FluorCam Closed Chlorophyll Fluorescence Imaging System

PSIProfessor Nedbal, Chief Scientist of the company, and Dr. Trtilek, President of the company, were the first to combine PAM chlorophyll fluorescence technology with CCD technology. In 1996, they successfully developed and produced the FluorCam chlorophyll fluorescence imaging system worldwide (Heck et al., 1999); Nedbal et al., 2000; Govindjee and Nedbal, 2000）。 FluorCam chlorophyll fluorescence imaging technology became an important breakthrough in chlorophyll fluorescence technology in the 1990s, allowing scientists to enter the two-dimensional and microscopic worlds of photosynthesis and chlorophyll fluorescence research. At present, PSI Company has become the world's most authoritative, widely used, comprehensive, and published professional manufacturer of chlorophyll fluorescence imaging

The upper left image shows the FluorCam chlorophyll fluorescence imaging technology designed by Nedbal et al. in the 1990s (Photosynthesis Research, 66:3-12, 2000), and the right image shows the lemon color image and chlorophyll fluorescence imaging image (Photosynthetica, 38:571-579, 2000)

FluorCamThe closed chlorophyll fluorescence imaging system is a highly integrated, innovative, easy-to-use, and widely used high-end chlorophyll fluorescence technology equipment. The high-sensitivity CCD lens, four fixed LED light source boards, and control system are integrated into a dark adaptation operation box. Plant samples are placed on a partition in the dark adaptation operation box, which can be adjusted in height by 7 levels; The light source is powered by a high stability power supply unit, and four high-energy, high stability LED light source boards uniformly illuminate the plant sample, with an imaging area of up to 13×13 cmThe control system is connected to the computer via USB and controls and collects analysis data through the FluorCam software program. Suitable for plant tissues such as leaves and fruits, whole plants or multiple cultivated plants, lower plants such as mosses and lichens, algae, etc. It is widely used in plant research including algae photosynthetic physiology and ecology, plant stress physiology and susceptibility, stomatal function, plant environment such as soil heavy metal pollution response and biological detection, plant resistance detection and screening, crop breeding, phenotyping, etc.

Main functional features:

· The system is integrated into the dark adaptation operation box, which is easy to operate and mobile. It can be used for dark adaptation imaging measurement and analysis both in the laboratory and outdoors

·High sensitivity CCD lens with a time resolution of 50 frames per second, capable of quickly capturing chlorophyll fluorescence transients and imaging an area of 13x13cm

· It is the only high-end chlorophyll fluorescence technology equipment in the world that can perform OJIP rapid fluorescence dynamic imaging analysis. It can obtain the OJIP rapid chlorophyll fluorescence dynamic curve and 26 parameters including Mo (initial slope of OJIP curve), OJIP fixed area, Sm (measurement of energy required to close all light reaction centers), QY, PI (Performance Index), etc

· It is the only high-end chlorophyll fluorescence technology equipment in the world that can perform QA re oxidation kinetics imaging analysis. It can operate single cycle saturated light flash (STF) chlorophyll fluorescence induction dynamics, with a light intensity of100µsIt can reach 120000 µ mol (photons)/m ². s

· The most comprehensive and editable chlorophyll fluorescence experimental program (Protocols), including snapshot mode, Fv/Fm, Kautsky induction effect, 2 chlorophyll fluorescence quenching analysis (NPQ) protocols (2 customized light supply schemes), LC light response curve, PAR absorption and NDVI imaging analysis, QA re oxidation kinetics analysis, OJIP rapid fluorescence kinetics analysis, and GFP green fluorescent protein imaging, etc

· Automatic repeated imaging measurement analysis can be performed, with a pre-set experimental protocol, measurement frequency, and interval. The system will automatically cycle the imaging measurement and automatically store the data in the computer according to the time date (with timestamp); Two experimental protocols can also be pre-set; For example, making the system automatically run Fv/Fm during the day and NPQ analysis at night

·Equipped with a dual color chemical light excitation source, standard configuration includes red and white, and optional dual band photochemical light such as red and blue. Dual color chemical light can be used in different proportions to experiment with the photosynthetic benefits of crops/plants under different light qualities

