The Z125 GFP Closed FluorCam can serve for a number of applications using green fluorescent proteins in molecular and cellular biology.  The basic system of the Z100 Closed FluorCam (CCD camera, four fixed LED panels) is supplemented by a fully motorized and software-controlled filter wheel and appropriate filter sets for the detection and imaging of green fluorescent protein (GFP), red-shifted GFP, or yellow fluorescent protein (YFP).  The blue emitting and red fluorescent protein can be detected and imaged in absence of chlorophyll.  The device configuration also provides user-friendly and quick transition between the GFP and chlorophyll fluorescence detection and imaging.

Applications:

• GFP detection and expression studies.
• Screening of photosynthetic performance.
• stress resistance studies.
• Plant-microbe interactions research.
• Bacterial-protozoan interactions research.

Samples:

• Leaves, stems, seeds, roots whole small plants, fruits
• small animals
• calluses, seedlings on petri plates
• Cyanobacteria, green algae, bacteria.
• Sample size up to 13 cm x 13 cm.

GFP Arabidopsis seedlings on petri plate:

Experiments and Measured Parameters:

• Fv/Fm
• Quenching.
• Kautsky induction.
• QA reoxidation (needs optional accessories).
• OJIP (needs optional accessories).
• Fast fluorescence induction with 1µs resolution (needs optional accessories).

Key Features:

• Combines imaging of GFP and chlorophyll fluorescence.
• Included high-resolution CCD camera: full resolution 1392 x 1040 (15 fps), 30 fps at resolution 640 x 480; integration times from microseconds to seconds; binning options. The camera is specifically intended for detection of weak signals demanding long integration times; the setup retains most of the functionalities for chlorophyll fluorescence kinetics measurement.
• Four light-upgraded LED panels (4 panels 470 nm OR 2 panels 447 nm + 2 panels 470 nm OR 2 panels 627 nm and 2 panels 470 nm) Additional UV, 505, 570, 605, 630, 735 nm (optional).
• 8 position filter wheel – motorized and software-controlled (included).
• Adjustable shelf system for different plant sizes (included).
• Dark room for adaptation (included).
• Supplied with a high-end computer and user-friendly software (included).

Filter sets for the most commonly used GFP proteins:

Proteins of the GFP family demonstrate a great spectral diversity and they often have different excitation and emission spectra. Therefore it is important to use the most appropriate filter sets so as to obtain maximum sensitivity for GFP viewing.

• Red-shifted variants ( EGFP)

Single excitation peak is centered at about 488 nm and the emission peak is at 507 nm.
Suggested filter set: short pass 480 for excitation and 532/28 or 530/25 for emission.

• Original  GFP variants (GFP)

Single excitation peak is centered at about 395 nm and the emission peak is at 504 nm.
Suggested filter set: short pass 420nm for excitation and 532/28 or 530/25 for emission.

• Blue-emitting variants (EBFP)

Excitation peak around 384 nm and the emission peak near 448 nm.
Suggested filter set:  short pass 400nm for excitation and 469/35 for emission.

Tsien (1998). Annu. Rev. Biochem. 67: 509-544.

FluorCam 7.0 Software

• Automated experimental protocols via a Windows Wizard.
• Multiple (automatically repeated) experiments.
• Automated labeling of individual plants, or samples, within the field of view.
• Kinetic analysis of data from all samples within the field of view.
• Numerous image manipulation tools.
• Imaging masks for 384-well plate, 96-well plate, Petri dish, etc.
• Barcode reader support.
• Export to Excel.
• Windows 2000, XP, Windows 7, Vista compatible.

GFP expression in corn kernels as captured by GFP FluorCam – closed:

Corn kernels captured by the GFP FluorCam – closed.  GFP expressing kernels in upper rows, control kernels in bottom rows:

The Z450 Handy GFP Cam is designed for the investigation of green fluorescent protein (GFP) expression in small plants, leaves, animals, calluses and seedlings on plates, tissues or bacterial colonies both in the laboratory and the field.  Typical size of an investigated object is 3.5 x 3.5 cm or less.  Samples are placed within the field of view of the CCD camera, and fluorescence emission is monitored quantitatively.  Values are displayed numerically and are also used to construct a false colour image of the sample to show heterogeneity of the emission signal.   With modifications this fluorcam can also be used for chlorophyll fluorescence detection and imaging like the Z300 Handy FluorCam.  For imaging of  larger areas please see the  Z125 GFP FluorCam-Closed.

• Measuring unit consists of ultrahigh sensitivity CCD camera, objective, and a sample chamber with four integrated panels of LEDs. The measuring unit is portable and perfectly suited for field operation.
• Battery pack: GFP–FluorCam needs external NiMH batteries to operate. Small switching power supply for laboratory operation and battery recharging is provided. The batteries and the power supply fit inside a convenient shoulder bag.
• CCD camera is equipped with a F1/1.4 objective with zoom. It produces 512 × 512 pixel images of 12-bit  grey scale with a maximal frequency of 50 frames / sec. Images are recorded synchronously with measuring light flashes. Data transmission is facilitated via USB2.0 port.  Optional, high resolution CCD camera is available, 1392×1040 pixels (15 frames per minute) or 640×480 pixels (30 frames per minute).
• Light Sources: Measuring flashes and the continuous actinic light are generated in four panels, each containing 25 blue LEDs (max = 455 nm) or two blue LED panels and two cyan panels (475 nm). The LEDs generate measuring flashes of 10 μs -250 μs duration as well as continuous light of adjustable intensity (max. 2000 μmol photons m^-2 s^-1) and duration. The irradiance of the measuring flashes as well as of the actinic light at the sample level is uniform within ± 5 % over an area of 3.5 × 3.5 cm.
• Software: The instrument is controlled by a Windows 2000/XP, Vista compatible FluorCam software.  The easy-to-use software features various sophisticated mathematical operations for enhanced image contrast, noise filtering and many others: automated experimental protocols via a windows wizard, multiple experiments, intelligent image segmentation, automated labeling of samples (optional), kinetic analysis of data, image manipulation tools, export to Excel

• Computing: High-end notebook computer with USB 2.0 interface and the software are included.

Image of Arabidopsis seedlings on petri plate captured by GFP-cam.  GFP-expressing seedlings in red and WT seedlings in gray:

The Z450 Handy GFP-Cam is typically produced with 455 nm or 475 nm excitation LEDs and 510 nm emission filters. This combination is best suited for the popular EGFP (red-shifted variant).

Corn Kernels captured by the Z450 Handy GFP-Cam.  GFP containing kernels are in upper rows (lighter) and control kernels (no GFP expression) in bottom rows.