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Walz PAM-2500 Handbook Of Operation

Portable chlorophyll fluorometer
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PAM-2500
Portable
Chlorophyll Fluorometer
Handbook of Operation
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 +49 9133 77650 · Telefax +49 9133 5395
info@walz.com · www.walz.com
Printed in Germany

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  • Page 1 Handbook of Operation 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 +49 9133 77650 · Telefax +49 9133 5395 info@walz.com · www.walz.com Printed in Germany...

  • Page 3: Table Of Contents

    3.3.1 Software Update............... 17 3.4 Accessories ................17 3.4.1 Field Combo (optional) ............. 17 3.4.2 Additional Components ............ 19 4 PAM-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 ..........
  • Page 4 CONTENTS 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 ................
  • Page 5 CONTENTS 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 ....
  • Page 6 CONTENTS 9.2 Conditions ................143 9.3 Instructions ................144 9.4 Applicable law ................ 145 10 Index ..................147...
  • Page 7 CONTENTS...

  • Page 9: Safety Instructions

    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...

  • Page 11: Introduction

    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.

  • Page 12: Intention Of This Handbook

    CHAPTER 2 INTRODUCTION 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”...

  • Page 13: Important Points

    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.

  • Page 15: Components And Setup

    ) 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.
  • Page 16 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)

  • Page 17: Pamwin-3 Software Installation

    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.
  • Page 18 “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).

  • Page 19: Software Update

    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-...
  • Page 20 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).

  • Page 21: Additional Components

    3.4.2.1 Special Fiberoptics 2010-F The Special Fiberoptics 2010-F are connected to the front side of the PAM-2500 control unit with the help of a special plug that resembles an electrical connector. There are three “fiber pins” with different optical cross-sections, which fit into the corresponding holes at the front side of the PAM-2500 housing, where they interface with the various light sources and the photodiode detector.
  • Page 22 CHAPTER 3 COMPONENTS & SETUP leaves, above the hole (compare Fig. 3). In the latter case, the leaf can be kept in place by the folded part of the clip. Fig. 3: Distance Clip (to position leaf with respect to fiberoptics) The distance between the fiberoptics exit plane and sample has con- siderable influence on signal amplitude and effective light intensity (Fig.
  • Page 23 It features special mini-quantum and temperature sensors, the readings of which are transferred to the PAM-2500 with every Satu- ration Pulse measurement. Placed in the holder, the leaf is resting on a Perspex tube with broad-...
  • Page 24 Tripod ST-2101) facilitates long measurements with the same plant. The handle of the Leaf-Clip Holder 2030-B features a red push- button for remote control of the PAM-2500. Pressing this button ini- tiates a Saturation Pulse. Fig. 5: Leaf-Clip Holder 2030-B with Fiberoptics 2010- F.
  • Page 25 When the PAM-2500 internal Actinic Light sources are applied via the fiberoptics (Fig. 5), e.g. for recording of light response curves, the sensor should be switched off via the soft- ware (see Advanced Level, General Settings, Section 4.3.1) and the...
  • Page 26 CHAPTER 3 COMPONENTS & SETUP For the reliability of the calibration it is critical to keep the diffuser clean. It is advisable to check calibration regularly by comparison with a standard quantum sensor. Any deviation can be corrected by entering a recalibration factor in the Options menu of the PamWin-3 program (see Section 4.3.1.1).
  • Page 27 COMPONENTS & SETUP CHAPTER 3 Fig. 6: Fiberoptics holder for Surfaces 2060-A (top) and Arabidopsis Leaf Clip 2060 (bottom) in combination with the mini-quantum/temp. sensor 2060-M.

  • Page 28: Dark Leaf Clip

    CHAPTER 3 COMPONENTS & SETUP 3.4.2.5 Mini-Quantum/Temp.-Sensor 2060-M (optional) The Mini-Quantum/Temp.-Sensor 2060-M essentially has the same features as outlined above for the Leaf-Clip Holder 2030-B, except that the mini-sensors of PAR and temperature are not mounted in a leaf-clip. This device is rather designed for experiments with objects which are not leaf-shaped, like crustose lichens and cushions of moss.
  • Page 29 COMPONENTS & SETUP CHAPTER 3 When the leaf clip shutter is closed and the Measuring Light is on, an artefactual Ft signal is observed. This signal is due to a small fraction of the Measuring Light which is reflected by the closed shutter and reaches the photodetector.
  • Page 30 COMPONENTS & SETUP 3.4.2.7 Suspension Cuvette KS-2500 (optional) The PAM-2500 can also be used for the measurement of suspen- sions. For that purpose, the suspension Cuvette KS-2500 was devel- oped, which can be connected to a water bath to control the sample...

  • Page 31: Pam-2500 Operation

    Field Screen The <Field Screen> of the PamWin-3 software has been developed for outdoor operation of the PAM-2500 fluorometer where ease and simplicity of instrument control is important. Conveniently, the ele- ments of the <Field Screen> are accessible via the display of a touch screen PC.

  • Page 32: Monitoring Graphs

    CHAPTER 4 PAMWIN-3 FIELD SCREEN Fig. 9: Field Screen Overview. 4.2.1 Monitoring Graphs The <Field Screen> provides 2 monitoring graphs (Fig. 9): the left graph records slow fluorescence changes, and also depicts Fm and Fmʹ values as small crosses. Generally, only those Saturation Pulse analyses which result in graphical display of Fm or Fmʹ...
  • Page 33 PAMWIN-3 FIELD SCREEN CHAPTER 4 adapts the Y axis range of both monitoring screens to the signal am- plitude. The actual signal amplitude (in Volts) can be read from the Y-axes. In both screens, the Fo and Fm levels are displayed as yel- low and red dashed lines, respectively (Fig.

