A wide range of photosynthetic parameters can be derived from fluorescence measurements, giving insight into the physiological state of all photosynthetically active organisms, including higher plants, mosses and ferns as well as various types of algae, phytoplankton and biofilms.
Chlorophyll fluorescence is a very sensitive indicator of photosynthesis. Quantitative information on the quantum yield of photosynthetic energy conversion is obtained by PAM fluorometry and the saturation pulse method (Schreiber 2004).
The IMAGING-PAM family is comprised of the MAXI, MINI, and MICROSCOPY IMAGING-PAM. The different versions employ the same Multi Control Unit, IMAG-CG, and the same camera can be used for different versions. This modularity permits easy and cost-effective switching between various applications and magnifications provided by the IMAGING-PAM M-Series.
The MAXI and MINI versions can be combined with the Gas-Exchange System GFS-3000 to integrate detailed spatial information on photosynthesis with exact analysis of photosynthetic carbon fixation.
Elevating Chlorophyll Fluorescence to the 3D Level
The integration of a precise 3D scanner into the MAXI version of the IMAGING-PAM fluorometer combines saturation pulse analysis with knowledge on size, number, position and angle of leaves. As a result, the effective light intensity per unit of leaf area can be considered when light effects on photosynthesis or photoinhibition are evaluated.
Recently, an advanced method to create three-dimensional plant images has been added to the MAXI version. This new configuration allows the projection of fluorescence analysis data on a three-dimensional plant image so that photosynthesis can be analysed in the context of whole plant architecture.
The 3D configuration provides a multitude of additional morphological data for structural phenotyping of plants. Together, morphological parameters and fluorometrically characterised, functional parameters form a new realm in plant phenotyping.
Chlorophyll Fluorescence Imaging
With the advance of highly sensitive CCD cameras and extremely strong light emitting diodes (LEDs), development of IMAGING-PAM fluorometers that not only measure images of chlorophyll fluorescence but are also fully competent in providing all relevant chlorophyll fluorescence parameters using the saturation pulse method has become possible. In this way, images of photosynthetic activity and its spatio-temporal variations can be obtained.
All IMAGING-PAM fluorometers provide images for 17 different parameters. The fluorescence parameter, Ft, is continuously monitored. Fo and Fm are assessed after dark adaptation, serving as reference for fluorescence quenching analysis by the saturation pulse method.
Besides Fv/Fm, the PS II quantum yield after dark acclimation, the PS II quantum yield during illumination, Y(II), and the quantum yields of regulated and non-regulated energy dissipation, Y(NPQ), Y(NO) as well as the apparent electron transport rate (ETR and PS) can be imaged.
A routine for measuring a PAR-absorptivity image is provided for the MAXI and the MINI version of the IMAGING-PAM (Abs.-image based on images of NIR and red light remission). From this parameter the apparent rate of photosynthesis is calculated, there is no need to use the commonly known PAR-absorptivity mean value of 0.84 (ETR) for all areas any more.
The parameter PS/50 is displayed in order to depict the apparent photosynthesis with the help of the false colour code that is also used for the other photosynthesis parameters.
The 3D version of IMAGING-PAM is particularly suited for precise measurements of whole above-ground parts of sample plants. It measures leaf heights exactly and provides a clear numerical description of single leaves (size, shape, angle, balance point) and entire plants (rosette radius, surface area and number of leaves).
For different applications the MAXI, MINI and MICROSCOPY versions are available in various configurations differing in optical geometries and excitation wavelengths. While blue excitation light is normally used for fluorescence imaging of higher plants and algae, red-orange excitation is required for cyanobacteria.
Measuring heads can also be equipped with special LEDs and filter sets for imaging fluorescence from reporter molecules like GFP. The use of powerful LEDs allows actinic intensities of up to 5000 μmol m-2 s-1 (depending on the M-Series member and configuration).
For the measurement of suspensions, additional filter plates are available for enhancing the image quality even with reflective surfaces. For PSII measurements under ambient light conditions, communication with our Universal Light Meter ULM-500 is possible, so that real time light values can be written directly to the IMAGING-PAM report file.
