Walz PAM-2500 User manual

Printed in Germany
2.156/11.2019
Second Edition, November 2019
PAM_2500_06.doc
Heinz Walz GmbH, 2019
Heinz Walz GmbH · Eichenring 6 · 91090 Effeltrich · Germany
Phone [phone] · Telefax [phone]
[email protected] · www.walz.com
PAM-2500
Portable
Chlorophyll Fluorometer
Handbook of Operation

CONTENTS
1
Contents
1Safety Instructions.......................................................................7
1.1 General Safety Instructions......................................................... 7
1.2 Special Safety Instructions..........................................................7
2Introduction..................................................................................9
2.1 Intention of this Handbook .......................................................10
2.2 Important points........................................................................11
3Components and Setup..............................................................13
3.1 Basic System Components........................................................13
3.2 Basic System Setup...................................................................13
3.3 PamWin-3 Software Installation...............................................15
3.3.1 Software Update...............................................................17
3.4 Accessories ...............................................................................17
3.4.1 Field Combo (optional).....................................................17
3.4.2 Additional Components....................................................19
4PAM-2500 Operation.................................................................29
4.1 PamWin-3 Help.........................................................................29
4.2 Field Screen ..............................................................................29
4.2.1 Monitoring Graphs............................................................30
4.2.2 Program and Script Control..............................................32
4.2.3 Alphanumeric Area...........................................................33
4.2.4 First Measurements Using the Field Screen.....................43
4.3 PamWin-3: Advanced Level.....................................................46
4.3.1 General Settings................................................................46
4.4 SP-Analysis Mode.....................................................................61
4.4.1 Slow Kinetics Window.....................................................61

CONTENTS
2
4.4.2 Light Curve....................................................................... 64
4.4.3 Report ............................................................................... 68
4.5 Fast Acquisition Mode.............................................................. 72
4.5.1 Fast Settings...................................................................... 73
4.5.2 Fast Kinetics..................................................................... 82
4.6 View Mode ............................................................................... 87
4.6.1 Icon Bar ............................................................................ 87
4.6.2 Sidebar.............................................................................. 87
4.6.3 Pick Data........................................................................... 87
4.7 Script Files................................................................................ 89
4.7.1 Data Management............................................................. 91
4.7.2 Editing Tools .................................................................... 91
4.7.3 Command Box (List of Script File Commands)............... 93
4.8Practical tips for creating trigger patterns ........................ 107
4.9Writing scripts....................................................................... 108
5Definitions and Equations....................................................... 115
5.1 Relative Fluorescence Yields ................................................. 115
5.1.1 Measurements with dark-acclimated samples................ 115
5.1.2 Measurements with light-exposed (treated) samples...... 115
5.2 Fluorescence Ratio Parameters............................................... 116
5.3 Constant Fraction of Fo Fluorescence (C/Fo) ........................ 120
5.4 Relative Electron Transfer Rate (ETR).................................. 122
5.5 Rapid Light Curves................................................................. 123
5.5.1 Some Papers related to Rapid Light Curves................... 126
5.6 Literature Cited in Chapter 5.................................................. 127
6Some Reviews on Chlorophyll Fluorescence......................... 130
7Specifications............................................................................ 133

CONTENTS
3
7.1 Basic System........................................................................... 133
7.1.1 General Design ...............................................................133
7.1.2 Light sources...................................................................134
7.1.3 Special Fiberoptics 2010-F.............................................135
7.1.4 Distance Clip 60° 2010-A...............................................135
7.1.5 Leaf-Clip Holder 2030-B................................................135
7.1.6 Battery Charger MINI-PAM/L .......................................136
7.1.7 External Voltage Supply Cable MINI-PAM/AK............136
7.1.8 System Control and Data Acquisition ............................136
7.1.9 Carrier Bag......................................................................137
7.1.10 Transport Box 2040-T ....................................................137
7.1.11 Minimum Computer Requirements ................................137
7.2 Accessories .............................................................................137
7.2.1 Touchscreen tablet computer for Field Research...........137
7.2.2 External battery 000160101314......................................138
7.2.3 Automatic charger 000190101099..................................138
7.2.4 Leaf-Clip Holder 2030-B................................................138
7.2.5 Arabidopsis Leaf Clip 2060-B........................................139
7.2.6 Dark Leaf Clip DLC-8....................................................139
7.2.7 Mini Quantum/Temperature-Sensor 2060-M.................139
7.2.8 Suspension Cuvette KS-2500 .........................................140
7.2.9 Magnetic Stirrer MKS-2500...........................................140
7.2.10 Compact Tripod ST-2101A............................................ 140
8Trouble Shooting......................................................................141
8.1 Battery replacement ................................................................142
9Guarantee .................................................................................143
9.1 Manufacturer’s Guarantee ......................................................143

CONTENTS
4
9.2 Conditions............................................................................... 143
9.3 Instructions.............................................................................. 144
9.4 Applicable law........................................................................ 145
10 Index.......................................................................................... 147