  • Page 34: Program And Script Control

    CHAPTER 4 PAMWIN-3 FIELD SCREEN Fig. 11: Field Screen. The blue square surrounds the four fields for the se- lectable parameters. On clicking on one of these four fields a menu pops up that allows the selection of a parameter for this field. 4.2.2 Program and Script Control The Program and Script Control section (Fig.

  • Page 35: Alphanumeric Area

    PAMWIN-3 FIELD SCREEN CHAPTER 4 4.2.3 Alphanumeric Area The elements of the alphanumeric area are divided into four groups: (1) light control, (2) primary fluorescence data, (3) fluorescence ratio parameters (see Fig. 11 for the selection of these parameters), and (4) additional data like PAR and temperature (Fig.
  • Page 36 CHAPTER 4 PAMWIN-3 FIELD SCREEN Group 1 (Light Control, see Fig. 12) includes “status” and “set” but- tons A status button either turns on or off a function (e.g. the Meas- uring Light) or it changes the light color (e.g. red to blue Actinic Light).
  • Page 37 PAMWIN-3 FIELD SCREEN CHAPTER 4 MF-H Clicking on the <MF-H> button switches between low and high Measuring Light frequency. Grey background color indicates low frequency (e.g., for Fo determination) and green background color indicates high frequency (e.g., during actinic illumination). Measuring Light frequencies can be modified at the Advanced Level of WinControl-3.
  • Page 38 ·s), respectively. Level and PAR value are quasi-linearly related. (PAR measured by MQS-B Quantum Sen- sor and a Universal Light Meter (ULM, Walz) at standard distance and 60° optical geometry of a 2030-B Leaf-Clip Holder.) 0.01 s: Using this button permits the setting of the duration of a Saturation Pulse in steps of 10 ms, with 1 = 10 ms and 80 = 800 ms.
  • Page 39 PAMWIN-3 FIELD SCREEN CHAPTER 4 greens, the nick name of cyanobacteria, absorb both blue and green light badly. With red as the Actinic Light source, choosing blue in the <PS I light> section will turn these LEDs into a second Actinic Light source for non-cyanobacterial algae and plants.
  • Page 40 CHAPTER 4 PAMWIN-3 FIELD SCREEN 4.2.3.2 Fluorescence Data Dark-acclimated sample With a dark-acclimated sample, clicking on Fv/Fm records two types of Saturation Pulse data (see “Fluor.Data” in Fig. 12): Fo Minimum chlorophyll fluorescence yield when all photosystem II reaction centers are open, recorded with low Measuring Light in- tensities (i.e.
  • Page 41 PAMWIN-3 FIELD SCREEN CHAPTER 4 Fmʹ Maximal chlorophyll fluorescence yield when photosystem II reaction centers are closed by a strong light pulse. The Fmʹ is low- ered with respect to Fm by non-photochemical quenching. Ft The Ft denotes the continuously recorded fluorescence. The val- ue of Ft measured shortly before a Saturation Pulse with light- exposed samples is denoted “F”.
  • Page 42 CHAPTER 4 PAMWIN-3 FIELD SCREEN calculated data, definitions are provided in Chapter 5: <Definitions and Equations>. Only four of the possible eight parameters can be displayed simulta- neously. Therefore, for each of the 4 display panels (see: ETR, Y(NPQ), Y(NO), Fluoresc. Quenching, Fig. 12), one parameter can be selected from a list which appears after a left-click on a parameter field.
  • Page 43 PAMWIN-3 FIELD SCREEN CHAPTER 4 Y(NO) – Quantum yield of non-photochemical energy dissipation in PS II other than that caused by down-regulation of the light- harvesting function and/or an increase of the rate constant for heat dissipation. YNPQ – Quantum yield of non-photochemical energy dissipation in PS II due to down-regulation of the light-harvesting function and/or an increase of the rate constant for heat dissipation.
  • Page 44 Actinic Light source of the PAM-2500 is used. Volt – Battery voltage. A completely charged battery shows volt- ages up to 13.7 Volts. At Voltages below 10.5 Volts, the PAM-2500 operates unreliably, particularly during application of Saturation Pulses which require high current flows.

  • Page 45: First Measurements Using The Field Screen

    Information on photosynthesis is obtained when the yields of fluo- rescence induced under different illumination conditions are com- pared. For this purpose, the PAM-2500 has various built-in light sources. When you click the Act. Light key, you will see that the leaf is illuminated by relatively strong red light.
  • Page 46 CHAPTER 4 PAMWIN-3 FIELD SCREEN called Z-scheme of photosynthesis and by the theory of fluorescence quenching. Far-red light is preferentially absorbed by PS I, which, in the absence of actinic illumination, pumps electrons out of the inter- system electron chain. In the process, re-oxidized PQ molecules can bind to the Q -site re-opening PS II reaction centers by allowing the –...
  • Page 47 PAMWIN-3 FIELD SCREEN CHAPTER 4 The Y(II)-determination can be repetitively triggered using the Clock key: the Saturation Pulses now will be applied at time inter- vals defined in the <s:>-time field below the Clock key. The fluo- rescence data measured during Saturation Pulse events are automati- cally stored in the Report file, which can be accessed via the Report key.