For extended analysis options, system combination adapters are available for facilitating imaging under climate control and combined with gas exchange analysis.
Multi Control Unit IMAG-CG |
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Design: | Aluminium housing featuring large built-in Li-ion battery, sockets for cable connections with CCD Camera IMAG-K6 or IMAG-K7, connectors for the MAXI and MINI MICROSCOPY Measuring Heads, an external light source (EXT) and Battery Charger 2120-N |
Microcontroller: | RISC processor |
User interface: | Windows 10 PC with ImagingWinGigE Software. 3D Version connected via GigE, keyboard operation. USB license dongle needed (included). |
Power supply: | Internal rechargeable Li-ion battery 14.4 V/6 Ah |
Power consumption: | 9W (500 mA) drawn from internal Li-ion battery |
Recharging time: | Approx. 4 hours (IMAGING-PAM turned off) via Battery Charger 2120-N |
Operating temperature: | -5 to +45°C |
Dimensions: | 25 cm x 10.5 cm x 11 cm (L x W x H) |
Weight: | 2.1 kg (incl. battery) |
Battery Charger 2120-N |
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Input: | 90 to 264 V AC, 47 to 63 Hz |
Output: | 19 V DC, 3.7 A |
Operating temperature: | 0 to 40°C |
Dimensions: | 15 cm x 6 cm x 3 cm (L x W x H) |
Weight: | 300 g |
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3D Version |
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Measuring Head – LED-Array Illumination Unit IMAG-MAX/L |
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Design: | LED-Array mounted on printed-circuit board in aluminium housing with central opening for CCD Cameras IMAG-K6 or IMAG-K7 with miniature ventilator and cable connections to Control Unit IMAG-CG and external 300 W power supply |
Light source for fluorescence excitation and actinic illumination: | 44 royal-blue 3 W Luxeon LEDs (450 nm) equipped with individual collimating optics; standard excitation intensity 0.5 μmol quanta m-2 s-1 PAR, modulation frequency 1-8 Hz; max. actinic intensity 2300 μmol m-2 s-1 PAR; max. saturation pulse intensity 5000 μmol quanta m-2 s-1 PAR |
Light sources for assessment of absorbed PAR: | 16 red LEDs (660 nm); 16 NIR LEDs (780 nm) |
Light field properties: | Vertical incidence on sample; LED distribution optimised for uniformity; at standard working distance maximal deviation from mean intensity ± 7% |
Dimensions: | 18.5 cm x 18.5 cm x 4.5 cm (L x W x H) |
Weight: | 1.3 kg (incl. cable 1.5 m long) |
External 300 W Power Supply |
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Input: | 90 to 264 V AC, 50/60 Hz |
Output: | 43 to 57 V, 5.2 A |
Operating temperature: | 0 to 40°C |
Dimensions: | 22.6 cm x 11 cm x 5.8 cm (L x W x H) |
Weight: | 1.75 kg |
Mounting – Projection unit IMAG-3D/GS |
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Design: | Aluminium frame with LED projector, LED array IMAG-MAX/L, camera IMAG-K6/MOD3D, sliding door and darkening blinds mounted. |
Sample position: | Potted plants in a working distance of 18 cm, max diameter 10 cm x 10 cm, height/focus field of 4-5 cm. Pots are fixed on manual turntable (4 positions). Viewing angle of the camera is 0° (straight down). |
Dimensions: | 41.5 cm x 24 cm x 50 cm (L x W x H without cables) |
Weight: | 11.32 kg (Including stand, projector, camera, turntable, without cables) |
Acer portable LED projector |
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Design: | Plastic housing with LED light source (WXGA native resolution and 700 lm luminous intensity) |
Focus: | Manual |
Projection: | DLP |
Power Consumption: | 75W |
Weight: | 510 g |
Dimensions: | 19 cm x 12 cm x 19 cm (L x W x H) |
Operating Temperature: | 5°C to 40°C |
Calibration Software IMAG-3D/CAL |
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Design: | Aluminium body as carrier for 2 different calibration patterns (calibration and fusion). Three different angle positions needed during calibration process. |
