyll fluorescence change and the photosynthesis process under the condition of light adaptation is not noticed.
Disclosure of Invention
In view of this, the present invention provides a device for synchronously obtaining plant reflection spectrum and absorption spectrum images at high flux and a working method thereof, wherein different illumination modes are set by a control module, so as to simulate light intensity variation under natural conditions, enhance the difference of light responses of plants to different illumination, and realize accurate detection of plant photosynthesis efficiency.
The invention is realized by adopting the following scheme: a device for synchronously acquiring reflection spectrum and absorption spectrum images of plants at high flux comprises a large box body, an LED area array light source, a space moving platform, an image acquisition module, a control module, an objective table, a drawing frame, an objective box and universal wheels; the space moving platform comprises a space moving track assembly; the space moving track assembly comprises X, Y, Z main tracks with three directions; the whole space moving structure adopts a three-axis linkage system, a X, Y axis adopts a transmission mode of a synchronous belt and a linear guide rail pair, and a Z axis adopts a transmission mode of a ball screw pair and a linear guide rail pair;
the image acquisition module is arranged on a moving assembly in the Z-axis direction of the space moving platform and is used for acquiring a plant reflection spectrum image and an absorption spectrum image;
the control module comprises an upper computer, a singlechip module and an electric cabinet; the single chip microcomputer module comprises a master single chip microcomputer, a first slave single chip microcomputer and a second slave single chip microcomputer; the master single chip microcomputer is connected with the upper computer, and the first slave single chip microcomputer and the second slave single chip microcomputer are both connected with the master single chip microcomputer and used for controlling the master single chip microcomputer through the upper computer so as to control the first slave single chip microcomputer and the second slave single chip microcomputer; the first slave single chip microcomputer is connected with the space mobile platform and used for controlling the space mobile platform to move; the second slave single chip microcomputer is connected with the image acquisition module and used for controlling the image acquisition module; the electric cabinet comprises a motor driving module and a power supply module; the power supply module is respectively connected with the LED area array light source, the space mobile platform, the image acquisition module and the control module and used for supplying power to the modules.
Furthermore, the main track in the X-axis direction consists of a pair of guide rail sliding block mechanisms on two sides of the living platform and a group of synchronous belt transmission mechanisms for respectively driving the guide rail sliding block mechanisms, and the effective stroke is 1300 mm; meanwhile, two motors are arranged on the synchronous belt transmission mechanism; in order to ensure that the running states, the moving directions and the speeds of the sliding block mechanisms on the two sides are consistent, the two motors arranged on the synchronous belt transmission mechanism are controlled by the same first slave single chip microcomputer, and then the sliding blocks are driven to move through the transmission belt.
Furthermore, the main track in the Y-axis direction consists of two parallel guide rail sliding block mechanisms and a group of synchronous belt transmission mechanisms, and the effective stroke is 800 mm; two guide rail both sides are fixed respectively on 220 mm's aluminium alloy, and hold-in range structural mounting passes through the pinion rack with the open-loop hold-in range and the long bolt is fixed on Y, Z axle connecting plates at two guide rail intermediate positions to use the bolt to fix the connecting plate on Y axle slider, be used for the realization to turn into the rotary motion of band pulley the linear motion of slider, and then drive the mechanism of Z axle and carry out linear motion on the Y axle direction.
Furthermore, the main track in the Z-axis direction adopts a screw rod sliding block mechanism, and the effective stroke is 500 mm; the screw rod sliding block mechanism comprises a ball screw rod pair and a linear guide rail pair; the nut seat of the screw rod sliding block is connected with the image acquisition module through a rectangular iron sheet with the thickness of 2mm and the length of 560 mm.
Furthermore, the image acquisition module is a box body provided with a reflector at the side wall, and the box body is connected with the nut seat of the screw rod sliding block through a rectangular iron sheet; a light source component serving as an excitation light source, a camera shooting unit, a 10 mm lens, a filter wheel arranged on the lens, a light filter arranged on the filter wheel, a stepping motor for pushing the filter wheel to rotate, a motor supporting frame for fixing the stepping motor, a control board for placing a second slave single chip microcomputer and a closed box for accommodating plants, the excitation light source and the camera shooting unit are arranged in the box body; the control panel is connected with the light source assembly and used for controlling and changing the light source under the condition that the upper computer controls the master single chip microcomputer and further controls the second slave single chip microcomputer; the control panel is connected with the stepping motor and is used for controlling the rotation of the stepping motor under the condition that the upper computer controls the master singlechip and further controls the second slave singlechip, so that the rotation of the filter wheel is driven, and light rays emitted to the light inlet end of the camera shooting unit are filtered by using the light filter on the filter wheel; the control panel is also connected with the camera shooting unit and used for controlling the camera shooting unit to carry out image acquisition under the condition that the upper computer controls the master singlechip and further controls the second slave singlechip, and transmitting the acquired images back to the upper computer for image display and classified storage;
the shooting direction of the camera shooting unit points to the plants in the closed box; a lens is arranged in front of the camera shooting unit, and a filtering wheel is arranged between the camera shooting unit and the lens; the light source assembly can change a light source under the control of the second slave single chip microcomputer, and a 620nm red light source and a 380nm purple light source are turned on when an absorption spectrum image is collected; when the reflection spectrum image is collected, multispectral light sources with central wavelengths of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm are turned on in sequence.