  • Page 48: Pamwin-3: Advanced Level

    The <General Settings> screen is the start window for the user inter- face of the <Advanced> level. Under <General Settings>, the mode of operation of the PAM-2500 is selected (either <SP-Analysis> or <Fast Acquisition>) and the PAM-2500 settings are adjusted. The icons and display fields to the right of and below the <General Set-...
  • Page 49 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 so, menu items can be accessed by Windows shortcuts: press down and hold the <Alt> key, and sequentially type the first (underlined) letter of the menu title and the first (underlined) letter of the menu item.

  • Page 50: Script File

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL File The file menu includes items for loading and execution of PamWin-3 Script files which are used to automatically perform measuring rou- tines. Also, printer selection, printing and program closure can be carried out using the <File> menu. Window The <Window>...
  • Page 51 Light Calibration Here, a factor can be entered to calibrate a quantum sensor connected to the PAM-2500 (Factor range: 0.2 to 5). For normal operation of the quantum sensor of the Leaf Clip Holder 2030-B, the default fac- tory value of 1.000 applies.

  • Page 52: Par Lists

    (400-700 nm) of many green leaves. AL Current/PAR Lists All light sources of the PAM-2500 are LEDs which show a very re- producible relationship between LED current and light output. Therefore, illumination conditions with PAM-2500 light sources are well-defined for a particular optical geometry between sample and fiberoptics tip (60°...
  • Page 53 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Note that in all default PAR lists, the PAR value at AL setting <0> (Actinic Light off) represents the integrated Measuring Light intensi- ty at ML intensity setting 10 and a 100 000 Hz measuring pulse fre- quency.

  • Page 54: Distance Clip

    The <Service> menu is required for firmware updates (the software residing on instrument processors is called firmware). New firmware is provided by the Walz Company on a regular basis. To determine your firmware version, click on the menu item <Read Firmware Ver- sion>.
  • Page 55 Download HexFile  Program RISC . Finalize TINY pro- gramming by an OK in the <Program RISC> window, the (program restart) information window, and the <PAM-2500 Controller Ser- vice> window. Close and restart the PamWin-3 program. Trigger out with SP A somewhat unexpected item on the Service menu is <Trigger out...

  • Page 56: Measuring Light Frequency

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Help In the Help menu, checking the option Tooltips activates the Pam- Win-3 tooltips function described earlier (Section 4.1). Clicking on Info displays information on the current PamWin-3 version. 4.3.1.2 Measuring Light Frequency Measuring Light frequencies are set in the <General Settings> win- dow (see Fig.
  • Page 57 Four buttons are involved in the PAM-2500 <Settings> management (see Global Settings in Fig. 13). Default...
  • Page 58 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Other Commands Zoff The <Zoff> (Zero Offset) serves to suppress a “background signal” which does not originate from the investigated sample. It is subtracted from all fluorescence signals. Background signals can arise from traces of scattered Measuring Light reaching the photo- detector.
  • Page 59 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 The C/Fo value is a measure for the contribution of non-PS II fluo- rescence to Fo. This is likely constant PS I fluorescence although other fluorescence sources like free LHCII aggregates cannot be ex- cluded. The assessment of C/Fo requires that the Foʹ mode is acti- vated and that non-photochemical fluorescence quenching decreases the Foʹ...
  • Page 60 TR Pulse . When in the <Pulse Widths> window S+H off is checked, the sam- ple-and-hold amplifier of the PAM-2500 is switched off for the dura- tion of the ST flash. The deactivation of the sample-and-hold ampli- fier prevents artifactual signals arising from the extremely high light intensity of the flash.
  • Page 61 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 The following two buttons are used for illumination in the seconds range in combinations with Saturation Pulse analysis. AL + Y – The command illuminates the sample with Actinic Light as defined in <General Settings> and thereafter carries out a Satura- tion Pulse analysis.
  • Page 62 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL 4.3.1.10 Clock Event In total, 6 different events can be repetitively triggered by the clock function. All events are defined under <General Settings> except for the light curve, which is defined in the <Light Curve> window. The- oretically, the interval time between events ranges from 3 to 900 sec- onds.

  • Page 63: Sp-Analysis Mode

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Usually, a <Record> starts with an <Fo, Fm> determination using a dark-adapted sample. The Fo and Fm fluorescence levels play a role in the calculations of many fluorescence ratio parameters (see Chap- ter 5). It is, therefore, important to carry out proper <Fo, Fm> deter- minations and to correctly organize the recording of Saturation Pulse data (i.e.
  • Page 64 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL troduced in the previous section (<General Settings>). The <Menu Bar> of the <Slow Kinetics> and <General Settings> windows are similar. A difference is that the <File> menu contains as additional commands <Print Graph> and <Open pws-file>, i.e., a “PamWin Slow Kinetic”...
  • Page 65 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 colors of the data points and the background colors of the corre- sponding checkbox labels are identical. 4.4.1.5 X-Axis Scaling/Zoom In Clicking on the left arrow (◄, see Fig. 16) doubles the total x axis time range, whereas the total x axis time range is halved, when the right arrow (►) is clicked.