Dimension: | 14 cm x 14 cm x 10 cm (width x depth x height) |
Weight: | 650 g (calibration body without calibration plates) |
ImagingWinGigE |
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Minimum PC requirements: | Windows 10, Intel Core i5-5xx or higher, NVDIA® graphics (2 GB or more dedicated memory), SSD, 8GB RAM, internal Gigabit Ethernet (GigE), HDMI. The 3D part of the software can only be used under License. Therefore a USB dongle is necessary (one dongle is included with the basic instrument). |
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MAXI Version |
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LED-Array Illumination Unit IMAG-MAX/LR |
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Design: | LED-Array mounted on printed-circuit board in aluminium housing with central opening for CCD Cameras IMAG-K6 and IMAG-K7, filter plate IMAG-MAX/FR and miniature ventilator and cable connections to Control Unit IMAG-CG and External 300 W Power Supply |
Light source for fluorescence excitation and actinic illumination: | 44 red 3 W Luxeon LEDs (650 nm) equipped with individual collimating optics; standard excitation intensity 0.5 μmol quanta m-2 s-1 PAR, modulation frequency 1-8 Hz; max. actinic intensity 1900 μmol quanta m-2 s-1 PAR; max. saturation pulse intensity 3700 μmol quanta m-2 s-1 PAR |
Light sources for assessment of absorbed PAR: | 16 red LEDs (660 nm); 16 NIR LEDs (780 nm) |
Working distance: | Standard 18.5 cm for 9 cm x 12 cm image area; at 22.5 cm distance, 11 cm x 15 cm image area (mounted independently of IMAG-MAX/GS on separate stand) |
Light field properties: | Vertical incidence on sample; LED distribution optimised for uniformity; at standard working distance maximal deviation from mean intensity ± 7% |
Dimensions: | 18.5 cm x 18.5 cm x 4.5 cm (L x W x H) |
Weight: | 1.3 kg (incl. cable 1.5 m long) |
CCD Camera and Lenses IMAG-K7 |
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Design: | Black and white C-mount camera operated in 10-bit-mode at 30 frames/sec |
CCD Chip size: | 1/2″ (640 x 480 pixels) |
Interface: | GigE-Vision® |
Dimensions: | 8.64 cm x 4.4 cm x 2.9 cm (L x W x H) (without objective lens) |
Weight: | < 200g |
IMAG-K6 |
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Design: | Black and white C-mount camera operated in 10-bit-mode at 30 frames/sec, featuring 2 x 2 pixel binning |
CCD Chip size: | 2/3″ (1392 x 1040 pixel primary resolution) |
Interface: | GigE-Vision® |
Dimensions: | 8.64 cm x 4.4 cm x 2.9 cm (L x W x H) (without objective lens) |
Weight: | < 200g |
K7-MAX/Z |
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Design: | Cosmicar-Pentax zoom objective lens (F1.0/f=8-48 mm) and close-up lens, spacer ring, appropriate detector filter set and an adapter-frame for mounting CCD-camera IMAG-K7 or IMAG-K5 at increased working distance |
Dimensions: | 57 x 95 mm |
Weight: | 430 g (without adapter-frame) |
Filter Screw Size: | φ55 mm, P=0.75 mm |
Mount: | C-Mount |
Focal Length: | 8 to 48 mm |
Max Aperture Ratio: | 1:1.0 (f=8 to 28) to 1:1.2 (f=48) |
Iris Range: | F/1.0 to F/22 |
Temperature Range: | -20°C to +50°C |
K7-MAX/S |
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Design: | Cosmicar-Pentax objective lens (F1.2/f=12 mm), spacer ring and appropriate detector filter set |
Dimensions: | 30 x 35.5 mm |
Weight: | 67 g |
Filter Screw Size: | M27 x 0.5 |
Mount: | C-Mount |
Focal Length: | 12 mm |
Iris Range: | F/1.2 to F/22 |
Temperature Range: | -20°C to +50°C |
K6-MAX |
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Design: | Cosmicar-Pentax objective lens (F1.4/f=12.5 mm), spacer ring and appropriate detector filter set |
Dimensions: | 42 x 50 mm |
Weight: | 135 g |
Filter Screw Size: | M40.5 x 0.5 |
Mount: | C-Mount |
