Walz PHYTO-PAM-II User manual

Printed in Germany
Phytoplankton Analyzer
PHYTO-PAM-II
and
Phyto-Win 3
Software V 3
Principles of Operation
2nd Edition: Nov 2016
PhytoPamII_2.doc
Heinz Walz GmbH, 2016
Heinz Walz GmbH Eichenring 6 91090 Effeltrich Germany
Phone [phone] Telefax +49-(0)9133/5395
Email [email protected] Internet www.walz.com

MAY 2002 CONTENTS
1Safety instructions ........................................................................ 1
1.1General safety instructions .......................................................... 1
1.2Special safety instructions........................................................... 2
2PHYTO-PAM-II........................................................................... 3
2.1Compact Version......................................................................... 5
Spherical Micro Quantum Sensor US-SQS/WB.................... 82.1.1
Stirring Device WATER-S (optional)....................................92.1.2
Battery Charger MINI-PAM/L ............................................ 102.1.3
3PhytoWin Software installation and Phyto-PAM-II setup..... 11
3.1Measure PAR Lists.................................................................... 13
4First fluorescence measurements ............................................. 16
4.1Principle of distinguishing between different groups of
phytoplankton............................................................................ 21
5Saturation Pulse Analysis .......................................................... 23
5.1Measurements with Dark-Acclimated Samples ........................ 23
5.2Measurements with Illuminated Samples.................................. 24
5.3Fluorescence Ratio Parameters ................................................. 25
6Features of PhytoWin Software ................................................ 30
6.1Elements for system operation and display of instrument status33
6.2Channels-window...................................................................... 39
Ft, F, Fm’, Fm’-F, Y(II), Fv/Fm and Mean Value display... 396.2.1
Zero Offset and noise N(t) ................................................... 406.2.2
6.3Algae-window ........................................................................... 42
6.4Settings-window........................................................................ 44
6.5Slow Kinetics window .............................................................. 49
6.6Light Curve window.................................................................. 51
Edit Light Curves ................................................................. 536.6.1
Light Curve Fit-parameters .................................................. 566.6.2

CONTENTS
Comments on Light curves .................................................. 626.6.3
6.7Report Window ......................................................................... 64
6.8Reference-window deconvolution and Chlorophyll
determination calibration values ............................................... 68
How to generate Reference Spectra ..................................... 726.8.1
6.9Fluo Spec -window.................................................................... 74
6.10Fast Kinetik - window.......................................................... 75
O-I1Fit window .............................................................. 776.10.1
Parameters and Output of Fo-I1Rise Analysis ............... 816.10.2
6.11VIEW - MODE .................................................................... 82
6.12Main Menu Bar .................................................................... 85
Options: ETR Parameter ................................................. 876.12.1
Al current/PAR list ......................................................... 886.12.2
6.13Script files ............................................................................ 90
Data Management ........................................................... 916.13.1
Editing Tools................................................................... 926.13.2
List of Script File Commands ......................................... 936.13.3
7Technical Specifications........................................................... 101
PHYTO-PAM-II Compact Version ................................... 1017.1.1
System Control and Data Acquisition................................ 1027.1.2
Battery Charger MINI-PAM/L .......................................... 1037.1.3
Spherical Micro Quantum Sensor US-SQS/WB................ 1037.1.4
Transport Box PHYTO-T .................................................. 1047.1.5
Accessory Stirring Device WATER-S............................... 1047.1.6
8Rechargeable battery ............................................................... 105
9References and further reading .............................................. 106
10Index .......................................................................................... 112
11Warranty.................................................................................. 115

MAY 2002 CONTENTS
11.1Conditions .......................................................................... 115
11.2Instructions......................................................................... 116

CHAPTER 1MAY 2002 SAFETY INSTRUCTIONS
1
1Safety instructions
1.1 General safety instructions
1. Read the safety instructions and the operating instructions
first.
2. Pay attention to all the safety warnings.
3. Keep the device away from water or high moisture areas.
4. Keep the device away from dust, sand and dirt.
5. Always ensure there is sufficient ventilation.
6. Do not put the device anywhere near sources of heat.
7. Connect the device only to the power source indicated in the
operating instructions or on the device.
8. Clean the device only according to the manufacturer’s
recommendations.
9. If the device is not in use, remove the mains plug from the
socket.
10. Ensure that no liquids or other foreign bodies can find their
way inside the device.
11. The device should only be repaired by qualified personnel.

CHAPTER 1 SAFETY INSTRUCTIONS
2
1.2 Special safety instructions
1. PHYTO-PAM-II Multiple Excitation Wavelength Phyto-
plankton and Photosynthesis Analyzer is a highly sensitive
research instrument which should be used only for research
purposes, as specified in this manual. Please follow the
instructions of this manual in order to avoid potential harm to
the user and damage to the instrument.
2. PHYTO-PAM-II employs high intensity LED-array light
sources which may cause damage to the eye. Avoid looking
directly into these light sources during continuous illumination
or saturation pulses.
3. Do not cover the ventilation grille rear side of the instrument.

CHAPTER 2 PHYTO-PAM-II
3
2PHYTO-PAM-II
PHYTO-PAM-II represents the progress of established WALZ
fluorometers dedicated to aquatic research like XE-PAM, WATER-
PAM, previous PHYTO-PAM and MULTI-COLOR-PAM. Choice
of several measuring light wavelengths at variable measuring light
settings (intensity and frequency) offered by XE- and 4-wavelengths
PHYTO-PAM are combined with the high time resolution of
MULTI-COLOR-PAM. Thus enabling classical PAM analysis,
deconvolution of phytoplankton and analysis of fast induction
kinetics.
The PHYTO-PAM-II incorporates a multi-color Chip-On-Board
(COB) LED Array featuring 5 measuring light colors and 6 colors for
actinic light illumination. The 440 nm, 480 nm, 540 nm, 590 nm,
625 nm measuring light facilitates online differentiation of 4
different pigment types therefor deconvolution of green algae,
cyanobacteria, diatoms/dinoflagellates and phycoerythrin containing
organisms like cryptophytes. As in the first generation PHYTO-PAM
deconvolution bases on fluorescence excitation reference spectra.
But for the first time these spectra are not instrument specific but
universal. Due to spectral calibration of PHYTO-PAM-II instruments
reference spectra can be shared between users and instruments.
The PHYTO-PAM-II user surface is based on the proven PhytoWin-
software featuring classical PAM analysis like the estimation of the
effective photochemical quantum yield of PS II and the
determination of photochemical and non-photochemical quenching
parameters. For extended applications a special Fast Kinetics mode
of operation is added for measuring the wavelength-dependent O-I1
fluorescence rise kinetics upon onset of pulses of strong actinic light
information. Thus enabling evaluation of the functional absorption
cross-section of PS II, σPSII, determined by light color and the

CHAPTER 2 PHYTO-PAM-II
4
pigment composition of photosynthetic organisms (Klughammer and
Schreiber, 2015)
Two versions of the PHYTO-PAM-II Phytoplankton Analyzer are
available. A PHYTO-PAM-II Compact Version with integrated
emitter-detector unit and a Modular PHYTO-PAM-II Version
featuring different emitter-detector units connectable to a Power-and-
Control-Unit.
Both versions of the PHYTO-PAM-II are operated by PhytoWin-3
software.