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Walz MINI-PAM-II Manual

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MINI-PAM-II
Manual for
Standalone Use
2.163/04.2018
3. Edition, May 2018
MINI_PAM_II_03.docx
 Heinz Walz GmbH, 2018
Heinz Walz GmbH • Eichenring 6 • 91090 Effeltrich • Germany
Phone +49-(0)9133/7765-0 • Telefax +49-(0)9133/5395
E-mail info@walz.com • Internet www.walz.com

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  • Page 1 MINI-PAM-II Manual for Standalone Use 2.163/04.2018 3. Edition, May 2018 MINI_PAM_II_03.docx  Heinz Walz GmbH, 2018 Heinz Walz GmbH • Eichenring 6 • 91090 Effeltrich • Germany Phone +49-(0)9133/7765-0 • Telefax +49-(0)9133/5395 E-mail info@walz.com • Internet www.walz.com...

  • Page 3: Table Of Contents

    Contents Chapter 1 Contents Contents .............. 1 Safety Instructions ..........1 General Safety Instructions ..........1 Special Safety Instructions ..........1 Introduction ............3 Overview ................4 Components and Setup ........7 Extent of Delivery (Basic System) ........ 7 Optoelectronic Unit ............

  • Page 4: Contents

    Chapter 1 Contents Fluorescence Ratio Parameters ........36 Relative Electron Transfer Rate (ETR) ...... 38 Light Curves ............... 39 Some Light Curve References ........42 Some Reviews on Saturation Pulse Analysis ..43 Hints & Troubleshooting ........45 Instrument Settings ............45 Default settings ..............
  • Page 5 Info ..................101 7.3.7.1 MINI-PAM-II Info ............. 102 7.3.7.2 Sensor Info ..............103 7.3.7.3 Firmware Info ..............104 Specifications MINI-PAM-II ......105 Basic System ..............105 8.1.1 Optoelectronic Unit ............105 8.1.2 Fiberoptics MINI-PAM/F ..........107 8.1.3 Power Supply MINI-PAM-II/N ........
  • Page 6 Chapter 1 Contents 8.1.6 Complementary Items ........... 108 8.1.7 Software WinControl-3 ..........108 8.1.8 Transport Case MINI-PAM/T ........109 Accessories ..............109 8.2.1 2035-B Leaf-Clip Holder ..........109 8.2.2 Fiberoptics Adapter 90° 2030-B90 ......110 8.2.3 2054-L External LED Source ........

  • Page 7: Safety Instructions

    The device and its accessories should only be repaired by qualified personnel. Special Safety Instructions The MINI-PAM-II is a highly sensitive instrument which should be only used for research purposes, as specified...
  • Page 8 The MINI-PAM-II can emit very strong light! In order to avoid harm to your eyes, never look directly into the light port of the MINI-PAM-II or its fiberoptics.

  • Page 9: Introduction

    The instrument continues the tradition of the preceding MINI- PAM chlorophyll fluorometer. The major technical advance- ments of the MINI-PAM-II are the consistent use of energy- efficient LEDs, an internal PAR sensor, and stand-alone op- eration by a touchscreen which is well readable under natu- ral light conditions.

  • Page 10: Overview

    Further, a small section is included providing some hints for be- ginners (Chapter 6, page 45). In the field, the MINI-PAM-II is mostly operated in the stand- alone mode by its touchscreen. Chapter 7 (page 49) provides de- tailed instructions on how to use the touchscreen interface in- cluding advices on fluorescence induction and light curve pro- grams.
  • Page 11 Chapter 3 Also, this manual includes a section on safe handling of the MINI-PAM-II (Chapter 2, page 1), and on the extent of delivery of the basic fluorescence system and its accessories (Chapter 1, page 7). Further, technical information (Chapter 8, page 105)

  • Page 13: Components And Setup

    Components and Setup Chapter 4 Components and Setup Extent of Delivery (Basic System) Optoelectronic Unit MINI-PAM-II/B or -/R Fiber optics MINI-PAM/F Power Supply MINI-PAM-II/N Battery charger 000190101101 Battery (12 x) 000160101990 USB cable type A to Mini B 000130606252 Distance Clip 60°...

  • Page 14: Optoelectronic Unit

    Components and Setup Optoelectronic Unit Sockets, fiber optics port and on/off switch of the back side of the MINI-PAM-II are outlined and numbered in Fig. 1, their properties and functions are explained in Table 1, page 9. Fig. 1: Back panel of MINI-PAM-II Power-and-Control-Unit...
  • Page 15 Port for Fiberoptics MINI-PAM/F Note Great caution should be exercised to prevent dirt or foreign matter from entering the ports or sockets of the MINI-PAM-II. Do not force a plug into the wrong socket. Orientate each plug so that the red dot on the plug coin- cides with the red dot of the socket.

  • Page 16: Emission Spectra

    LED in the MINI-PAM-II/R fluorometer. This LED is electronically driven to act as measuring and as actinic light source. In addition, both versions of the MINI-PAM-II offer a far red LED. Normalized spectra of blue, red and far red LEDs are shown in Fig.