Focal Length: | 12.5 mm |
Iris Range: | F/1.4 to C |
Temperature Range: | -20°C to +50°C |
Camera Mounting Set K6-MAX/M and K7-MAX/M |
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Design: | Mounting set IMAG-K6 or IMAG-K7 camera on IMAG-MAX/L or IMAGMAX/LR. Consisting of camera holder and metal rod 15 cm length, 15 mm diameter. |
Mounting |
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Mounting Stand with Eye Protection IMAG-MAX/GS |
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Design: | Aluminium frame featuring two clamps for mounting LED-Array Illumination Unit IMAG-MAX/L or IMAG-MAX/LR; red perspex sliding hood for eye protection; removable bottom part |
Sample position: | Detached leaves, slides or petri dishes resting on x-y stage for variable positioning; special frame for defined positioning of multiwell plates; after removal of bottom part, possibility to jack up whole Mounting Stand for study of plants in pots or trays |
Dimensions: | 23.5 cm x 24.5 cm x 22.5 cm (L x W x H) |
Weight: | 3.1 kg |
IMAG-MAX/B |
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Design: | Aluminium frame featuring two clamps for mounting LED-Array Illumination Unit IMAG-MAX/L or IMAG-MAX/LR |
Sample position: | Detached leaves, slides or petri dishes resting on x-y stage for variable positioning; fixed working distance 18.5 cm in a plane perpendicular to the optical axis. |
Dimensions: | 18.8 cm x 17.8 cm x 20.4 cm (L x W x H) |
Weight: | 455 g |
ST-101 |
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Design: | Laboratory stand with wooden baseplate for mounting LED-Array Illumination Unit IMAG-MAX/L or IMAG-MAX/LR |
Dimensions: | 40 cm x 30 cm x 70 cm (L x W x H) |
Weight: | 455 g |
Windows Software ImagingWin |
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Minimum PC requirements: | 1.7 GHz Processor, 4 GB RAM, Gigabit Ethernet (GigE), Windows Vista, 7, 8 or 10 |
Features: |
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Optional |
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Optional filter plate IMAG-MAX/F (only for IMAGMAX/ L!) |
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Design: | Black-anodised aluminium plate with 44 individual blue filters to be mounted in front of collimating optics of IMAG-MAX/L |
Filter properties: | 1 mm blue-green glass filters (BG 39, Schott) blocking red transmission and passing 90% of blue light |
Dimensions: | 186 mm x 176 mm x 2.5 mm (L x W x H) |
Weight: | 180 g |
System Combination |
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Optional filter plate IMAG-MAX/F (only for IMAGMAX/ L!) |
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Design: | Adapter Plate with legs and eye protection for positioning IMAG-MAXI Head on 3010-GWK1 |
Dimensions: | 18.5 cm x 20 cm 17 cm (L x W x H) |
Weight: | 856 g |
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MINI Version |
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Measuring Head IMAG-MIN/B |
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Design: | 12 Luxeon LEDs 460 nm with individual short pass filters and collimator optics; 16 red 650 nm and 16 NIR 780 nm LEDs for measuring PAR-absorptivity; max. actinic intensity, 3700 μmol m-2 s-1; max. Saturation Pulse intensity, 8200 μmol m-2 s-1; frame at fixed working distance (7 cm); for imaging 24 mm x 32 mm sample area; suitable for use in combination with IMAG-K6 and IMAG-K7 with camera accessories (objectives). |
Dimensions: | 11.8 cm x 9,4 cm x 8.6 cm (L x W x H) |
Weight: | 552 g (incl. cable) |
Measuring Head IMAG-MIN/R |
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Design: | 12 Luxeon LEDs 620 nm with individual short pass filters and collimator optics; 16 red 650 nm and 16 NIR 780 nm LEDs for measuring PAR-absorptivity; max. actinic intensity, 2800 μmol m-2 s-1; max. Saturation Pulse intensity, 7700 μmol m-2 s-1; frame at fixed working distance (7 cm); for imaging 24 mm x 32 mm sample area; suitable for use in combination with IMAG-K6 and IMAG-K7 with camera accessories (objectives). |