  • Page 66: Light Curve

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL the relaxation of non-photochemical quenching is monitored (Ind.+Rec.). The Start and Stop buttons in the < Slow Kinetics Control> area start and end a <Record>. Several <Records> can be stored in one <Report>. Recordings of Ind.Curv. and Ind.+Rec. are always initiated by au- tomated <Fo, Fm>...
  • Page 67 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 representation of the results is called a (Rapid) Light Curve. For more information on Rapid Light Curves see Section 5.5. Fig. 17: Light Curve Window. In the present version of the software, below the fit button, either the EP (Eilers an Peeters fit model) or the Platt et al. fit model can be selected.

  • Page 68: Light Curves

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL 4.4.2.1 Display Control Light Curves are plotted with PAR (in µmol photons/(m ·s)) as the abscissa parameter. Ordinate data are selected by checking ETR or Fluoresc. in the Display Control field (see Fig. 17): when Fluo- resc.
  • Page 69 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Programmable parameters are the PAM-2500 Actinic Light intensity setting (1 to 20 in the Intens. column) and the time intervals of a light intensity step (in 10 seconds intervals, Time/10s). To change a setting, click in the target field once and enter the new number. The change becomes effective upon the next click.

  • Page 70: Report

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL In the Light Curve Fit window, besides the three cardinal parameters obtained from fitting procedure, also Fv/Fm x ETR factor/2 is displayed. This is meant to remind the user that Fv/Fm corresponds to the maximum quantum yield of charge separation in PS II reaction centers, from which the parameter alpha can be predicted, when the ETR factor (normally 0.84) and the fraction of quanta distributed to PS II (normally 0.5) are considered.

  • Page 71: Report Window

    Report data. The <Clear Report> com- mand also initiates a new Report by writing a PAM-2500 <Settings> line at the top of the new Report: the line of PAM-2500 <Settings> consists of 16 parameters (see Table 3). Similarly, the <Write to Re- port>...
  • Page 72 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Table 3: Report of Instrument Settings Abbreviation Parameter Actinic Light Intensity Actinic Light Width Blue Light Intensity Blue Light Width Damping Far-red Light Intensity Far-red Light Width Gain High Measuring Frequency Slow Induction Delay Slow Induction Clock Slow Induction Actinic Light Width Low Measuring Frequency Measuring Light Intensity...
  • Page 73 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Second and Third Line: Fo, Fm The second and third Record lines contain data related to an <Fo, Fm> determination, which normally should be carried out at start of a new Record. The abbreviations <IC> and <IC+> in the second line indicate that the measuring programs <Induction Curve>...

  • Page 74: Fast Acquisition Mode

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Actinic light intensity (PAR, µmol photons/(m ·s)). Light inten- sity (PAR, µmol photons/(m ·s). Fo (header lines) and F (subsequent data lines) in Volts. Electron transport rate (ETR, µmol electrons/(m ·s)) Maximum (header lines) and effective photochemical quantum yield (subsequent data lines) of PS II Coefficient of photochemical fluorescence quenching (qP).

  • Page 75: Fast Settings

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Fig. 19: Fast Settings Window. 4.5.1 Fast Settings The <Fast Settings> window represents the user interface for pro- gram timing and light conditions of fast kinetics recordings. In this window, the various tools required to design particular fast kinetics trigger programs are grouped in different window areas according to their functionality (Fig.
  • Page 76 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL 4.5.1.1 Basic Settings Read with Start Conditions – Trigger files contain information on instrument settings (On/off status of measuring light, actinic light, far-red light, high measuring light frequency, logarithmically de- creasing measuring pulse frequency: MF log). Further settings saved are low and high frequency of measuring light pulses (MF-L and MF-H).) When “Read with Start Conditions”...

  • Page 77: Trigger Tool Icons

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Target Averages – The “Target Averages” correspond to the number of Fast Kinetics recordings that will be averaged, when Av- eraging is enabled via the <Averaging> checkbox in the <Fast Kinet- ics> window. Consecutive recordings can be either triggered manual- ly (Fast Kin.
  • Page 78 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Eight different events can be triggered (see Trigger Settings ( II ) and Table 4). Among these events, the ST (single turn-over flash) needs to be very short to induce only a single charge separation in all PS II Table 4: Trigger Chart: Checkbox Panel Checkbox Action...
  • Page 79 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 reaction centers. Only the <High Pulse> and <All Low> buttons are available for configuring the ST. All other events can be triggered by pulses and by signal low/high shifts. To set a pulse: a) Select an event by clicking on its checkbox label (Fig. 19. Trigger Setting ( II )).
  • Page 80 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Table 5: Fast Settings - Trigger Program Tools* Active Action Key Combination Parameters Icon M a i n T r i g g e r C h a r t High <Shift> + <right mouse key> All pulse*,** <Shift>...

  • Page 81: Trigger Pulse

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Longer-term triggering is achieved by using the <Toggle Low/High> tool (Fig. 20). Basically, the procedure is identical to the setting of trigger pulses but in step c) the <Toggle Low/High> icon must be se- lected and, in step e), the setting of the <Trig on, µs> time point does not automatically results in an entry in the <Trig off, µs>...
  • Page 82 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL with a drop down list of pretrigger time intervals. Also, a small chart, entitled <Pre>, to the left of the <Trigger Graph> becomes apparent. In the <Pre> chart, a trigger signal can be toggled between low and high by left-clicking on that trigger line.
  • Page 83 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 4.5.1.4 Light & Pulses In the <fast acquisition> mode, two buttons are available which first start an illumination with Actinic or far-red light and then initiate the execution of a fast kinetics (Fig. 19): The AL + F.K. command starts an Actinic illumination and then executes the current trigger pattern.