Further, the camera shooting unit is a monochrome CCD camera; six working positions are arranged on the filtering wheel; the working positions comprise five filtering positions and a zero position; the zero position does not filter light; each light filtering position is provided with a light filter for filtering light; the optical filters of all the filtering positions are respectively a 680nm band-pass filter, a 440nm band-pass filter, a 520nm band-pass filter, a 690nm band-pass filter and a 740nm band-pass filter; the light source component comprises a plurality of LED excitation light sources which are arranged in a ring shape; the monochromatic CCD camera is arranged in the center of the ring where the LED excitation light sources are arranged; the plant is located under the monochrome CCD camera during measurement.
Further, the LED excitation light source comprises an actinic light source with the central wavelength of 620nm, an ultraviolet light source with the central wavelength of 400nm, and a multispectral light source with the central wavelengths of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm; the actinic light source is used for exciting dynamic chlorophyll fluorescence; the ultraviolet light source is used for exciting steady chlorophyll fluorescence; the multispectral light source is used for acquiring a multispectral reflection spectrum image.
Further, the invention also provides a working method of the device for synchronously acquiring the reflection spectrum and the absorption spectrum image of the plant based on high flux, which comprises the following steps:
step S1: placing the plant in a dark environment, and carrying out dark adaptation treatment on the plant so as to reset the photosynthetic system of the plant to an initial state;
step S2: in a dark environment, measuring light illumination, and acquiring a minimum chlorophyll fluorescence image of the plant subjected to dark adaptation treatment by a camera through a filter element; then irradiating with saturated light, and obtaining the maximum chlorophyll fluorescence image of the plant after dark adaptation treatment by a camera through a filter element;
step S3: the method comprises the steps of enabling a plant to be in an actinic light irradiation environment, irradiating the plant with actinic light for t1 duration, and obtaining a plant reflection image irradiated by the actinic light for t1 duration through a camera after the plant reaches a light adaptation state; then a camera is used for obtaining the plant dynamic instantaneous fluorescence image at the moment through a light filtering element;
step S4: turning off actinic light, turning on an ultraviolet light source, enabling the plant to be in an ultraviolet light illumination environment for t2 time, controlling the filter wheel to turn to the position of a narrow-band filter with the wavelength of 440nm by the second slave single-chip microcomputer to obtain a blue spectral fluorescence image with the plant in a stable state, controlling the filter wheel to turn to the position of a narrow-band filter with the wavelength of 520nm by the second slave single-chip microcomputer to obtain a green spectral fluorescence image with the plant in a stable state, controlling the filter wheel to turn to the position of a narrow-band filter with the wavelength of 690nm by the second slave single-chip microcomputer to obtain a red spectral fluorescence image with the plant in a stable state;
step S5: turning off an ultraviolet light source, controlling a filter wheel to turn to a zero position by a second slave singlechip, namely, turning on 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm light sources in sequence to measure multispectral images of the plant, wherein the interval of the period is 1s, and collecting red, green, blue and near-infrared multispectral reflection spectrum images in sequence;
step S6: the absorption spectrum image is obtained through steps S2 to S4, and the reflection spectrum image is obtained through step S5.
Further, the dark adaptation treatment of the plant in step S1 is performed for not less than 25 minutes, and the plant is irradiated with 740nm far-infrared light having an intensity of about 10. mu. mol. m-2. S-1 during the dark adaptation treatment.
Further, the actinic light irradiation environment in step S3 simulates a natural environment with actinic light having a light intensity varying.
Compared with the prior art, the invention has the following beneficial effects:
(1) according to the invention, different illumination modes are set through the control module, the light intensity change of natural conditions is simulated, the difference of the light response of the plants to different illumination is enhanced, and the accurate detection of the photosynthesis efficiency of the plants is realized.
(2) The signals which can be synchronously acquired by the invention comprise dynamic fluorescence, steady-state fluorescence and multispectral reflection signals, and three images can be acquired by using the same camera, so that the same pixel point of each image has three signals, and the plant information can be acquired more comprehensively.