Dimensions: | 11.8 cm x 9,4 cm x 8.6 cm (L x W x H) |
Weight: | 552 g (incl. cable) |
Measuring Head IMAG-MIN/GFP |
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Design: | 12 Luxeon LEDs 480 nm with individual short pass filters and collimator optics; 16 red 650 nm and 16 NIR 780 nm LEDs for measuring PAR-absorptivity; max. actinic intensity; frame at fixed working distance (7 cm); for imaging 24 mm x 32 mm sample area; only suitable for use in combination with IMAG-K6 with camera accessories K6-MIN, K6-MIN/FS and K6-MIN/M. |
Dimensions: | 11.8 cm x 9,4 cm x 8.6 cm (L x W x H) |
Weight: | 552 g (incl. cable) |
CCD Camera and Lenses IMAG-K7 |
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Design: | Black and white C-mount camera operated in 10-bit-mode at 30 frames/sec |
CCD Chip size: | 1/2″ (640 x 480 pixels) |
Interface: | GigE-Vision® |
Dimensions: | 8.64 cm x 4.4 cm x 2.9 cm (L x W x H) (without objective lens) |
Weight: | < 200g |
IMAG-K6 |
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Design: | Black and white C-mount camera operated in 10-bit-mode at 30 frames/sec featuring 2 x 2 pixel binning |
CCD Chip size: | 2/3″ (1392 x 1040 pixel primary resolution) |
Interface: | GigE-Vision® |
Dimensions: | 8.64 cm x 4.4 cm x 2.9 cm (L x W x H) (without objective lens) |
Weight: | < 200g |
K7-MIN |
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Design: | Cosmicar-Pentax objective lens (F1.4/f=16 mm), spacer ring and appropriate detector filter set |
Dimensions: | 30 x 33 mm |
Filter Screw Size: | M27 x 0.5 |
Mount: | C-Mount |
Focal Length: | 16 mm |
Iris Range: | F/1.4 to 22 |
Temperature Range: | -20°C to +50°C |
Weight: | 58 g |
K6-MIN |
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Design: | Cosmicar-Pentax objective lens (F1.4/f=25 mm), spacer ring and appropriate detector filter set |
Dimensions: | 30 x 37.3 mm |
Filter Screw Size: | M27 x 0.5 |
Mount: | C-Mount |
Focal Length: | 25 mm |
Iris Range: | F/1.4 to 22 |
Temperature Range: | -20°C to +50°C |
Weight: | 76 g |
K6-MIN/FS |
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Design: | For parallel imaging of GFP- and Chl-fluorescence of identical sample areas (replaces detector filter set of Camera Accessory K6-MIN). Featuring alternative detector filters for green or red fluorescence (special green filter set 500-575 nm and red filter RG665) |
Dimensions: | 42 x 50 mm |
Weight: | 135 g |
Filter Screw Size: | M40.5 x 0.5 |
Mount: | C-Mount |
Focal Length: | 12.5 mm |
Iris Range: | F/1.4 to C |
Temperature Range: | -20°C to +50°C |
Camera Mounting Set K7-MIN/M and K6-MIN/M |
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Design: | Devices for mounting camera IMAG-K6 or IMAG-K7 to the Imaging MINI-Heads |
Weight: | 166 g |
Mounting |
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IMAG-S |
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Design: | Fine drive laboratory stand with high performance rack-and-pinion drive (50 mm traverse path) for adjustment of working distance; platform base with covered 9.3 cm central hole for mounting optional sample holders |
Weight: | 3.34 kg |
IMAG-MIN/ST |
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Design: | Fine drive tripod adapter for mounting a MINI-Head onto a tripod head with UNC 1/4-20 screw threads, High performance rack-and-pinion drive (120 mm traverse path) for adjustment of working distance |
IMAG-MIN/BK |
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Design: | Leaf clip mountable on sample frame of all MINI-Heads; MINI-Head holding grip |
Weight: | 184 g |
Software |
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Windows Software ImagingWin |