· Optional TetraCam color imaging module with a maximum imaging area of 20 × 25cm, used for morphological imaging analysis of leaves or plants and comparative analysis of chlorophyll fluorescence imaging

Technical parameters:

· The measurement light is 617nm modulated red light, consisting of two arrays of 144 LEDs, with a duration adjustable from 10 µ s to 100 µ s;

· Bichromatic light, standard configuration includes 2 red lights+2 white lights, optional 2 red lights+2 blue lights or other wavelength light source combinations, Actinic1 light intensity300µmol(photons)/m².s， Actinic2 light intensity 2000 µ mol (photons)/m ². s; The maximum photochemical intensity can be upgraded to 3000 µ mol (photons)/m ². s. Bichromatic chemical light can be used in different proportions to experiment with the photosynthetic benefits of crops/plants under different light qualities

· The saturation light intensity can reach 4000 µ mol (photons)/m ². s, which can be upgraded to 6000 µ mol (photons)/m ². s,QAThe single cycle saturated photoflash STF for re oxidation analysis can reach 120000 µ mol (photons)/m ². s

· Photochemical light and saturation light flicker consist of two pairs of high-intensity and high stability LED light source boards, each consisting of an 8 × 9 LED array. In addition, there is a top dual color light source (740nm infrared light source and 660nm red light source) for PAR absorption and NDVI imaging measurement; High intensity and high stability LED provides a continuous, stable, and uniform light source, without causing problems such as unstable light source or short lifespan due to the use of a large number of strong and weak LEDs (such as hundreds). Using a large number of low light LEDs to compensate for the shortcomings of each LED can increase the system error rate, and any LED problem can cause instability or even render the system unusable

· Measurement parameters: Fo, Fo’, Fs, Fm, Fm’, Fp, FtDn, FtLn, Fv, Fv'/ Fm'，Fv/ Fm ,Fv',Ft,ΦPSII, NPQ_Dn, NPQ_Ln, Qp_Dn, Qp_Ln, qN, qP，QY, QY_Ln, Rfd, ETR Waiting for over 50 chlorophyll fluorescence parameters, PAR absorption, and NDVI plant spectral reflectance indices, each parameter can display a 2D fluorescence color image

· Equipped with a complete automatic measurement program (protocol), the automatic measurement program can be freely edited

a) Fv/FmMeasurement parameters include Fo, Fm, Fv, QY, etc

b) KautskyInduction effect: Fluorescence parameters such as Fo, Fp, Fv, Ft_Lss, QY, Rfd, etc

c) Fluorescence quenching analysis: Fo, Fm, Fp, Fs, Fv, QY, Φ_II， More than 50 parameters including NPQ, Qp, Rfd, qL, etc

d) Light response curve LC: Fluorescence parameters such as Fo, Fm, QY, QYLn, ETR, etc

e) PARAbsorption and NDVI (optional)

f) QARedox kinetics (optional)

g) GFPStatic fluorescence imaging measurement (optional)

h) OJIPRapid Fluorescence Dynamics Analysis (optional): 26 parameters including Mo (initial slope of OJIP curve), OJIP fixed area, Sm (measurement of energy required to close all photoreaction centers), QY, PI, etc

• high sensitivityTOMI-1CCDSensor:

a) Image resolution: 720 × 560 pixels

b) Time resolution: 50 frames per second

c) A/DConversion resolution: 12 bits (4096 grayscale levels)

d) Pixel size: 8.6 µ m × 8.3 µ m

e) Communication mode: Gigabit Ethernet

§ High resolution TOMI-2 CCD sensor (optional)

a) Line by line scanning CCD

b) Maximum image resolution: 1360 × 1024 pixels

c) Time resolution: up to 20 frames per second at the highest image resolution

d) A/DConversion resolution: 16 bits (65536 grayscale levels)

e) Pixel size: 6.45 µ m × 6.45 µ m

f) Operation mode: 1) Dynamic video mode, used for measuring chlorophyll fluorescence parameters; 2) Snapshot mode, used for measuring fluorescent proteins such as GFP and fluorescent dyes

g) Communication mode: Gigabit Ethernet

· Imaging area: 13×13cmIt can perform imaging analysis on plant leaves, plant tissues, algae, mosses, lichens, whole plants or multiple plants, 96 well plates, 384 well plates, etc