Drawings
Fig. 1 is a schematic view of the overall structure of the apparatus according to the embodiment of the present invention, in which 0 is a large box, 1 is an LED area array light source, 2 is a space moving platform, 3 is an image acquisition module, 4 is an object stage, 5 is a drawing frame, 6 is an upper computer PC, 7 is an electric cabinet, 8 is an object carrying box, and 9 is a universal wheel.
FIG. 2 is a schematic structural diagram of a spatial mobile platform according to an embodiment of the present invention, wherein 2-1 is a main track in the X-axis direction; 2-2 is a main track in the Y-axis direction; and 2-3 is a main track in the Z-axis direction.
FIG. 3 is a general schematic view of an image capturing module 3 according to an embodiment of the present invention, wherein 3-1 is a light source module; 3-2 is a lens; 3-3 is a filtering wheel; 3-4 is an optical filter; 3-5 is a stepping motor; 3-6 is a motor support frame; 3-7 is a CCD camera; 3-8 are control panels; 3-9 is a closed box for accommodating the plants, the excitation light source and the camera shooting unit; 3-10 are reflectors.
Fig. 4 is a lamp source distribution diagram of an image acquisition module of the apparatus according to the embodiment of the present invention.
Fig. 5 is a flowchart of the operation of an embodiment of the present invention.
Detailed Description
The invention is further explained below with reference to the drawings and the embodiments.
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
As shown in fig. 1, the embodiment provides a device for synchronously acquiring reflection spectrum images and absorption spectrum images of plants at high flux, which includes a large box, an LED area array light source, a spatial mobile platform, an image acquisition module, a control module, an object stage, a drawing frame, an object box and a universal wheel; the space moving platform comprises a space moving track assembly; the space moving track assembly comprises X, Y, Z main tracks with three directions; the whole space moving structure adopts a three-axis linkage system, a X, Y axis adopts a transmission mode of a synchronous belt and a linear guide rail pair, and a Z axis adopts a transmission mode of a ball screw pair and a linear guide rail pair;
the image acquisition module is arranged on a moving assembly in the Z-axis direction of the space moving platform and is used for acquiring a plant reflection spectrum image and an absorption spectrum image;
the control module comprises an upper computer, a singlechip module and an electric cabinet; the single chip microcomputer module comprises a master single chip microcomputer, a first slave single chip microcomputer and a second slave single chip microcomputer; the master single chip microcomputer is connected with the upper computer, and the first slave single chip microcomputer and the second slave single chip microcomputer are both connected with the master single chip microcomputer and used for controlling the master single chip microcomputer through the upper computer so as to control the first slave single chip microcomputer and the second slave single chip microcomputer; the first slave single chip microcomputer is connected with the space mobile platform and used for controlling the space mobile platform to move; the second slave single chip microcomputer is connected with the image acquisition module and used for controlling the image acquisition module; the electric cabinet comprises a motor driving module and a power supply module; the power supply module is respectively connected with the LED area array light source, the space mobile platform, the image acquisition module and the control module and used for supplying power to the modules. The object box is placed on the object stage; the electric cabinet is arranged on the side wall of the large box body; the universal wheels are arranged at the lower part of the large box body; the LED area array light source is arranged on the inner side wall of the top plate on the large box body.
The upper computer is used for controlling the linkage of the space mobile platform, controlling image acquisition and real-time display and classified storage of images; the master single chip microcomputer is responsible for forwarding information, and the slave single chip microcomputer respectively controls the light source of the acquisition device and the space moving platform to move.
In this embodiment, the master single chip microcomputer, the first slave single chip microcomputer and the second slave single chip microcomputer are all single chip microcomputers with the model number of Arduino Mega 2560.
As shown in fig. 2, in this embodiment, the main track in the X-axis direction is composed of a pair of rail slider mechanisms respectively disposed at two sides of the living platform and a set of synchronous belt transmission mechanisms respectively driving the rail slider mechanisms, and the effective stroke is 1300 mm; meanwhile, two motors are arranged on the synchronous belt transmission mechanism; in order to ensure that the running states, the moving directions and the speeds of the sliding block mechanisms on the two sides are consistent, the two motors arranged on the synchronous belt transmission mechanism are controlled by the same first slave single chip microcomputer, and then the sliding blocks are driven to move through the transmission belt.
In this embodiment, the two motors mounted on the synchronous belt drive are each of a type (57BYG 250C).