  • Page 84: Fast Kinetics

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL rows, but the interval can also be entered manually after double-click on the displayed time interval. 4.5.2 Fast Kinetics The data acquired in the <Fast Kinetics> mode are displayed in the <Fast Kinetics> window. Most options available for axis control in the <Slow Kinetics>...
  • Page 85 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Fig. 21: Fast Kinetics Window. For the transient shown in the figure, 5 meas- urements (spaced 4-5 min apart) were averaged. A 50 µs ST was applied after 2 ms of illumination for the determination of the I1-max level.
  • Page 86 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Up/Down arrow pair to the right of the file name display, or the up/down arrows on your keyboard, or the mouse wheel. 4.5.2.3 File Menu <Fast Kinetics> which are not saved will be lost at closure of Pam- Win-3.

  • Page 87: Zero Time Shift

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 affect the original data but at a certain point will start to dampen the graphically observed features. Start – The command starts immediate execution of the current, loaded, trigger pattern. Auto ML On – If checked, the Measuring Light is switched on at the start of a <Fast Kinetics>...
  • Page 88 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL tively recorded <Fast Kinetics> will be calculated. Clicking on “Start” triggers an execution of the loaded script file and leads to a single measurement. The user controls the time interval between measurements. The number specified for <Target Average> in the <Fast Settings>...

  • Page 89: View Mode

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 4.8 the creation of trigger files is discussed a second time using a slightly different approach. View Mode The <View Mode> of PamWin-3 is dedicated to analyses of fluores- cence measurements. Three windows are available in the <View Mode>: the <Report>, <Slow Kinetics>...
  • Page 90 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL spective data in the <Record> data window: only the selected param- eters are exported as CSV data or to the clipboard. Fig. 22: View Mode Window.

  • Page 91: Script Files

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 Script Files Script files are used for automatic execution of experimental proce- dures of various complexities. Carrying out experiments by Script files can be advantages when the same type of analysis needs to be repeated frequently or when complicated protocols must be exactly reproduced.
  • Page 92 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL - A number of <Editing Tools> (Fig. 23) are provided to compose a Script file. Similar as for the <Command Box>, the <Help> as- sociated with the <Editing Tools> comprehensibly explains their function. Fig. 23: Script Window.

  • Page 93: Data Management

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 4.7.1 Data Management Four commands are provided for script file management: New Script File. Clears the script file window and prompts for a new script file name. Open Script File. Opens a script file with name format “file- name.PRG”.
  • Page 94 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL Insert Generally, the two “Insert” icons transfer commands from the command boxes to the script file window. The upper insert icon transfers commands from the box “Program Control Com- mands”, the lower icon transfers commands from the “General Commands”...

  • Page 95: Command Box (List Of Script File Commands)

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 4.7.3 Command Box (List of Script File Commands) The command box of the script file window (see Fig. 23) consists of 3 sections. The upper one contains commands controlling the pro- gress of script files. The middle panel contains the titles of the 8 groups of commands listed in the bottom panel.
  • Page 96 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT End of Repeti- Marks the end of a repetition Number of repetitions tion Block; block. A repetition block Loops = may contain other repetition blocks. Repetitions can also be terminated depending on the levels of Ft or tempera- ture (see section “Condition Commands”...
  • Page 97 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT Paste to Slow Writes the content of the last None Chart Com- comment-generating line ment Line (printed in blue) into the title line of the Slow Kinetics window. The title line is saved in the “Record Com- ment File”...
  • Page 98 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT Comment = Inserts a comment line in the Comment text script file. A comment line can be written to the slow chart title using the “Paste to Slow Chart” command or to the fast chart title using the “Paste to Fast Chart”...
  • Page 99 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT Start Light Starts recording of a light None Curve curve: equivalent to the “Start” button in the light curve window. Stop Light Stops the execution of a None Curve light curve; equivalent to the stop button of Light Curve window.
  • Page 100 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT Open Lcp File Opens standard light curve File name program file optimized for various types of samples. Clock Starts or stops the repetitive Check/uncheck trigger. Clock Time Sets or modifies clock inter- Clock interval in s (=s).
  • Page 101 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT Zoff List for As indicated, this command Checked is on/unchecked all gain set- determines the Zoff lists for is off tings all gain settings. It is an ex- tension of the previous command.
  • Page 102 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT FR+Yield/Fast Performs far-red pre- None Kin. illumination followed by saturation pulse analysis (in SP Analysis Mode) or fol- lowed by fast kinetics (in Fast Kinetics Mode) as de- fined by the loaded trigger file.

  • Page 103: Aux Socket

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT TR Pulse Applies 5 V trigger pulse at None AUX socket. Section 4: -- Measuring Light Commands -- F ML Switches measuring light Check on/off on/off. MF-H Switches measuring light Check on/off frequency between low and high as defined in the set- tings window.
  • Page 104 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT New Fast Kin. Stops averaging of fast ki- None Average netics measurements and saves averaged kinetics. Auto ML on Turns on measuring light Checkbox just before start fast kinetics measurements, and switches measuring light off immedi- ately thereafter.
  • Page 105 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT Section 6: -- Actinic Light Settings -- Sets intensity levels of: AL-Int. Actinic light, 1 to 20 BL-Int. Blue light, 1 to 20 FR-Int. Far-red light, 1 to 20 SP-Int. Saturation pulse, 1 to 20 Fm-SP-int.

