Chlorophyll fluorescence imaging analysis system capable of realizing double-sided imaging

Abstract

The chlorophyll fluorescence imaging analysis system comprises a light isolation box (1), a top CCD detector (2), a side CCD detector (3), a top optical filter (4), a side optical filter, a top excitation light source (5), a side excitation light source, a detachable light isolation plate (6), a control module (8) and a computer. The imaging system carrying the double-sided light source and the double CCD detectors is used in cooperation with the detachable light isolation plate, the effect of double-sided imaging can be achieved without moving plants, user operation is simplified, and meanwhile the result accuracy is improved. The system is simple in structure and convenient to operate, and can be used as a plant physiological photosynthetic phenotype analysis instrument for stress resistance research, mutant screening and phenotype analysis of laboratory plants or crops.

Description

Technical Field

[0001] The present invention relates to the technical field of plant physiological measurement devices, and further relates to a chlorophyll fluorescence imaging analysis system capable of double-sided imaging. Background Art

[0002] Chlorophyll fluorescence is an important technique for studying the photochemical efficiency and heat dissipation force in the photosynthesis process of plants. This technique, together with a photosynthesis meter based on the gas exchange principle, explains the photosynthesis occurring in the chloroplasts of plants, crops, and algae, and can be widely applied to the research fields of plant photosynthetic physiological status, stress resistance research, mutant screening, etc. Given the wide range of its application fields, the measurement requirements are also more diverse. For example, when plant leaves are lush or the leaf inclination angle of the leaves is large, taking a fluorescence image of a single perspective cannot fully illustrate the physiological state of the plant. Therefore, it is sometimes extremely necessary to take fluorescence images of plants from multiple angles. Most of the existing chlorophyll fluorescence imaging devices place the CCD detector on the top of the light-shielding box. When taking fluorescence imaging pictures of different perspectives of the same potted plant, it is necessary to rotate the angle of the plant or even tilt the plant, which not only destroys the soil state and growth state of the plant to be measured, is extremely inconvenient to operate, but also makes the measurement repeatability poor and the accuracy low due to the operation of changing the position of the plant while the CCD detector remains unchanged.

[0003] Therefore, it is necessary to design a chlorophyll fluorescence imaging analysis system with dual CCD detectors, dual light sources, capable of multi-angle shooting and double-sided imaging. Summary of the Invention

[0004] In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is that most of the existing chlorophyll fluorescence imaging devices place the CCD detector on the top of the light-shielding box, resulting in the need to rotate the angle of the plant or even tilt the plant when taking fluorescence imaging pictures of different perspectives of the same potted plant, which not only destroys the soil state and growth state of the plant to be measured, is extremely inconvenient to operate, but also makes the measurement repeatability poor and the accuracy low due to the operation of changing the position of the plant while the CCD detector remains unchanged.

[0005] To achieve the above object, the present invention provides a chlorophyll fluorescence imaging analysis system capable of dual-sided imaging, including a light-tight box 1, a top CCD detector 2, a side CCD detector 3, a top filter 4, a side filter, a top excitation light source 5, a side excitation light source, a detachable light-tight plate 6, a control module 8 and a computer. The top CCD detector 2 is installed on the top of the light-tight box 1, the side CCD detector 3 is installed on the side of the light-tight box 1, the top filter 4 is installed at the front end of the top CCD detector 2, the side filter is installed at the front end of the side CCD detector 3, the top excitation light source 5 is installed on the top of the light-tight box 1, the side excitation light source is installed on the side of the light-tight box 1 and on the same side as the side CCD detector 3, and the detachable light-tight plate 6 is detachably installed on the top or side of the light-tight box 1, and the detachable light-tight plate 6 is located in front of the filter.

[0006] Further, slots are respectively opened on the top and side of the light-tight box 1, and the detachable light-tight plate 6 can be inserted into the slots on the side or the upper part.

[0007] Further, it further includes a lifting platform 7, and the lifting platform 7 is installed at the lower part of the light-tight box 1 for placing plants and can adjust the height of the plant placement to a suitable imaging position.

[0008] Further, the excitation light source includes an excitation light source board, and the excitation light source board includes a blue light excitation light source, a red light excitation light source and a far red light excitation light source, and a hole for embedding a CCD detector is left at the central position thereof.