As shown in fig. 2, in this embodiment, the main track in the Y-axis direction is composed of two parallel rail-slider mechanisms and a set of synchronous belt transmission mechanisms, and the effective stroke is 800 mm; two guide rail both sides are fixed respectively on 220 mm's aluminium alloy, and hold-in range structural mounting passes through the pinion rack with the open-loop hold-in range and the long bolt is fixed on Y, Z axle connecting plates at two guide rail intermediate positions to use the bolt to fix the connecting plate on Y axle slider, be used for the realization to turn into the rotary motion of band pulley the linear motion of slider, and then drive the mechanism of Z axle and carry out linear motion on the Y axle direction.
As shown in fig. 2, in the present embodiment, the main track in the Z-axis direction adopts a screw-slider mechanism, and the effective stroke is 500 mm; the screw rod sliding block mechanism comprises a ball screw rod pair and a linear guide rail pair; the nut seat of the screw rod sliding block is connected with the image acquisition module through a rectangular iron sheet with the thickness of 2mm and the length of 560 mm.
As shown in fig. 3, in this embodiment, the image acquisition module is a box body with a reflective mirror at a side wall, the box body can accommodate a plant to be measured, and the box body is connected with the nut seat of the lead screw slider through a rectangular iron sheet; a light source component serving as an excitation light source, a camera shooting unit, a 10 mm lens, a filter wheel arranged on the lens, a light filter arranged on the filter wheel, a stepping motor for pushing the filter wheel to rotate, a motor supporting frame for fixing the stepping motor, a control board for placing a second slave single chip microcomputer and a closed box for accommodating plants, the excitation light source and the camera shooting unit are arranged in the box body; the control panel is connected with the light source assembly and used for controlling and changing the light source under the condition that the upper computer controls the master single chip microcomputer and further controls the second slave single chip microcomputer; the control panel is connected with the stepping motor and is used for controlling the rotation of the stepping motor under the condition that the upper computer controls the master singlechip and further controls the second slave singlechip, so that the rotation of the filter wheel is driven, and light rays emitted to the light inlet end of the camera shooting unit are filtered by using the light filter on the filter wheel; the control panel is also connected with the camera shooting unit and used for controlling the camera shooting unit to carry out image acquisition under the condition that the upper computer controls the master singlechip and further controls the second slave singlechip, and transmitting the acquired images back to the upper computer for image display and classified storage;
the shooting direction of the camera shooting unit points to the plants in the closed box; a lens is arranged in front of the camera shooting unit, and a filtering wheel is arranged between the camera shooting unit and the lens; the light source assembly can change a light source under the control of the second slave single chip microcomputer, and a 620nm red light source and a 380nm purple light source are turned on when an absorption spectrum image is collected; when the reflection spectrum image is collected, multispectral light sources with central wavelengths of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm are turned on in sequence.
In the present embodiment, the stepper motor is of the type 24BYJ 48.
In this embodiment, the image pickup unit is a monochrome CCD camera; six working positions are arranged on the filtering wheel; the working positions comprise five filtering positions and a zero position; the zero position does not filter light; each light filtering position is provided with a light filter for filtering light; the optical filters of all the filtering positions are respectively a 680nm band-pass filter, a 440nm band-pass filter, a 520nm band-pass filter, a 690nm band-pass filter and a 740nm band-pass filter; the light source component comprises a plurality of LED excitation light sources which are arranged in a ring shape; the monochromatic CCD camera is arranged in the center of the ring where the LED excitation light sources are arranged; the plant is located under the monochrome CCD camera during measurement.
In this embodiment, the LED excitation light source includes an actinic light source with a central wavelength of 620nm, an ultraviolet light source with a central wavelength of 400nm, and a multispectral light source with a central wavelength of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm, 940 nm; the actinic light source is used for exciting dynamic chlorophyll fluorescence; the ultraviolet light source is used for exciting steady chlorophyll fluorescence; the multispectral light source is used for acquiring a multispectral reflection spectrum image. As shown in fig. 4, the LED illumination sources are distributed as follows: the inner circle is a multispectral light source and is provided with 12-wavelength LED lamps, the light sources of the outer two circles are both used for exciting plant fluorescence light sources and are 16 light sources, and the LED lamps with two wavelengths (400 nm and 620 nm) are alternately distributed to improve the uniformity of the light sources.