· 7Position filter wheel, chlorophyll fluorescence filter, PAR absorption and NDVI imaging measurement filter, other filters can be selected as needed (optional)

· QARe oxidation kinetics imaging analysis (optional): STF fluorescence kinetics analysis measurement can be performed, with a single cycle light flash (STF) intensity of up to 120000 µ mol (photons)/m ². s in 100 µ s

· OJIPFast Fluorescence Dynamics Module (optional): With a time resolution of 1 µ s, it can measure and analyze the OJIP curve and more than twenty related parameters, including Fo, Fj, Fi, P or Fm, Vj, Vi, Mo, Area, Fix Area, Sm, Ss, N (QA reduction turnover quantity), Phi....... Po, Psiuo, Phi-Eo, Phi-Do, Phiepav, ABS/RC (absorbed light quantum flux per unit reaction center), TRo/RC (initial captured light quantum flux per unit reaction center), ETO/RC (initial electron transfer light quantum flux per unit reaction center), DIo/RC (energy loss per unit reaction center). ABS/CS (absorption photon flux per unit sample cross-section), TRo/CSo, RC/CSx (reaction center density) PI_ABS(Performance index or survival index based on absorbed photon flux), PIcs (Performance index or survival index based on cross-section)

FluorCamChlorophyll fluorescence imaging analysis software features: Live (real-time testing), Protocols (customized experimental program selection), Pre processing (imaging preprocessing), Result (imaging analysis results) and other functional menus

· Customized experimental program protocols for customers: can set time (such as measurement light duration, photochemical light duration, measurement time, etc.), light intensity (such as photochemical light intensity of different qualities, saturation light flash intensity, modulated measurement light, etc.), with dedicated experimental program language and scripts. Users can also freely create new experimental programs using the wizard program template in the Protocol menu

· Automatic measurement and analysis function: An experimental program (Protocol) can be set to automatically perform unmanned cyclic imaging measurements. The number of repetitions and interval time can be customized by the customer, and the imaging measurement data will be automatically stored in the computer according to the time and date (with a timestamp)

· Snapshot mode: Through snapshot imaging mode, the light intensity, shutter time, and sensitivity can be freely adjusted to obtain clear and prominent steady-state and instantaneous fluorescence images of plant samples

· Imaging preprocessing: The program software can automatically identify multiple plant samples or regions, or manually select regions of interest (ROI). The shape of manual selection can be square, circular, any polygon, or sector. The software can automatically measure and analyze the fluorescence kinetics curves and corresponding parameters of each sample and selected area, with no limit on the number of samples or areas (>1000)

· Data analysis mode: It has "signal calculation and re averaging" mode (arithmetic mean) and "signal average and re calculation" mode. In high signal-to-noise ratio situations, the "signal calculation and re averaging" mode is selected, and in low signal-to-noise ratio situations, the "signal average and re calculation" mode is selected to filter out errors caused by noise

· Output results: High time resolution fluorescence dynamic image, fluorescence dynamic change video, fluorescence parameter Excel file, histogram, imaging images with different parameters, fluorescence parameter list for different ROIs, etc

· Dark adaptation operation box, built-in light source, CCD lens, green wave wheel and filter, control unit, heat dissipation device, etc., convenient for dark adaptation operation. The sample platform is 36x30cm, with a height of 7 levels adjustable. The maximum height of the sample (whole plant) can reach 12cm

· Lighting system: static or dynamic (sinusoidal)

· Spectral response:540 nmThe quantum efficiency is highest at 70%, with a 50% decrease at 400 nm and 650 nm

· Read out the noise:Below 12eRMS, typical 10e

· Full trap capacity:Greater than 70000 e (unbound)

· Bios：Firmware upgradable

· Communication method:Gigabit Ethernet

· Size:471 mm(W)×473 mm (D)×512 mm (H)