[0009] Further, the maximum light intensity that the blue light excitation light source can reach on a plane 50 cm away from the light source board is 4800 μmol / m 2 / s, and the adjustable light quantum flux range of the blue light excitation light source through the control module is 0 - 100%.

[0010] Further, the control module 8 can switch between two sets of CCD detectors and excitation light source systems on the top and side, and regulate the light quantum flux magnitude and illumination duration of the excitation light source.

[0011] Further, the control module 8 can adjust the height of the lifting platform.

[0012] Further, the control module 8 is embedded and installed in the upper part of the light-tight box 1.

[0013] Further, a reflective film is pasted on the inside of the light-tight box 1 and the detachable light-tight plate to ensure a uniform light field light intensity on the imaging plane. The imaging range with a uniform light field is a plane of 30 cm * 30 cm, and the uniform difference degree of the light intensity can be controlled within ±8%.

[0014] Furthermore, the computer can acquire and analyze fluorescence images and basic parameters, including the minimum fluorescence value under dark adaptation, the maximum fluorescence value under dark adaptation, the steady-state fluorescence value under light, the maximum fluorescence value under light, and the absorbance under light. The computer can calculate photochemical efficiency parameters and fluorescence quenching parameters, including PSII photochemical quantum efficiency, photochemical quenching coefficient, and non-photochemical quenching coefficient.

[0015] Technical Effects

[0016] 1. Through innovative structural design, the present invention installs a CCD detector and a light source board on both the side and the top simultaneously, achieving the effect of double-sided imaging, effectively compensating for the defect that the chlorophyll fluorescence imaging devices on the market can only collect fluorescence imaging pictures from a single perspective. When shooting with the top light source, the light-shielding plate with a reflective film is inserted into the side card slot; when shooting with the side light source, the light-shielding plate with a reflective film is inserted into the top card slot. Fluorescence imaging parameters and pictures of the same plant are collected from multiple perspectives, avoiding data deviation caused by angle problems and ensuring the accuracy of chlorophyll fluorescence parameters.

[0017] 2. Through innovative structural design of the light source, the excitation light source selects a light source board embedded with several blue, red, and far-red light beads. One light source board is placed on the top of the light-shielding box, and the other is placed on the side of the light-shielding box. There are holes for embedding the CCD detector in the middle of the light source board. The surrounding of the light-shielding box is covered with a reflective film to increase the reflectivity of specific wavelength light, enabling most of the incident light to be reflected back, reducing light loss, and improving the resolution, brightness, and uniformity of the optical system.

[0018] 3. Through innovative structural design, a lifting platform is built in the light-shielding box, and the height of the lifting platform is adjusted by the control module to place the sample to be measured at a suitable imaging height, effectively compensating for the defects of poor flexibility and low efficiency of manually adjusting the height of the sample partition board in the chlorophyll fluorescence imagers on the market. At the same time, the lifting platform device makes the CCD detector have a wider application range, including a series of samples from sliced tissues, leaves to potted plants.

[0019] The present invention uses an imaging system equipped with double-sided light sources and double CCD detectors, used together with a detachable light-shielding plate, which can achieve the effect of double-sided imaging without moving the plant, simplifying the user operation while improving the accuracy of the results. This system has a simple structure and convenient operation, and can be used as a plant physiological photosynthetic phenotype analysis instrument for stress resistance research, mutant screening, phenotype analysis, etc. of laboratory plants or crops.

[0020] The following will further illustrate the concept, specific structure, and technical effects of the present invention with reference to the drawings to fully understand the purpose, features, and effects of the present invention. Description of the Drawings

[0021] Figure 1 It is the front right view of the overall structural schematic diagram of a chlorophyll fluorescence imaging analysis system capable of double-sided imaging, which is a preferred embodiment of the present invention;

[0022] Figure 2 It is the bottom view of the overall structural schematic diagram of a chlorophyll fluorescence imaging analysis system capable of double-sided imaging, which is a preferred embodiment of the present invention. Specific embodiments

[0023] The following introduces multiple preferred embodiments of the present invention with reference to the accompanying drawings of the specification to make its technical content clearer and easier to understand. The present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text.