As shown in fig. 5, preferably, the embodiment further provides a working method of the apparatus for synchronously acquiring the reflection spectrum and the absorption spectrum image of the plant based on high throughput, which includes the following steps:
step S1: placing the plant in a dark environment, and carrying out dark adaptation treatment on the plant so as to reset the photosynthetic system of the plant to an initial state;
step S2: in a dark environment, measuring light illumination, and acquiring a minimum chlorophyll fluorescence image of the plant subjected to dark adaptation treatment by a camera through a filter element; then irradiating with saturated light, and obtaining the maximum chlorophyll fluorescence image of the plant after dark adaptation treatment by a camera through a filter element;
step S3: the method comprises the steps of enabling a plant to be in a actinic light irradiation environment, irradiating the plant with actinic light for a time duration of t1(3-5min), and obtaining a plant reflection image irradiated with actinic light for a time duration of t1(3-5min) through a camera after the plant reaches a light adaptation state; then a camera is used for obtaining the plant dynamic instantaneous fluorescence image at the moment through a light filtering element;
step S4: (after the plant is in the ultraviolet light illumination environment for t2 time, the blue spectrum fluorescence image, the green spectrum fluorescence image, the red spectrum fluorescence image and the far infrared spectrum fluorescence image of the plant are obtained by the camera through the filter element;)
Turning off actinic light, turning on an ultraviolet light source, and after the plant is in an ultraviolet light illumination environment for t2(3-5min), turning a second slave single-chip microcomputer controlled filter wheel to a 440nm narrow-band filter position to obtain a plant stable blue spectral fluorescence image, turning the second slave single-chip microcomputer controlled filter wheel to a 520nm narrow-band filter position to obtain a plant stable green spectral fluorescence image, turning the second slave single-chip microcomputer controlled filter wheel to a 690nm narrow-band filter position to obtain a plant stable red spectral fluorescence image, and turning the filter wheel to a 740nm narrow-band filter position to obtain a far infrared spectral fluorescence image;
step S5: turning off an ultraviolet light source, controlling a filter wheel to turn to a zero position by a second slave singlechip, namely, turning on 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm light sources in sequence to measure multispectral images of the plant, wherein the interval of the period is 1s, and collecting multispectral reflection spectrum images of red, green, blue, near infrared and the like in sequence;
step S6: the absorption spectrum image is obtained through steps S2 to S4, and the reflection spectrum image is obtained through step S5.
In this example, the dark adaptation treatment of the plant described in step S1 is performed for not less than 25 minutes, and the plant is irradiated with 740nm far-infrared light having an intensity of about 10. mu. mol. m-2. S-1 during the dark adaptation treatment.
In the present embodiment, the actinic light irradiation environment in step S3 simulates a natural environment with actinic light having a light intensity that varies.
Preferably, the dynamic chlorophyll fluorescence detection, the steady chlorophyll fluorescence detection and the multispectral imaging technology are combined to perform plant physiological nondestructive detection, a corresponding device is provided, the same camera is used for acquiring three images, three signals (a dynamic chlorophyll fluorescence signal, a steady chlorophyll fluorescence signal and a multispectral reflection signal) are provided for the same pixel point of each image, and high-flux information acquisition of small and medium-sized plants is met.
Preferably, in the present embodiment, as shown in fig. 1, the apparatus of the present embodiment includes an LED area array light source 1; a space moving platform 2; an image acquisition module 3; an object stage 4; a drawing frame 5; an upper computer (PC); an electric cabinet 7; a carrier box 8; and a universal wheel 9.
The whole frame of the whole equipment is built by sectional materials, the frame is coated by iron sheet plates with the thickness of 1.5mm to form a sealed lightproof environment, and the inner wall of the frame is completely sprayed with black paint and frosted, so that the influence of external environmental factors is reduced as much as possible. The top end of the device is covered and provided with the LED area array light source 1 with adjustable brightness, the switch and the brightness of the light source are controlled through the single chip microcomputer, the light source can be automatically turned on at 8 points in the morning according to the approximate change situation of the sunlight illumination intensity in the natural environment, the brightness of the light source is gradually increased from 8 points to 12 points and gradually decreased from 12 points to 18 points, and finally the light source is automatically turned off at 18 points, so that the growth environment without illumination at night is simulated.
As shown in figure 3, the image acquisition device comprises an LED illumination light source serving as an excitation light source, a lens 3-2, a filter wheel 3-3 arranged on the lens 3-2, a filter 3-4 arranged on the filter wheel 3-3, a stepping motor 3-5 for pushing the filter wheel 3-3 to rotate, a motor support frame 3-6 for fixing the motor, a CCD camera 3-7 serving as a camera unit, a control panel 3-8 for outputting a control signal to control the duty ratio of the excitation light source, a closed box 3-9 for accommodating plants, the excitation light source and the camera unit, and a reflector 3-10.