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Minimum PC requirements: | 1.7 GHz Processor, 4 GB RAM, Gigabit Ethernet (GigE), Windows Vista, 7, 8 or 10 |
Features: |
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Optional |
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ST-1010 | Design: Compact Tripod, for tripod heads with UNC 1/4-20 screw threads. |
System Combination |
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IMAG-MIN/GFS |
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Design: | Adapter plate with snap-on-mount for connecting IMAG-MINI Head to Standard Measuring Head 3010-S |
Dimensions: | 9.5 cm x 6 cm x 1.4 cm (L x W x H) |
Weight: | 30 g |
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MICROSCOPY Version |
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Measuring Head IMAG-L470M |
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Design: | Microscope LED Lamp Module 470 nm (blue) for fluorescence excitation of Chl fluorescence of most algae groups. Emission peak at 470 nm. Shipped together with a set of neutral grey filters for using the system together with higher magnifications. |
Measuring Head IMAG-L625M |
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Design: | Microscope LED Lamp Module 625 nm (red-orange) for fluorescence excitation of Chl fluorescence of most algae groups including cyanobacteria. Emission peak at 625 nm. Shipped together with a set of neutral grey filters for using the system together with higher magnifications. |
Measuring Head IMAG-RGB |
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Design: | Red-Green-Blue Microscopy LED Lamp allowing computer-assisted deconvolution of major algae groups. Fluorescence excitation and actinic illumination using red (620 nm), green (520 nm), blue (460 nm) or white light (mixed 620, 520 and 460 nm); featuring fluid light guide (100 cm length, 3 mm active dia.), connecting to collimator optics at excitation port of epifluorescence microscope; with cable to be connected to RGB output socket at IMAG CG; featuring printed circuit board with separate drivers for RGB LEDs; |
CCD Camera IMAG-K6 |
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Design: | Black and white C-mount camera operated in 10-bit-mode at 30 frames/sec featuring 2 x 2 pixel binning |
CCD Chip size: | 2/3″ (1392 x 1040 pixel primary resolution) |
Interface: | GigE-Vision® |
Dimensions: | 8.64 cm x 4.4 cm x 2.9 cm (L x W x H) (without objective lens) |
Weight: | < 200g |
Microscope – IMAG-AXIOSCOPE |
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Design: | Modified AxioScope.A1 Microscope (Zeiss) adapted for IMAGING PAM applications. Comprises binocular fototube (30°/23 100:0/0:100), condenser 0.9/1.25H and transmitted light unit HAL 50. Detector filter RG665, dichroic mirror 420-640 nm, video adapter 60N-C 2/3″ 0.5x and standard lens Fluar 20x are already mounted. |
Software |
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Windows Software ImagingWin |
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Minimum PC requirements: | 1.7 GHz Processor, 4 GB RAM, Gigabit Ethernet (GigE), Windows Vista, 7, 8 or 10 |
Features: |
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Optional |
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IMAG-AX-REF |
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Design: | Reflector Module with filter set for additional microscope LED lamp modules. Consisting of a beam splitter filter (420- 640 nm) and a detector filter (665 nm), mounted in a Zeiss reflector module frame. |
Schreiber, U. (2004). Pulse-Amplitude-Modulation (PAM) Fluorometry and Saturation Pulse Method: An Overview. In G. C. Papageorgiou & Govindjee (Eds.), Chlorophyll a Fluorescence: A Signature of Photosynthesis (pp. 279–319). https://doi.org/10.1007/978-1-4020-3218-9_11