FluorCam Closed GFP/Chl. Fluorescence Imaging System

FluorCam Closed GFP/Chl. Fluorescence Imaging System

FluorCamThe closed GFP/Chl. fluorescence imaging system can provide services for a range of molecular and cellular biology studies applied to green fluorescent proteins. A completely software controlled and electrically driven filtering wheel, as well as a series of filter groups, are installed on the closed fluorescence imaging system, which can detect and image GFP, EGFP, wtGFP, YFP, BFP or other bands of fluorescent proteins and fluorescein.

The FluorCam closed GFP/Chl. fluorescence imaging system can also be used to detect the dynamic changes and spatial distribution of chlorophyll fluorescence emitted by plants, measure Kautsky effect processes, fluorescence quenching, and other transient fluorescence processes (transients), and provide 2D fluorescence images. The specially designed hardware configuration enables this system to automatically convert into fluorescent protein, fluorescein imaging, or chlorophyll fluorescence detection imaging. The system is highly integrated and can perform dark adaptation of samples.

Application areas:

·Fluorescence detection of fluorescent proteins and fluorescein

·Gene transfer and biosensor technology

·Research on genetically modified plants

·Screening for resistance to different genotypes

·Research on the Interaction between Plants and Microorganisms

·Research on the Interaction between Plants and Protozoa

·Screening of plant photosynthetic characteristics and metabolic disorders

·Detection of biotic and abiotic stress

·Research on Plant Stress Resistance or Susceptibility

·Study on non-uniformity of stomata

·Research on metabolic disorders

·Growth and yield assessment

Functional features:

·Easy to operate, adjustable sample height for measurement, integrated with a dedicated dark adaptation box, convenient for dark adaptation processing and operation of the tested plants

·High resolution CCD with a resolution of up to 1392 × 1040 pixels, capable of quickly and accurately capturing chlorophyll fluorescence and GFP

·Automatic repeated imaging measurements can be performed, and an experimental program (Protocols) can be set to automatically cycle imaging measurements. Imaging measurement data is automatically stored in the computer according to the time date (with a timestamp)

·Equipped with various universal experimental protocols such as Kautsky induction effect and fluorescence quenching analysis, measuring and analyzing over 50 parameters

·Optional TetraCam color imaging module with a maximum imaging area of 20 × 25cm, used for measuring and analyzing leaf or plant morphology

Typical sample:

• Leaves, stems, seeds, roots, whole plant

• Callus tissue, seedlings in a culture dish

• Fruits and flowers

• Cyanobacteria, green algae, bacteria

• transgenic animal

Technical parameters:

• 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, 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/Fm: Measurement parameters include Fo, Fm, Fv, QY, etc

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

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

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

e) PAR absorption and NDVI (optional)

f) A Re oxidation kinetics (optional)

g) GFP and other static fluorescence imaging measurements

h) OJIP rapid fluorescence kinetic 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 resolution TOMI-2 CCD sensor

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/D conversion 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

• Light source board: 4 large high-intensity packaged LED light source boards, each light source board is composed of an 8 × 9 LED array, and the effective area of the light source board is the same as the imaging area of 13 × 13cm

• Measurement light: Standard 617nm red light, other bands optional, duration adjustable from 10 µ s to 100 µ s

• Bichromatic light: 2 red lights (617nm)+2 blue lights (470nm), maximum light intensity of Actinic1 is 300 µ mol (photons)/m ². s, maximum light intensity of Actinic2 is 2000 µ mol (photons)/m ². s; The maximum photochemical intensity can be upgraded to 3000 µ mol (photons)/m ². s

• Saturated flicker: maximum light intensity of 4000 µ mol (photons)/m ². s, upgradable to 6000 µ mol (photons)/m ². s

• Imaging area: 13 × 13cm, capable of imaging analysis of plant leaves, plant tissues, algae, mosses, lichens, whole plants or multiple plants, 96 well plates, 384 well plates, etc

• GFP measurement module: 7-bit filtering wheel and dedicated filter for measuring GFP (green fluorescent protein)

• PAR absorption measurement module (optional): Far red light 740nm (FAR), 660nm dual color LED light source board and dedicated filter, used for measuring PAR absorption and NDVI

• QA in 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

• OJIP Rapid Fluorescence Dynamics Module (optional): With a time resolution of 1 µ s, it can measure and analyze OJIP curves 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_, Psi, Phi_, Phi_, Phi_, Phi_, Phi_, Phi_, 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 (absorbed light quantum flux per unit sample cross-section), TRo/CSo, RC/CSx (reaction center density), PIABS (performance index or survival index based on absorbed light quantum flux), PIcs (performance index or survival index based on cross-section)

• UV LED light source board (optional): 365nm or 385nm, 1) measure multi-color fluorescence (see FluorCam multispectral fluorescence imaging system for details); 2) Measurement of wtGFP (wild-type green fluorescent protein), DAPI (4 ', 6-diamidino-2-phenylindole, a fluorescent dye)

• Blue LED light source board (optional): 470nm, 1) Measure EGFP (Enhanced Green Fluorescent Protein); 2) Used for the study of stomatal function

• Green LED light source board (optional): 530nm, measuring YFP (yellow fluorescent protein)

*Corresponding filters need to be selected for measuring fluorescent proteins or dyes such as wtGFP, DAPI, EGFP, YFP, etc

• FluorCam chlorophyll fluorescence imaging analysis software features: Live (live testing), Protocols (customized experimental program selection), Pre processing (imaging preprocessing), Result (imaging analysis results) and other menu functions

• 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

• CCD detector bandwidth: 400-1000nm

·Light giving system: static or dynamic (sinus style)

·Spectral response: The quantum efficiency is highest at 540nm (70%), and decreases by 50% at 400nm and 650nm

·Read out noise: below 12eRMS, typical 10e

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

·Bios: Firmware upgradable

·Communication method: Gigabit Ethernet

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

·Weight: Approx. 40 kg

·Power input: Appro. 1100 W

·Supply voltage: 90-240 V

Typical applications:

The GFP imaging image obtained by the Shanghai Institute of Life Sciences using FluorCam measurement shows that the plants emitting bright colors in the image are those expressing GFP. The more red the color is, the more GFP it expresses

NDVI and GFP imaging analysis of button coral polyps during different contraction expansion phases (T1=0, T2=30, T3=60) (Miguel, 2014)

Origin: Europe