In the process of plant phenotype measurement, researchers can move the device to a light-shading position through the universal wheels 9, and then phenotype measurement is carried out on plants placed on the object stage 4. When the plant phenotype data of not co-altitude is gathered, can be according to the height of plant, select to place the objective table 4 that will hold the plant on not co-altitude's pull frame 5. The image acquisition module 3 arranged on the space moving platform 2 moves on the space moving platform 2 to scan the plants on the object stage 4 in sequence. The image acquisition device comprises an imaging system, a CCD monochrome camera 1, a lens 3-2 with 10 mm and a filter wheel 3-3. Six working positions on the filter wheel 3-3 comprise a 680nm band-pass filter 3-4, narrow bands of 440nm, 520nm, 690nm and 740nm filters 3-4 and a zero position (without the filters 3-4). 3-4 parts of a 680nm band-pass filter, 3-4 parts of a filter for dynamic fluorescence measurement, 3-4 parts of filters for narrow bands of 440nm, 520nm, 690nm and 740nm for multispectral fluorescence measurement, and a zero position (without the filter 3-4) for multispectral reflection images. Dynamic fluorescence data and stable fluorescence data are collected through a chlorophyll fluorescence collection channel, multispectral reflectivity data such as red, green, blue and near infrared are collected through a multispectral collection channel, and data analysis and classified storage are carried out through computer software.
The method for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant at high flux is shown as a process in figure 4,
(1) and (3) placing the object carrying box 8 containing the plant sample to be detected on the object carrying table 4, and carrying out dark adaptation on the plant in a dark room of the box body 0 for 25 minutes under the weak far-red light irradiation.
(2) And clicking a 'sampling start' button on the upper computer, and linking three axes (X-Y-Z axes) on the space moving platform 2 to reset the image acquisition module 3 to the initial position.
(3) Z axle screw slider mechanism on space moving platform 2 drives image acquisition module 3, slowly descends, covers the plant completely in image acquisition module 3 box 0.
(4) The control module controls the filter wheel 3-3 to turn to the position of the band-pass filter 3-4 with the wavelength of 680nm, the measuring light (the intensity is about 1 mu mol.m < -2 > s < -1 >) is turned on, the camera acquires a minimum fluorescence image, the saturated light (2000 mu mol.m < -2 > s < -1 >) is turned on after 3 seconds, and the camera acquires a maximum fluorescence image.
(5) Opening actinic light, continuously irradiating the plants by the actinic light, acquiring a dynamic instantaneous fluorescence image at the time t by the camera at the time t, opening saturated light, and acquiring a maximum fluorescence image again by the camera under the illumination condition;
(6) and (3) closing actinic light, turning on an ultraviolet light source, controlling the filter wheel 3-3 to turn to the position 3-4 of the 440nm narrow-band filter by a program to obtain a blue spectral fluorescence image, then controlling the filter wheel 3-3 to turn to the position 3-4 of the 520nm narrow-band filter by the program to obtain a green spectral fluorescence image, then controlling the filter wheel 3-3 to turn to the position 3-4 of the 690nm narrow-band filter by the program to obtain a red spectral fluorescence image, and controlling the filter wheel 3-3 to turn to the position 3-4 of the 740nm narrow-band filter by a cut-off program to obtain a far-red spectral fluorescence image.
(7) Turning off an ultraviolet light source, controlling a filter wheel 3-3 to turn to a zero position (without an optical filter 3-4) by a program, sequentially turning on light sources of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm at an interval of 1s, and sequentially collecting multispectral reflection spectrum images of red, green, blue and near infrared.
(8) And finishing primary collection, driving the image collection module 3 by the Z-axis lead screw sliding block mechanism on the space moving platform 2 to slowly ascend, and scanning and collecting images of the plants on the objective table 4 in sequence.
(9) And processing the obtained plant reflection spectrum image and absorption spectrum image, analyzing the photosynthesis efficiency of plants with different genotypes, and screening varieties with excellent photosynthesis efficiency.
In this example, the half-band widths of the four narrow-band filters 3-4, namely the narrow-band filters 3-4 with the wavelengths of 440nm, 520nm, 690nm and 740nm, are all 15 nm, and the control program and the analysis program built in the control module are written in C + +.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (10)
1. A device for synchronously acquiring reflection spectrum and absorption spectrum images of plants at high flux is characterized in that: the LED space moving platform comprises a large box body, an LED area array light source, a space moving platform, an image acquisition module, a control module, an objective table, a drawing frame, an objective box and universal wheels; the space moving platform comprises a space moving track assembly; the space moving track assembly comprises X, Y, Z main tracks with three directions; the whole space moving structure adopts a three-axis linkage system, a X, Y axis adopts a transmission mode of a synchronous belt and a linear guide rail pair, and a Z axis adopts a transmission mode of a ball screw pair and a linear guide rail pair;
the image acquisition module is arranged on a moving assembly in the Z-axis direction of the space moving platform and is used for acquiring a plant reflection spectrum image and an absorption spectrum image;
the control module comprises an upper computer, a singlechip module and an electric cabinet; the single chip microcomputer module comprises a master single chip microcomputer, a first slave single chip microcomputer and a second slave single chip microcomputer; the master single chip microcomputer is connected with the upper computer, and the first slave single chip microcomputer and the second slave single chip microcomputer are both connected with the master single chip microcomputer and used for controlling the master single chip microcomputer through the upper computer so as to control the first slave single chip microcomputer and the second slave single chip microcomputer; the first slave single chip microcomputer is connected with the space mobile platform and used for controlling the space mobile platform to move; the second slave single chip microcomputer is connected with the image acquisition module and used for controlling the image acquisition module; the electric cabinet comprises a motor driving module and a power supply module; the power supply module is respectively connected with the LED area array light source, the space mobile platform, the image acquisition module and the control module and used for supplying power to the modules.
