Walz HEXAGON-IMAGING-PAM Chlorophyll Fluorescence and PAM Fluorometry

General Features of the Walz HEXAGON-IMAGING-PAM
In the development of the HEXAGON-IMAGING-PAM, Walz followed the desires of customers and have, in addition to an enlarged surface, also revised the L.E.D. control to create space in the technical specifications for new applications.
The HEXAGON-IMAGING-PAM housing was also adapted to the versatile applications of our customers and it allows for the measurement of leaves, algae in vessels and benthic organisms but also whole potted plants or plant trays without fussy or long adaptation times.
Safety has also been taken into consideration and the strong light pulses with which PAM instruments work should not harm the eyes of the experimenter. Walz has integrated a safety shutdown into the HEXAGON-IMAGING-PAM which suppresses the strong light intensity should the door be opened during an experiment.
Once the software is started, the HEXAGON-IMAGING-PAM works in a 2x2 pixel binning mode on an area of 20 x 24 cm.
A special zoom function allows the user to zoom to a freely selectable quarter of the measuring area without changing image resolution. This allows to work with a higher magnification on a smaller measuring area without loss of sensitivity.

Chlorophyll fluorescence is a very sensitive indicator for photosynthesis. The Quantitative information on the quantum yield of photosynthetic energy conversion is obtained by PAM fluorometry techniques and the (Schreiber U (2004) saturation pulse method  Pulse-Amplitude-Modulation (PAM) Fluorometry and Saturation Pulse Method.
A wide range of photosynthetic parameters can be obtained from fluorescence measurements. This gives insight into the physiological state of all photosynthetically active organisms, including higher plants, mosses and ferns as well as various types of phytoplankton  biofilms and algae.

Chlorophyll Fluorescence Imaging:
With the advance of highly sensitive CCD cameras, followed by modern CMOS type camera chips together with extremely strong light emitting diodes (LED), the development of IMAGING-PAM fluorometers has become possible to not only measure images of chlorophyll fluorescence but are alsototally competent in providing the relevant chlorophyll fluorescence parameters, using saturation pulse methods.
Therefore images of photosynthetic activity and their spatio-temporal variations can be obtained.
In recent years the trend has increasingly been towards very low-noise CMOS cameras offering high sensitivities. L.E.D. technology is also making continuous progress so that high irradiation intensities may be achieved using lower power.

The heating of LEDs however, leads to a change in their performance. With the new HEXAGON-IMAGING-PAM, we are focused on compensating for these power changes and therefore opening up new dimensions of measurement accuracy in imaging and illumination homogeneity. This also opens a door for new measurement methods to be integrated into the application software.

All Walz IMAGING-PAM fluorometers provide images for 17 different parameters. The fluorescence parameter Ft is continuously monitored and kinetic values can also be exported with a clock speed of 150 ms . "Fo" and "Fm" are assessed after dark adaptation, serving as reference for fluorescence quenching analysis by the  method of pulse saturation.

Besides Fv/Fm, we can image:
- PS II quantum yield after dark acclimation
- PS II quantum yield during illumination, Y(II)
- quantum yields of regulated and non-regulated energy dissipation
- Y(NPQ), Y(NO) and the apparent electron transport rate (ETR)

EXE-File PC Software ImagingWinGigE V2.57q39 (Windows 10 and above, 64-bit systems only) (45.8 MB)

Features and Graphical User Interface

The HEXAGON-IMAGING-PAM System is controlled fully by the dedicated ImagingWinGigE software.
The ImagingWinGigE software opens with the image window that occupies most of the user surface showing the Ft value as starting parameter.
Up to 100 areas of interest (AOIs) can be placed and the user may switch between parameters that shall be shown.
The values are represented in a false color scale ranging from black (0.0) to white (1.0) with red, orange yellow, blue and violet to purple in between. At first a central standard AOI is already present after the start of the software.
Different shapes can be defined and the ImagingWinGigE software also offers a sample recognition function. AOIs` positions can be moved by the new Edit function.
Additional tabs are available for further settings and pre-programmed runs or the report tables. While working in these tabs, the image window is disengaged and is always present next to the normal ImagingWin window for a better overview.
We have tried to structure the user interface in a practical way and have adopted many of the proven controls from the ImagingWin of the M-series units.

The customer may choose between 18 different parameters (Ft, Fo, Fm, F, Fm’, Fv/Fm, Y(II), Y(NPQ), Y(NO), PS/50, Abs, Red, NIR, NPQ/4, qN, qP, qL, Inh.) that can be displayed in the image window in different color modes.
In this tab the alteration of the parameters can be observed in real-time during the experiment.
The kinetics window shows various parameter values for some or all AOIs of the currently chosen experiment plotted versus time. It serves for the evaluation of dynamic dark / light phenomena (Kautsky curve or Induction curve).
Some of the possible experiments are already preset in the Kinetics or the Light Curve tab so that also the beginner finds an easy starting point for his first successful experiments.
For advanced users it is also possible to program script files with more complex structure and even remote control the device by using the software interface.
Easy light calibration using the ULM-500 Light Meter & Logger.
ImagingWinGigE in communication with the ULM-500 provides an automated light calibration routine to generate a calibrated internal light list and furthermore offers to follow an external illumination (an appropriate PAR sensor like the Walz LS-C is necessary).
Some new features can be provided solely for the ImagingWinGigE software for HEXAGON-IMAGING-PAM, not for the ImagingWin software suitable for GigE devices or the older FireWire camera versions of the IMAGING-PAM M-Series.

Control Mini-Computer IMAG-HEX/PC

The particularly powerful Intel NUC Mini-PC works very quietly and energy-efficiently.
Due to its small design, it can be perfectly combined with the HEXAGON IMAGING PAM.
It provides a fast Ethernet interface, HDMI and USB ports where a Thunderbolt 4 interface has also been installed.
Please Email us for the complete specifications.