FluorCamDesktop plant multispectral fluorescence imaging system

——The most widely used instrument technology for experimental research on plant phenotype and physiological ecology

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 desktop plant multispectral fluorescence imaging system is a highly integrated, innovative, user-friendly, and widely used high-end plant in vivo imaging technology equipment. The high-sensitivity CCD lens, four fixed LED light source boards, and control system are integrated into a dark adaptation operation box (a fifth light source board can also be optionally placed on the top according to needs). Plant samples are placed on a partition in the dark adaptation operation box, which can be adjusted in seven levels of height; 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 more than 20 parameters such as 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 (optional), OJIP rapid fluorescence kinetics analysis (optional), and GFP green fluorescent protein imaging (optional), 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, the standard configuration is red and white, with 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.

The left figure A shows the Fv/Fm of cucumber leaves under 100% red light source conditions, while the left figure B shows the Fv/Fm of cucumber leaves under 30% blue light source conditions; The upper right figure shows the relationship between photosynthesis intensity and light intensity (different proportions of blue light), while the lower right figure shows the relationship between stomatal conductance and light intensity (different proportions of blue light)

· Can run chlorophyll fluorescence imaging, multispectral fluorescence imaging, GFP steady-state fluorescence imaging

· Optional TetraCam color imaging module with a maximum imaging area of 20x25cm, used for morphological imaging analysis of leaves or plants and comparative analysis of chlorophyll fluorescence imaging

· Optional hyperspectral imaging unit and infrared thermal imaging unit, digitalization and visualization of plant traits, comprehensive measurement and analysis of plant morphology, photosynthetic efficiency, biochemical traits, stomatal conductance, stress and resistance, etc

· Optional large-scale mobile plant imaging analysis system with an imaging area of 35x35cm, capable of performing chlorophyll fluorescence imaging, infrared thermal imaging, and RGB imaging analysis

Latest application case:

Hendrik KupperZuzana Benedikty and others published Plant Physiology in February 2019 Analysis of OJIP Chlorophyll Fluorescence Kinetics and QA Reoxidation Kinetics by Direct Fast Imaging， This study is the first to use the high-speed imaging sensor FluorCam desktop plant chlorophyll fluorescence imaging system and FKM multispectral microfluorescence imaging system, with imaging speeds of up to 4000fps@640x512 ， QA reoxygenation chlorophyll fluorescence kinetics imaging measurement of single pulse saturation scintillation150,000Mmol/m2.s1.

Attachment: OJIP rapid fluorescence kinetic determination analysis parameters include:

a)FoInitial fluorescence or minimum fluorescence, fluorescence at 50 μ s

b)FjFluorescence at 2ms

c)FiFluorescence at 60ms

d)POr Fm: maximum fluorescence

e)Vj=(FJ Fo)/(Fm Fo): j-order fluorescence relative variable

f)Vi=(Fi Fo)/(Fm Fo): i-order fluorescence relative variable

g)MoTRo/RC Eto/RC=4 (F300 Fo)/(Fm Fo): Initial slope of fluorescence transient, also known as the initial slope of OJIP curve

h)AreaThe area between the OJIP curve and Fm, which can be referred to as the complementary area, needs to be standardized as Sm=Area/(Fm Fo) in order to compare different samples. Sm is a measure of the energy required to close all photoreactive centers

i)Fix AreaOJIP fixed area, the area below the F value at 40 microseconds to 1 second on the OJIP curve

j)SmStandardized OJIP compensation area, reflecting multiple turnover of QA restoration

k)SsVj/Mo: Standardized OJ compensation area, reflecting single turnover QA restoration

l)N=Sm/Ss=Sm Mo(1/Vj)OJIP QA restores turnover quantity (between 0 and t)_Fm）

m)Phi­­­_Po＝QY＝φpo＝TRo/ABS＝Fv/Fm， Maximum photon yield, initial capture ratio of absorbed photon flux reaction center

n)Psi_o＝ψo＝ETo/TRo＝1-Vj， Capture the ratio of electron transfer quantum flux in the quantum flux of light

o)Phi_Eo＝φ_Eo=ETo/ABS=(1-(Fo/Fm))(1-Vj)， The ratio of absorbed photon flux to electron transport photon flux, also known as the quantum yield of electron transport at t=0

p)Phi_Do＝φ_Do＝1-φpo＝Fo/Fm， Energy dissipation and quantum yield of light (t=0)

q)Phi_pav= φpav = φpo (Sm/t)_Fm）, average photon production, t_FmTime required to achieve Fm (ms)

r)ABS/RC=Mo (1/Vj) (1/QY): is the absorbed light quantum flux per unit reaction center, where the reaction center only refers tothe active (QA to QA– reducing) centers(Same below). QY=TRo/ABS=Fv/Fm

s)TRo/RC=Mo (1/Vj): Initial (or maximum) capture of light quantum flux per unit reaction center (resulting in the reduction of QA, i.e. an increase in the reaction center closure ratio B)

t)ETo / RC=Mo (1/Vj) (1-Vj): Unit reaction center initial electron transfer light quantum flux

u)DIo / RC=(ABS/RC) - (TRo/RC): Energy loss per unit reaction center

v)ABS/CSThe absorbed light quantum flux per unit sample cross-section,CS stands for the excited cross-section of the tested sample(Same below). ABS/CSo=Fo, ABS/CSm=Fm, TRo/CSx=QY (ABS/CSx) - Capture energy or photon flux per unit cross-section

w)TRo/CSo＝QY. Fo；ETo/CSo＝φ_Eo.Fo ＝QY. (1-Vj). Fo

x)RC/CSxDensity of reaction centers,RC / CS0 (active RCs per excited cross-section)

y)PI_ABS＝(RC/ABS)(φpo/φ_Do）(ψ o/Vj): A "performance" index or survival index based on the absorbed photon flux

z)PIcs＝(RC/CSx)(φpo/φ_Do）(ψ o/Vj): Performance index or survival index based on cross-section