2. The device for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant with high flux according to claim 1, is characterized in that: the main track in the X-axis direction consists of a pair of guide rail sliding block mechanisms on two sides of the living platform and a group of synchronous belt transmission mechanisms for respectively driving the guide rail sliding block mechanisms, and the effective stroke is 1300 mm; meanwhile, two motors are arranged on the synchronous belt transmission mechanism; in order to ensure that the running states, the moving directions and the speeds of the sliding block mechanisms on the two sides are consistent, the two motors arranged on the synchronous belt transmission mechanism are controlled by the same first slave single chip microcomputer, and then the sliding blocks are driven to move through the transmission belt.
3. The device for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant with high flux according to claim 1, is characterized in that: the main track in the Y-axis direction consists of two parallel guide rail sliding block mechanisms and a group of synchronous belt transmission mechanisms, and the effective stroke is 800 mm; two guide rail both sides are fixed respectively on 220 mm's aluminium alloy, and hold-in range structural mounting passes through the pinion rack with the open-loop hold-in range and the long bolt is fixed on Y, Z axle connecting plates at two guide rail intermediate positions to use the bolt to fix the connecting plate on Y axle slider, be used for the realization to turn into the rotary motion of band pulley the linear motion of slider, and then drive the mechanism of Z axle and carry out linear motion on the Y axle direction.
4. The device for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant with high flux according to claim 1, is characterized in that: the main track in the Z-axis direction adopts a screw rod sliding block mechanism, and the effective stroke is 500 mm; the screw rod sliding block mechanism comprises a ball screw rod pair and a linear guide rail pair; the nut seat of the screw rod sliding block is connected with the image acquisition module through a rectangular iron sheet with the thickness of 2mm and the length of 560 mm.
5. The device for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant with high flux according to claim 1, is characterized in that: the image acquisition module is a box body provided with a reflector at the side wall, and the box body is connected with the nut seat of the screw rod sliding block through a rectangular iron sheet; a light source component serving as an excitation light source, a camera shooting unit, a 10 mm lens, a filter wheel arranged on the lens, a light filter arranged on the filter wheel, a stepping motor for pushing the filter wheel to rotate, a motor supporting frame for fixing the stepping motor, a control board for placing a second slave single chip microcomputer and a closed box for accommodating plants, the excitation light source and the camera shooting unit are arranged in the box body; the control panel is connected with the light source assembly and used for controlling and changing the light source under the condition that the upper computer controls the master single chip microcomputer and further controls the second slave single chip microcomputer; the control panel is connected with the stepping motor and is used for controlling the rotation of the stepping motor under the condition that the upper computer controls the master singlechip and further controls the second slave singlechip, so that the rotation of the filter wheel is driven, and light rays emitted to the light inlet end of the camera shooting unit are filtered by using the light filter on the filter wheel; the control panel is also connected with the camera shooting unit and used for controlling the camera shooting unit to carry out image acquisition under the condition that the upper computer controls the master singlechip and further controls the second slave singlechip, and transmitting the acquired images back to the upper computer for image display and classified storage;
the shooting direction of the camera shooting unit points to the plants in the closed box; a lens is arranged in front of the camera shooting unit, and a filtering wheel is arranged between the camera shooting unit and the lens; the light source assembly can change a light source under the control of the second slave single chip microcomputer, and a 620nm red light source and a 380nm purple light source are turned on when an absorption spectrum image is collected; when the reflection spectrum image is collected, multispectral light sources with central wavelengths of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm are turned on in sequence.