  • Page 106: Single Turnover Flash

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT Sets increase/decrease time interval of: ST Pulse Number of increments to Width Step = Single turnover flash (in- decrease (- #) or to in- crement = 5 µs), MT Pulse crease (+ #) Width Step = Multiple turnover pulse (in- crement = 1 ms for width <...
  • Page 107 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT Section 7: -- Measuring Light Settings -- MF-max Sets the maximum frequency 1000-200000 (Drop down for the measuring light list) MF-H = High measuring light fre- 1000-100000 quency (Hz). (Drop down list) MF-L = Low measuring light fre- 10-5000...
  • Page 108 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL PARAMETER COMMAND, COMMENT INPUT Section 8: -- Condition Commands -- If Ft Executes next command if Ft in Volt Ft is greater (>) / smaller (<) than the threshold value en- tered If Temp Executes next command if Temperature temperature is greater (>) / in °C...

  • Page 109: Practical Tips For Creating Trigger Patterns

    PAMWIN-3 ADVANCED LEVEL CHAPTER 4 PARAMETER COMMAND, COMMENT INPUT End of Rep. Terminates a repetition Temperature Block; Repeat block if temperature is in °C until Temp greater (>) / smaller (<) than the threshold value entered Wait until If the clock counter reaches Number clock counter the set value the next com-...

  • Page 110: Writing Scripts

    CHAPTER 4 PAMWIN-3 ADVANCED LEVEL can be turned off at -10 µs. Enter -10 and push enter. Then enter the endpoint of this event, e.g. 80 µs, and press again enter. To move the event to longer times, first change the end point time to for example 100080 µs and press enter (gives an event that starts at -10 µs and ends at 100080 µs) and then change the start point for example to 99990 µs.
  • Page 111 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 ever, if you want to keep a series of measurements together in one record file this command is not a good idea. In that case it is better to start new records manually making use of the New Record but- ton in the bottom panel of all windows.
  • Page 112 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL high time resolution, or in the case where even minor actinic ef- fects of low frequency measuring light are unwanted, it may be better to switch the measuring light only on during a measurement. To make sure that the measuring light is not accidently left on, a command can be included: F ML Off If the user wants to restrict the use of measuring light, the easiest...
  • Page 113 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 mean the signal induced by this ST or MT will be recorded. For that, it is necessary to define this ST or MT in a trigger file. To execute a trigger file in a script takes the following form (for trig- ger file ST.FTM): Open Fast Kin.
  • Page 114 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL However, what is in this case the time interval between these two single turnover flashes? It is not defined in the script. To find out, we have to go to the pulse definition window (click with the right mouse button on either the MT, ST or TR Pulse buttons to open this window).
  • Page 115 PAMWIN-3 ADVANCED LEVEL CHAPTER 4 The next level of complexity is to create a subprogram out of the commands that have to be repeated and Call this subprogram using the “Call” command each time it is needed. The script finally offers a third possibility. It is also possible to put these lines of script between the commands Begin of Repetition Block and End of Repetition Block;...
  • Page 116 CHAPTER 4 PAMWIN-3 ADVANCED LEVEL sample by a water bath, it would be possible to more or less auto- mate such a measurement. With respect to an unstirred sample the user may want to stop the measurement if the cells start to settle and the Ft value falls below a certain threshold.

  • Page 117: Definitions And Equations

    DEFINITIONS & EQUATIONS CHAPTER 5 Definitions and Equations Relative Fluorescence Yields Typically, five different types of fluorescence levels are acquired by Saturation Pulse analyses. Two of these levels need to be established with the dark-acclimated sample. The three remaining levels are re- peatedly measured during subsequent sample treatments (e.g., expo- sure to Actinic Light;...

  • Page 118: Fluorescence Ratio Parameters

    CHAPTER 5 DEFINITIONS & EQUATIONS Fmʹ Maximum fluorescence level during a treatment; is induced by a saturation light pulse which temporarily close all PS II reactions centers. Fmʹ is decreased with respect to Fm by non-photochemical quenching. F corresponds to the momentary fluorescence yield (Ft) of an illuminated sample shortly before application of a Saturation Pulse.
  • Page 119 DEFINITIONS & EQUATIONS CHAPTER 5 may be severely damaged in Calvin cycle activity and still show a high value of Y(II) in weak light. Therefore, photosynthetic perfor- mance should be assessed during steady state illumination at a pho- ton flux density which is somewhat below saturation in a control sample.
  • Page 120 CHAPTER 5 DEFINITIONS & EQUATIONS Table 8: Fluorescence Ratio Parameters. Source Equation − Maximum photochemical quantum yield of PS II (Kitajima and Butler, 1975) − ′ Effective photochemical quantum yield of ′ PS II (Genty et al., 1989) − ′ Coefficient of photochemical fluorescence quenching (Schreiber et al.
  • Page 121 DEFINITIONS & EQUATIONS CHAPTER 5 and q Coefficients of photochemical fluorescence quench- ing. Both parameters estimate the fraction of open PS II reaction centers. The q is based on concept of separated PS II antennae (puddle mod- el) but the q assumes interconnected PS II antennae (lake model) which appears to be the more realistic situation in leaves (cf.