· Weight:Appr. 40 kg

· Power input:Appr. 1100 W

· Supply voltage:90–240 V

Configuration composition:

1. Host system, including dark adaptation operation box, built-in LED light source, green wave wheel and filter, CCD lens, control unit, height adjustable sample partition, heat dissipation device, etc

2. High stability power converter

3. FluorCamSystem control and data analysis software

4. Notebook computer

Place of Origin:Czech PSI

Attachment:Other FluorCam chlorophyll fluorescence imaging systems

1. FluorCamPortable photosynthetic chlorophyll fluorescence imaging system: can be used in combination with LCProSD photosynthesis analyzer, Licor6400 photosynthesis analyzer, etc

2. FluorCamPortable chlorophyll fluorescence imaging system: imaging area of 3.5x3.5cm, equipped with dark adaptation leaf clamp and multifunctional lightweight tripod, can be used for laboratory or field measurement and monitoring

3. FluorCamPortable Chl/GFP fluorescence imaging system: an extended version of the portable fluorescence imaging system that can simultaneously perform chlorophyll fluorescence imaging analysis and GFP green fluorescent protein imaging analysis

4. FluorCamClosed chlorophyll fluorescence imaging system: LED light source, CCD fluorescence monitoring lens, control unit, etc. are integrated into the dark adaptation operation box to form a complete host system. It is the only chlorophyll fluorescence imaging system in the world that can perform QA re oxidation kinetics and OJIP measurement analysis, with an imaging area of 13x13cm

5. FluorCamClosed Chl/GFP fluorescence imaging system: an extended version of the closed chlorophyll fluorescence imaging system, which can simultaneously perform chlorophyll fluorescence imaging analysis and GFP green fluorescent protein imaging analysis

6. FluorCamOpen chlorophyll fluorescence imaging system: modular, highly scalable, freely selectable with different excitation light sources and corresponding filters for imaging analysis of dynamic and steady-state chlorophyll fluorescence, adjustable lens height, imaging area of 13x13cm

7. FluorCamOpen large-scale chlorophyll fluorescence imaging system: imaging area can reach 20x20cm

8. FKMMultispectral Fluorescence Dynamic Microscopic Imaging and Spectral Analysis System: Multi excitation light, multispectral fluorescence imaging and spectral analysis, can perform microscopic imaging analysis and spectral analysis of chlorophyll fluorescence dynamics, QA re oxidation, OJIP rapid fluorescence dynamics, as well as microscopic imaging analysis of GFP fluorescence, cell fluorescence staining, etc

9. FluorcamMobile large-scale chlorophyll fluorescence imaging system: The large-scale chlorophyll fluorescence imaging platform is installed on a wheeled bracket for easy movement. The imaging platform can move up and down, with an imaging area of 35x35cm

10. FluorCamSample strip scanning chlorophyll fluorescence imaging system: The large imaging platform can scan and image the sample strip on a support of 100-500cm, with a standard scanning area length of 400cm. The imaging platform can accurately locate and automatically scan along the sample strip, and can be optionally equipped with RGB true color scanning imaging to achieve chlorophyll fluorescence imaging and true color imaging analysis

11. FluorCamMultispectral Fluorescence Imaging System: It belongs to the category of multi excitation luminescence and multispectral fluorescence imaging systems. It can not only image and analyze chlorophyll fluorescence, but also image and analyze F440 (blue fluorescence), F520 (green fluorescence), F690 (red fluorescence), and F740 (infrared fluorescence) excited by UV ultraviolet light for comprehensive research and detection of plant stress and resistance. There are three models: standard configuration, extended configuration, and large configuration

12. PlantScreenChlorophyll fluorescence and RGB true color automatic scanning imaging system: It is the basic version of the PlantScreen series high-throughput plant phenotype imaging analysis system, which can image and analyze chlorophyll fluorescence and plant RGB true color to analyze and detect the functional and morphological phenotypes of plants. The automatic scanning range is 60x129cm, and it locates and timing to obtain 4-dimensional (XYZ three-dimensional position information and time information) measurement data