[0024] In the drawings, components with the same structure are denoted by the same numerical labels, and components with similar structures or functions everywhere are denoted by similar numerical labels. The size and thickness of each component shown in the drawings are arbitrarily shown, and the present invention does not limit the size and thickness of each component. To make the illustration clearer, the thickness of some parts in the drawings is appropriately exaggerated.

[0025] As Figure 1 、 2 shown, in view of the problems existing in the prior art, the present invention provides a chlorophyll fluorescence imaging analysis system capable of double-sided imaging, including a light-tight box 1, a top CCD detector 2, a side CCD detector 3, a filter 4, an excitation light source 5, a detachable light-tight plate 6, a lifting platform 7, a control module 8, and a computer. Among them, the top CCD detector 2 is installed on the top of the light-tight box 1, the side CCD detector 3 is installed on the side of the light-tight box 1, the filter 4 is installed at the front ends of the CCD detectors 2 and 3, two excitation light sources 5 are respectively installed on the top and side of the light-tight box 1, the lifting platform 7 is installed at the lower part of the light-tight box 1, and the control module 8 is embedded in the upper part of the light-tight box 1.

[0026] The light-tight box 1 is used to provide a dark environment for plants, exclude the fluorescence signals induced by background light, and only monitor the changes in the fluorescence signals induced by pulsed light modulation light, ensuring the accuracy of fluorescence imaging parameters. The CCD detectors 2 and 3 use 1388*1038 CCD CAT6 CCD detectors, and high-precision and high-resolution chlorophyll fluorescence image data can be obtained at an appropriate distance. The filter 4 uses a long-pass filter with a wavelength of 665nm ± 6nm, which can ensure that the CCD detector detects the chlorophyll fluorescence signal.

[0027] The two excitation light sources 5 provide blue LED excitation light, and the maximum light intensity that can be achieved on the plane 50 cm away from the light plate is 4800 μmol / m 2 / s, adjustable from 0 - 100% by the control module, to provide measuring light (for measuring the minimum fluorescence and maximum fluorescence values under dark adaptation), actinic light (for measuring steady-state fluorescence), and saturating flashes (for measuring the maximum fluorescence under light). The excitation light source board also provides red light and far-red light LED excitation light for detecting absorbance, and the far-red light source can also be used to accelerate the recovery of the plant's state under dark adaptation.

[0028] The detachable light-shielding plate 6 can be inserted into the lower part of the excitation light source at the top of the light-shielding box or the right part of the excitation light source and CCD detector on the left side, for uniforming the light intensity of the light field. The reflective film is pasted on the four sides inside the light-shielding box and the detachable light-shielding plate, so that all surfaces except the excitation light source plane and the imaging plane are pasted with reflective films, ensuring that the imaging plane has a uniform light field light intensity. The imaging range with a uniform light field is a plane of 30cm * 30cm. The uniformity difference of the light intensity can be controlled within ±8%.

[0029] The height of the sample can be adjusted by the lifting table 7 to ensure that the sample is in the best fluorescence imaging position.

[0030] The control module 8 can freely switch to connect the excitation light source and CCD detector at the top or the excitation light source and CCD detector on the side and automatically trigger the CCD detector to collect chlorophyll fluorescence parameters and images. It can also adjust the magnitude of the light quantum flux of the excitation light source and the length of the illumination time. In addition, the control module can adjust the height of the lifting table.

[0031] The computer can acquire and analyze fluorescence pictures and basic parameters, such as the minimum fluorescence and maximum fluorescence values under dark adaptation, the steady-state fluorescence value, maximum fluorescence value, absorbance, etc. under light; calculate photochemical efficiency parameters and fluorescence quenching parameters, such as PSII photochemical quantum efficiency, photochemical quenching coefficient, and non-photochemical quenching coefficient, etc.

[0032] Usage method:

[0033] Step 1: Power on the system to turn on the preheating of the chlorophyll fluorescence imaging device so that it can be used normally.

[0034] Step 2: Prepare the sample to be measured, open the light-shielding box 1, and place the fully dark-adapted sample on the measuring table.

[0035] Step 3: Select the perspective to be photographed. If you need to collect pictures from the perspective of the top CCD detector, insert the light-shielding plate 6 into the card slot on the side of the box body; if you use side shooting, insert the light-shielding plate 6 into the card slot on the top of the box body. Adjust the height of the lifting table 7, and use the display screen to view and adjust the position of the sample to make the sample in the best fluorescence imaging position.