  • Page 122: Constant Fraction Of Fo Fluorescence (C/Fo)

    CHAPTER 5 DEFINITIONS & EQUATIONS Y(NO) and Y(NPQ) Yields of non-photochemical quenching Genty et al. (1996) first presented expressions based on basic fluo- rescence parameters that describe the partitioning of absorbed excita- tion energy in PS II between three fundamental pathways, expressed in terms of the complementary quantum yields of Y(NO) sum of non-regulated heat dissipation and fluorescence...
  • Page 123 DEFINITIONS & EQUATIONS CHAPTER 5 Here, the assessment of <C/Fo> is based on the comparison of the Foʹ calculated according to Oxborough and Baker (1997) (see Sec- tion 5.1.2) with the Fo’ determined during a post-Saturation Pulse illumination with far-red light. Considering that PS I fluorescence contributes a constant quantity, F , to each fluorescence level, the Oxborough-Baker equation can be reformulated:...

  • Page 124: Relative Electron Transfer Rate (Etr)

    CHAPTER 5 DEFINITIONS & EQUATIONS Relative Electron Transfer Rate (ETR) Relative electron transfer rates are calculated according to: ETR = PAR · ETR-Factor · P · Y(II). The basic idea of the ETR equation is to multiply Y(II), the effective photochemical quantum yield of PS II, by an estimate for the photon flux density absorbed by PS II.

  • Page 125: Rapid Light Curves

    DEFINITIONS & EQUATIONS CHAPTER 5 - the value of ETR/4 is not necessarily identical to CO -fixation rate or O -evolution rate; discrepancies e. g. may arise from photorespiratory electron flow, nitrite reduction or electron cy- cling at PS II - fluorescence information primarily originates from the topmost chloroplast layers, while gas exchange integrates over all layers;...
  • Page 126 CHAPTER 5 DEFINITIONS & EQUATIONS tion is expected (hoped for) during the set of light intensities meas- ured. So-called “Rapid Light Curves” (RLC) may want to character- ize a state, or provide information on the ability, or lack thereof, of the photosynthetic apparatus to respond to rapid changes in the light intensity and are not to be confused with classical photosynthetic light response curves in which photosynthetic rates under steady...
  • Page 127 DEFINITIONS & EQUATIONS CHAPTER 5 α ⋅ ⋅ ⋅ ⋅ Fig. 25 illustrates the behavior of the Eilers and Peeters model func- tion for 3 theoretical cases which show identical values of α but, at high light intensities, different degrees of photoinhibition. Curve Parameter α...

  • Page 128: Some Papers Related To Rapid Light Curves

    CHAPTER 5 DEFINITIONS & EQUATIONS 5.5.1 Some Papers related to Rapid Light Curves Fouqueray M, Mouget J-L, Morant-Manceau A, Tremblin AG (2007) Dynamics of short-term acclimation to UV radiation in marine dia- toms. J Photochem Photobiol B: Biology 89: 1-8 Jassby AD, Platt T (1976) Mathematical formulation of the relation- ship between photosynthesis and light for phytoplankton.

  • Page 129: Literature Cited In Chapter 5

    DEFINITIONS & EQUATIONS CHAPTER 5 Serôdio J, Vieira S, Cruz S, Barroso F (2005) Short-term variability in the photosynthetic activity of microphytobenthos as detected by measuring rapid light curves using variable fluorescence. Marine Bi- ol 146: 903-914 White AJ, Critchley C (2005) Rapid light curves: A new fluores- cence method to assess the state of the photosynthetic.
  • Page 130 PAM fluorometry and the Saturation Pulse method. PAM Applica- tion Notes 1: 27-35 (http://www.walz.com/e_journal/pdfs/PAN078007.pdf) Kramer DM, Johnson G., Kiirats O, Edwards GE (2004) New flux parameters for the determination of Q redox state and excitation fluxes.
  • Page 131 DEFINITIONS & EQUATIONS CHAPTER 5 Peterson RB, Oja V, Laisk A (2001) Chlorophyll fluorescence at 680 and 730 nm and leaf photosynthesis. Photosynth Res 70: 185-196 Pfündel E (1998) Estimating the contribution of Photosystem I to to- tal leaf chlorophyll fluorescence. Photosynth Res 56: 185-195 Pfündel EE, Ben Ghozlen N, Meyer S, Zoran G.

  • Page 132: Some Reviews On Chlorophyll Fluorescence

    CHAPTER 6 REVIEWS ON CHL FLUORESCENCE Some Reviews on Chlorophyll Fluores- cence Baker NR (2008) Chlorophyll fluorescence: A probe of photosynthe- sis in vivo. Annu Rev Plant Biol 59: 89-113 Bernhardt K, Trissl H-W (1999) Theories for kinetics and yields of fluorescence and photochemistry: how, if at all, can different models of antenna organization be distinguished experimentally? Biochim Biophys Acta 1409: 125-142...
  • Page 133 REVIEWS ON CHL FLUORESCENCE CHAPTER 6 Krause GH, Weis E (1991) Chlorophyll fluorescence and photosyn- thesis: The basics. Annu Rev Plant Physiol Plant Mol Biol 42: 313- Logan BA, Adams III WW, Demmig-Adams B (2007) Avoiding common pitfalls of chlorophyll fluorescence analysis under field conditions.
  • Page 134 CHAPTER 6 REVIEWS ON CHL FLUORESCENCE...

  • Page 135: Specifications

    (measuring light, red and blue actinic light, and far red light). Saturation Pulse at maximum in- tensity, 30 W Recharging time: approximately 6 hours (with the PAM-2500 turned off) via Battery Charger MINI-PAM/L Operating temperature: -5 to +40 °C...