CONTENTS
5

SAFETY INSTRUCTIONS CHAPTER 1
7
1Safety Instructions
1.1 General Safety Instructions
a ) Read the safety instructions and the operating instructions
prior to operation of the device.
b ) Pay attention to all the safety warnings.
c ) Keep the device away from water or high moisture areas.
d ) Keep the device away from dust, sand and dirt.
e ) Do not put the device near sources of heat.
f ) Always ensure there is sufficient ventilation.
g ) Connect the device only to the power source indicated in the
operating instructions or on the device. If the device is not in
use, remove the mains plug from the socket
h ) Clean the device only according to the manufacturer’s rec-
ommendations.
i ) Ensure that neither liquids nor other foreign bodies can get
inside the device.
j ) The device should only be repaired by qualified personnel.
1.2 Special Safety Instructions
The PAM-2500 is a highly sensitive instrument which should be only
used for research purposes, as specified in this manual. Follow the
instructions of this manual in order to avoid potential harm to the us-
er and damage to the instrument.
The PAM-2500 can emit very strong light! In order to avoid harm to
your eyes, never look directly at the fiberoptics end, or at open light
ports on the front side of the control unit.

8

INTRODUCTION CHAPTER 2
9
2Introduction
The PAM-2500 Portable Chlorophyll Fluorometer is the follow-up
model of the well-known PAM-2000/2100 instruments which were
introduced in the 1990s as the first portable PAM fluorometers and
since then have been successfully applied worldwide by numerous
scientists. In the development of the PAM-2500, particular care was
taken to maintain all properties appreciated by PAM-2000/2100 us-
ers and, at the same time, to take account of the recent technical pro-
gress.
Essentially, the hardware and optical system are thoroughly modern-
ized. Also, while continuing basic elements of the graphical user in-
terface, instrument operation is based on the newly-developed Pam-
Win-3 software. The program permits operation under Windows op-
erating systems on normal personal computers, but also on touch
screen tablet computers.
Major points of progress of the PAM-2500 with respect to its prede-
cessors are:
-Use of LEDs (light emitting diodes) for all internal light
sources including Saturation Pulses and Actinic Light.
-Blue and Red Actinic internal light sources.
-Single turn-over and multiple turn-over saturating flashes.
-Time resolution down to 10 µs.
-Easily updateable firmware.
-Optional touch-screen tablet computer.

CHAPTER 2 INTRODUCTION
10
2.1 Intention of this Handbook
The Portable Fluorometer PAM-2500 displays a high degree of flex-
ibility in measuring and analyzing fluorescence. This does not mean,
however, that all features of this multifunctional instrument must be
understood before measurements can be started. Actually, due to the
“intelligent” central control of all functions by the special PamWin-3
software, serious operational mistakes harming the instrument are
highly unlikely. Also, at first, there is no need to care about the nu-
merous settings of instrument parameters, because these are pre-
adjusted for standard measurements. Hence, even the inexperienced
user can start measuring with a minimum of background knowledge
and will be gradually guided to deeper understanding and more com-
plex applications.
This handbook tries to cover all of the numerous features and appli-
cations of the PAM-2500 Fluorometer, some of which probably are
not of immediate interest to many users, but probably will become
relevant, as new questions arise on the basis of the results obtained.
The best way to become acquainted with all features of the
PAM-2500 Fluorometer is to read this handbook section by section,
trying out all described functions. On the other hand, in order to get a
quick start, it will suffice to read Section 4.2.

INTRODUCTION CHAPTER 2
11
2.2 Important points
Help:
All program levels of the PamWin-3 software offer online Help-texts
with information on essentially all active user surface elements. To
access the online Help-texts:
- Move mouse cursor on the window element of interest until a
small tag (“Tooltip”) appears.
- A “Tooltip” remains visible for 2.5 seconds. During this inter-
val, pressing the <@> or <F1>key results in the display of the
relevant Help-text.
PAR sensor:
The continuous Reading of PAR sensor data can be switched off on
the <General Settings> window in the Advanced Mode of PamWin-
3. This should not be forgotten when running a Light Curve protocol!
In that case the instrument will use the ambient light intensity for the
light intensity axis of the Light Curve!
Battery:
Volt Battery voltage. A completely charged battery shows voltages
up to 13.7 Volts. At Voltages below 10.5 Volts, the PAM-2500 oper-
ates unreliably, particularly during application of Saturation Pulses
which require high current flows.
-Note: to prevent deep discharge of the battery, the PAM-2500
shuts off when the battery voltage drops below 9.4 Volts.
-Note: at battery voltages below 11.8 Volts, the PAM-2500 can-
not be switched on.

12

COMPONENTS & SETUP CHAPTER 3
13
3Components and Setup
3.1 Basic System Components
The basic system consists of the following components:
a ) PAM-2500 Control Unit
b ) Special Fiberoptics 2010-F
c ) Distance Clip 60° 2010-A
d ) Battery Charger MINI-PAM/L
e ) MINI-PAM/AK cable
f ) Special USB cable PAM-2500/K1
g ) Fluorescence Standard Foil
h ) Spare Fuse
i ) Carrier Bag
j ) Transport Box 2040-T
k ) Software. PamWin-3 System Control and Data Acquisition
3.2 Basic System Setup
Note: Great caution should be exercised to prevent any dirt or for-
eign matterfrom entering the ports for the fiberoptics pins.
To set up the PAM-2500 fluorometer:
-Connect fiberoptics to the control unit using the three-pin opti-
cal connector on the front side of the unit).