6. The device for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant with high flux according to claim 5, is characterized in that: the camera shooting unit is a monochrome CCD camera; six working positions are arranged on the filtering wheel; the working positions comprise five filtering positions and a zero position; the zero position does not filter light; each light filtering position is provided with a light filter for filtering light; the optical filters of all the filtering positions are respectively a 680nm band-pass filter, a 440nm band-pass filter, a 520nm band-pass filter, a 690nm band-pass filter and a 740nm band-pass filter; the light source component comprises a plurality of LED excitation light sources which are arranged in a ring shape; the monochromatic CCD camera is arranged in the center of the ring where the LED excitation light sources are arranged; the plant is located under the monochrome CCD camera during measurement.
7. The device for synchronously acquiring the reflection spectrum image and the absorption spectrum image of the plant with high flux according to claim 6, is characterized in that: the LED excitation light source comprises an actinic light source with the central wavelength of 620nm, an ultraviolet light source with the central wavelength of 400nm, and a multispectral light source with the central wavelengths of 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm; the actinic light source is used for exciting dynamic chlorophyll fluorescence; the ultraviolet light source is used for exciting steady chlorophyll fluorescence; the multispectral light source is used for acquiring a multispectral reflection spectrum image.
8. The device for synchronously acquiring the reflection spectrum and the absorption spectrum of the plant based on the high flux of the claim 1 and the working method thereof are characterized in that: the method comprises the following steps:
step S1: placing the plant in a dark environment, and carrying out dark adaptation treatment on the plant so as to reset the photosynthetic system of the plant to an initial state;
step S2: in a dark environment, measuring light illumination, and acquiring a minimum chlorophyll fluorescence image of the plant subjected to dark adaptation treatment by a camera through a filter element; then irradiating with saturated light, and obtaining the maximum chlorophyll fluorescence image of the plant after dark adaptation treatment by a camera through a filter element;
step S3: the method comprises the steps of enabling a plant to be in an actinic light irradiation environment, irradiating the plant with actinic light for t1 duration, and obtaining a plant reflection image irradiated by the actinic light for t1 duration through a camera after the plant reaches a light adaptation state; then a camera is used for obtaining the plant dynamic instantaneous fluorescence image at the moment through a light filtering element;
step S4: turning off actinic light, turning on an ultraviolet light source, enabling the plant to be in an ultraviolet light illumination environment for t2 time, controlling the filter wheel to turn to the position of a narrow-band filter with the wavelength of 440nm by the second slave single-chip microcomputer to obtain a blue spectral fluorescence image with the plant in a stable state, controlling the filter wheel to turn to the position of a narrow-band filter with the wavelength of 520nm by the second slave single-chip microcomputer to obtain a green spectral fluorescence image with the plant in a stable state, controlling the filter wheel to turn to the position of a narrow-band filter with the wavelength of 690nm by the second slave single-chip microcomputer to obtain a red spectral fluorescence image with the plant in a stable state;
step S5: turning off an ultraviolet light source, controlling a filter wheel to turn to a zero position by a second slave singlechip, namely, turning on 460 nm, 520nm, 580 nm, 660 nm, 710 nm, 730 nm, 760 nm, 780 nm, 810 nm, 850 nm, 900nm and 940 nm light sources in sequence to measure multispectral images of the plant, wherein the interval of the period is 1s, and collecting red, green, blue and near-infrared multispectral reflection spectrum images in sequence;
step S6: the absorption spectrum image is obtained through steps S2 to S4, and the reflection spectrum image is obtained through step S5.
9. The device for synchronously acquiring the reflection spectrum and the absorption spectrum of the plant with high flux according to claim 8 and the working method thereof are characterized in that: the dark adaptation treatment for the plant in step S1 is performed for not less than 25 minutes, and during the dark adaptation treatment, 740nm far infrared light with intensity of about 10 μmol · m-2 · S-1 is used to irradiate the plant.
10. The device for synchronously acquiring the reflection spectrum and the absorption spectrum of the plant with high flux according to claim 8 and the working method thereof are characterized in that: the actinic light irradiation environment in step S3 simulates a natural environment with actinic light having a light intensity that varies.
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Cited By (2)
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|---|---|---|---|---|
| CN113777053A (en) * | 2021-09-10 | 2021-12-10 | 南京诺源医疗器械有限公司 | High-flux detection method and device based on quantum dot fluorescence and multispectral camera |
| CN114636474A (en) * | 2022-03-30 | 2022-06-17 | 江苏双利合谱科技有限公司 | Optical fiber based multi-spectral system structure and detection method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113777053A (en) * | 2021-09-10 | 2021-12-10 | 南京诺源医疗器械有限公司 | High-flux detection method and device based on quantum dot fluorescence and multispectral camera |
| CN114636474A (en) * | 2022-03-30 | 2022-06-17 | 江苏双利合谱科技有限公司 | Optical fiber based multi-spectral system structure and detection method thereof |
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