  • Page 136: Light Sources

    CHAPTER 7 SPECIFICATIONS Dimensions: 23 cm x 10.5 cm x 10.5 cm (L x W x H) aluminum housing Weight: 2.5 kg (including battery) 7.1.2 Light sources Measuring light: Red LEDs, maximum emission at 630 nm, FWHM (full width at half maximum) 20 nm. 1 µs pulses at modulation fre- quencies 10 to 5000 Hz for Fo determinations (200 Hz default), and 1 to 100 kHz during actinic illumination, fast kinetics with 100 or 200 kHz, 20 intensity levels, frequency dependent effective PAR...

  • Page 137: Special Fiberoptics 2010-F

    Output: PAR, high sensitivity range: 0 to 1000 µmol m-2 s-1 (out- put 0 to 2.5 V for each range). Leaf temperature, -20 to +60 °C (0 to 0.8 V). Remote trigger button, signal line connected to ground. Power supply: PAM-2500 leaf clip socket (5 V/4 mA) Cable length: 100 cm...

  • Page 138: Battery Charger Mini-Pam/L

    7.1.7 External Voltage Supply Cable MINI-PAM/AK Design: 90 cm cable with fuse for over-current protection. The cable bypasses the internal battery of the PAM-2500 fluorometer and sup- plies voltage from an external 12 V battery directly to the electronics 7.1.8 System Control and Data Acquisition...

  • Page 139: Carrier Bag

    Operating system: Microsoft Windows 7/8/10. 7.2 Accessories 7.2.1 Touchscreen tablet computer for Field Research 7.2.1.1 Computer box Design: Aluminum compartment, 23.5 cm x 15.0 cm x 4.5 cm (L x W x H), on top of PAM-2500 chassis to keep an touchscreen tablet computer...

  • Page 140: External Battery 000160101314

    °C (0 to 0.8 V). Remote trigger button, signal line connected to ground Power supply: Via PAM-2500: leaf clip socket (5 V/4 mA) Length of power cable: 100 cm Dimensions: 17 cm x 5.7 cm (max.) x 8 cm (max.) (L x W x H)

  • Page 141: Arabidopsis Leaf Clip 2060-B

    Weight: 310 g 7.2.5 Arabidopsis Leaf Clip 2060-B Design: Aluminum clip with 3.2 mm diameter viewing area designed to position small leaves below the fiberoptics of the PAM-2500, pre- pared to accommodate PAR and temperature sensors of the Mini Quantum/Temperature-sensor 2060-M Dimensions: 7.6 cm x 3.0 cm (max.) x 5.2 cm (max.) (L x W x H)

  • Page 142: Suspension Cuvette Ks-2500

    CHAPTER 7 SPECIFICATIONS Length of sensor cables: 30 cm Dimensions: 16 cm x 3 cm x 1.7 cm (L x W x H) Weight: 220 g 7.2.8 Suspension Cuvette KS-2500 Design: Round stainless steel cuvette (7.5 mm wide, 9.0 mm deep) with top window adapter for connecting the Fiberoptics;...

  • Page 143: Trouble Shooting

    C:\PamWin_3\Data_2500 directory. Start instrument and software. - Unable to establish communication between the computer and the PAM-2500. Close PamWin-3 AND switch off the PAM-2500 fluorome- ter. After several seconds, switch PAM-2500 on again, make sure that the green signal LED flashes and start software.

  • Page 144: Battery Replacement

    8.1 Battery replacement Ordering information for the new battery is provided on the Walz website. https://www.walz.com/support/support.html, open “Battery Table”...

  • Page 145: Guarantee

    - Any defects or damage directly or indirectly caused by or result- ing from the use of unauthorized replacement parts and/or service performed by unauthorized personnel. - Any product supplied by the Heinz Walz GmbH, Germany which has been subjected to misuse, abuse, abnormal use, negligence, alteration or accident.

  • Page 146: Instructions

    Heinz Walz GmbH, Germany. - The product must be returned to Heinz Walz GmbH, Germany, within 30 days after Heinz Walz GmbH, Germany has received written notice of the defect. Postage, insurance, and/or shipping costs incurred in returning equipment for guarantee service are at customer expense.

  • Page 147: Applicable Law

    GUARANTEE CHAPTER 9 - Heinz Walz GmbH, Germany is not responsible or liable for missing components or damage to the unit caused by handling during shipping. All claims or damage should be directed to the shipping carrier. 9.4 Applicable law This Guarantee is governed by German law.

  • Page 149: Index

    INDEX CHAPTER 10 10 Index Advanced Level ........48 Fast Acquisition ........74 General Settings ......48 AL + F.K..........83 AL + F.K..........83 FR + F.K..........83 AL+Y command ........61 Graph ............ 84 AUX socket ........... 60 Setting ...........

  • Page 150: Index

    CHAPTER 10 INDEX Fo ............117 Fo calculation (˜Fo) ....... 59 New Script File ........93 Fo fluorescence ......40, 58 NPQ ........... 121 FR + F.K..........83 FR+Y command ........61 Open Script File ........93 Graphing data ........64 PamWin Help ........
  • Page 151 INDEX CHAPTER 10 Sample & Hold ........109 Thermocouple ......... 26, 28 Saturating Pulse Analysis ....117 Trigger Save Script File ........93 Default ..........88 Script File ..........109 Graph ..........81 Script File Commands ......95 Pretrigger ......... 81 Script File Comment ......

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