CHAPTER 3 COMPONENTS & SETUP
14
Note: Do not force a plug into the wrong socket. Orient each plug so
that the red dot on the plug coincides with the red dot of the
socket. Do not try to disconnect a plug by pulling at the cable.
Disconnect plug by pulling at the rippled metal part of the
plug.
Fig. 1: PAM-2500 Control Unit.
Front. (AL: Red Actinic Light, SP: Saturation Pulse, ML: Measuring
Light, FR: Far-Red light, BL: Blue Actinic Light, Det.: Detector)

COMPONENTS & SETUP CHAPTER 3
15
-Connect the battery charger cable to the <EXT.DC> socket
(see Fig. 1 A), and to line power: thereafter, a green
<CHARGE> LED (Fig. 1 A) indicates that the internal battery
is charged but yellow LED light signifies that the battery is
currently charging. Please note that an external 12 V battery
cannot recharge the internal battery; but it can provide power
(via the MINI-PAM/AK cable) when the internal battery is
empty.
-Connect computer and control unit (USB socket, Fig. 1 A) by
the PAM-2500 special USB cable.
-Switch on PAM-2500 using the toggle switch labeled POWER
ON (Fig. 1 A). Flashing of the <Status LED> indicates that the
system is ready for communication. The LED will give con-
tinuous light once communication with the computer has been
established. Constant light from the <Status LED> in the ab-
sence of computer communication indicates malfunction of the
PAM-2500. Usually, switching power OFF and, after a couple
of seconds, ON again will restore normal operation.
3.3 PamWin-3 Software Installation
The PAM-2500 software works, at the moment, under Win-
dows 7/8/10.
Depending on the type of CD-ROM delivered, start with a)
or b).
a) CD contains only the setup file (e. g. PamWin_setup.exe),
and the PDF copy of the present manual.
Close all programs, double click on the setup file and follow
instructions. The setup routine will create in the C: root di-
rectory the folder PamWin_3 containing the PAM_WIN.exe

CHAPTER 3 COMPONENTS & SETUP
16
program and accessory files. Further, an icon representing a
shortcut to PamWin-3 is placed on the computer desktop,
and a program group called “PamWin-3” is created in the
program list available in the start menu. The PamWin-3 pro-
gram group includes the uninstall command for PamWin-3.
b) Your CD is entitled “Software & Manuals” and contains a
complete collection of Walz Software & Manuals. The CD
in the CD drive automatically starts the default internet
browser of your computer. If automatic browser start fails,
double-click on the file “index.html” in the root directory of
the CD.
The browser loads the Walz homepage from the CD. Click
“All Products” in the title bar of the homepage, select PAM-
2500, MC-PAM or O2-PAM (the same software controls all
three devices).
Close all programs and install PamWin-3 by clicking on
“EXE-File”. Alternatively, you can copy the PamWin-3
software to any storage medium and use the copied file for
installation. During the
installation process, icons
and shortcuts are created
as described above.
c) Connect PAM-2500 to
computer via USB-cable,
switch Power on and with
minimal delay of about 5 s
start the PamWin-3 soft-
ware by clicking on the “PamWin_3” icon. If the fluorometer
is not switched on or connected, PamWin-3 will ask for the
offline mode of operation. To view and analyze data ac-

COMPONENTS & SETUP CHAPTER 3
17
quired by the PAM-2500, select “PAM-2500” on the “Select
Instrument” window.
3.3.1 Software Update.
The PamWin-3 software is continuously improved. There-
fore, check if on the Walz web site a newer version of the
software is available than that delivered with your instru-
ment. PamWin-3 software for the PAM-2500 is available at:
https://www.walz.com/products/chl_p700/pam-
2500/downloads.html
3.4 Accessories
See Chapter 7 for specifications of accessories.
3.4.1 Field Combo (optional)
The add-ons summarized as <Field Combo> convert the
PAM-2500 chlorophyll fluorometer into a highly mobile
field station, equipped with the options that a normal PC of-
fers and allows working for hours independent of line cur-
rent.
3.4.1.1 Tablet-PC
3.4.1.2 External Battery 000160101314

CHAPTER 3 COMPONENTS & SETUP
18
3.4.1.3 Automatic Charger 0001X
To assemble the <Field Combo> proceed as shown in Fig. 2. and de-
scribed subsequently:
-Piece together <External Battery> and rectangular rubber
foam filler (Fig. 2 A) and place in PAM-2500 carrier bag as
shown in (Fig. 2 B and C).
-Slide-in computer box below elastic belt (Fig. 2 C and D).
Note that the computer box is designed to accept an addi-
tional high-capacity battery to increase the time during which
the instrument can be used independently.
-Insert the Tablet Computer (the model used changes rather
frequently; Fig. 2 E).
Fig. 2: PAM-2500 Field Setup