[0036] Step 4: Connect and turn on the top CCD detector 2 or the side CCD detector 3 through the control module 8. Turn on the excitation light source 5 to measure the light mode. The CCD detector automatically captures and updates the real-time image, and takes a saturated flash to obtain the minimum and maximum fluorescence values under dark adaptation; turn on the actinic light mode through the control module 8 and adjust the light intensity level. After 1 minute, take a saturated flash to obtain the steady-state fluorescence and maximum fluorescence values under light, and complete the acquisition of basic fluorescence parameters.

[0037] Step 5: Use a computer to analyze the obtained basic fluorescence parameters, calculate parameters such as light efficiency parameters and fluorescence quenching coefficients, and output them in an EXCEL table. The corresponding folder also stores the original chlorophyll fluorescence image and the false-color image.

[0038] The preferred specific embodiments of the present invention have been described in detail above. It should be understood that those of ordinary skill in the art can make many modifications and variations according to the concept of the present invention without creative labor. Therefore, all technical solutions that can be obtained by those skilled in the art in the technical field of the present invention based on the concept of the present invention through logical analysis, reasoning, or limited experiments on the basis of the prior art should be within the protection scope determined by the claims.

Claims

1. A chlorophyll fluorescence imaging analysis system capable of double-sided imaging, characterized in that: The invention comprises a light-isolating box (1), a top CCD detector (2), a side CCD detector (3), a top optical filter (4), a side optical filter, a top excitation light source (5), a side excitation light source, a detachable light-isolating plate (6), a control module (8) and a computer. The top CCD detector (2) is mounted on the top of the light-isolating box (1), the side CCD detector (3) is mounted on the side of the light-isolating box (1), the top optical filter (4) is mounted on the front end of the top CCD detector (2), the side optical filter is mounted on the front end of the side CCD detector (3), the top excitation light source (5) is mounted on the top of the light-isolating box (1), the side excitation light source is mounted on the side of the light-isolating box (1) and on the same side as the side CCD detector (3), the detachable light-isolating plate (6) is detachably mounted on the top or side of the light-isolating box (1), and the detachable light-isolating plate (6) is located at the front end of the optical filter.

2. A chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: The top and side of the light-isolating box (1) are respectively provided with card slots, and the detachable light-isolating plate (6) can be inserted into the card slots on the side or top.

3. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: It also comprises a lifting platform (7), which is installed at the lower part of the light-isolating box (1) and is used for placing plants and can adjust the placement height of the plants to a suitable imaging position.

4. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: The excitation light source comprises an excitation light source board, and the excitation light source board comprises a blue light excitation light source, a red light excitation light source and a far-red light excitation light source, and a hole for embedding a CCD detector is reserved at the center of the excitation light source board.

5. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 4, characterized in that: The maximum light intensity that the blue light excitation light source can reach at a plane 50 cm away from the light source plate is 4800 μmol / m 2 / s, the blue light excitation light source can adjust the light quantum flux range to 0-100% through the control module.

6. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: The control module (8) can switch the two sets of CCD detectors and excitation light source systems on the top and the side, and regulate the light quantum flux and illumination duration of the excitation light source.

7. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 3, characterized in that: The control module (8) can adjust the height of the lifting platform.

8. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: The control module (8) is embedded in the upper part of the light-isolating box (1).

9. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: A reflective film is attached to the interior (1) of the light-isolating box and the detachable light-isolating plate to ensure that the imaging plane has a uniform light field intensity. The imaging range of the uniform light field is a plane of 30cm*30cm, and the uniformity of the light intensity can be controlled within ±8%.

10. The chlorophyll fluorescence imaging analysis system capable of double-sided imaging as claimed in claim 1, characterized in that: The computer can obtain and analyze fluorescence images and basic parameters, including the minimum fluorescence value under dark adaptation, the maximum fluorescence value under dark adaptation, the steady-state fluorescence value under light, the maximum fluorescence value under light, and the absorbance under light. The computer can calculate photochemical efficiency parameters and fluorescence quenching parameters, including PSII photon quantum efficiency, photochemical quenching coefficient, and non-photochemical quenching coefficient.

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