  • Page 17: Batteries

    Components and Setup Chapter 4 4.2.1 Batteries When operated independently, the system is powered by six AA (Mignon) rechargeable batteries (1.2 V/2 Ah). The system can also be powered by non-rechargeable batteries. The battery compartment of the opto-electronic unit does not have a charging function.
  • Page 18 Chapter 4 Components and Setup hole. In the latter case, the leaf can be held between the folded parts of the clip. The distance between fiberoptics exit plane and sample has considerable influence on signal amplitude and effective light in- tensities (Fig.

  • Page 19: Accessories

    MINI-PAM-II is operated by the Win- Control-3 software. The Leaf-Clip Holder 2035-B is almost indispensable for field in- vestigations, when ambient conditions may vary considerably.
  • Page 20 Tripod ST-2101A) facilitates long term measurements with the same plant. The handle of the Leaf-Clip Holder 2035-B features a red push- button for remote control of the MINI-PAM-II. Pressing the button triggers a saturation pulse and associated measurements of fluo- rescence levels for “fluorescence quenching analysis”.
  • Page 21 Components and Setup Chapter 4 Essential optoelectronic elements of this micro-quantum-sensor are: A 3 mm ∅ diffusing disk. High stability silicon photovoltaic detector with filter set for PAR correction, magnetically attached to 2035-B Leaf Clip Holder. Cosine response characteristics (Angular dependence: Error <...

  • Page 22: Dlc-8 Dark Leaf Clip

    LED action Status Flashing green Normal operation. Continuous green Communication from MINI-PAM-II to 2035-B clip interrupted. This happens temporarily during firmware update of MINI- PAM-2. Flashing red (a) Broken thermocouple: inspect and ask for repair kit. (b) Internal error on 2035-B EPROM: contact Walz.
  • Page 23 Chapter 4 Fig. 6: DLC-8 Dark Leaf Clip The tip of the fiberoptics of the MINI-PAM-II fits snugly into the DLC-8 port. With the fiber tip inserted, the sliding shutter of the DLC-8 can be opened so that F and F level fluorescence can be measured without interference of ambient light.

  • Page 24: 2060-B Arabidopsis Leaf Clip

    2060-B Arabidopsis Leaf Clip This leaf clip is designed to position small samples in the beam of the fiberoptics of the MINI-PAM-II. Usually, the 2060-B clip is combined with the 2065-M Mini Quantum/Temp.-Sensor to measure PAR at sample level and lower leaf temperature (see Fig.

  • Page 25: 2065-M Mini Quantum/Temp.-Sensor

    Components and Setup Chapter 4 Fig. 8: 2060-A Fiberoptics Holder for Surfaces & 2065-M Mini Quan- tum/Temp.-Sensor 4.4.5 2065-M Mini Quantum/Temp.-Sensor The light and temperature sensors of the 2065-M device can be mounted on the 2060-B Arabidopsis Leaf Clip and the 2060-A Fiberoptics Holder for Surfaces.

  • Page 26: 2054-L External Led Source

    LED RGBW sources each capable of emitting red, green, blue and white light. Total intensity and color composition can be reg- ulated by the software of the MINI-PAM-II or by WinControl-3. The maximum PAR of each light quality is 1500 µmol m Note Switch off MINI-PAM-II before connecting of disconnection 2054-L External LED Source.
  • Page 27 Components and Setup Chapter 4 Fig. 10: 2054-L External LED Source Fig. 11: Typical emission spectra normalized to unity of red, green, blue and white light of the 2054-L External LED Source...

  • Page 28: Mini-Spec/Mp Miniature Spectrometer

    The spectrometer is calibrated to measure spectra of quantum fluxes. Integration of these spectra over the visible range (400 – 700 nm) yields PAR data equivalent to those recorded by Walz quantum sensors. Like the PAR sensor of the 2035-B or 2065-M devices (Section 4.4.1, page 13 and Section 4.4.4, page 18), the spectrometer...

  • Page 29: Fig. 13 Par Calibration Block

    Chapter 4 can be employed to calibrate the internal PAR sensor of the MINI-PAM-II (see Section 7.3.4.1, page 86). To this aim, the MINI-PAM-II light guide and the entrance optics of the spectrom- eter are inserted in the PAR calibration block (Fig. 13, page 23).
  • Page 30 Chapter 4 Components and Setup Fig. 14: Miniature Spectrometer MINI- SPEC/MP: Configuration A: Configurations of the Miniature Spectrometer. B: Proper alignment of parts for spectrometer configuration using marker triangles...

  • Page 31: Fig. 15 Mini-Spec/Mp

    Components and Setup Chapter 4 In the fluorescence mode, a light guide in the cap leads light from a blue or a green LED located inside the spectrometer body to the sample. The excited fluorescence reaches the spectrometer body through a central hole in the cap. Green light penetrates deeper into photosynthetic tissue than blue or red (Terashima et al (2009) Plant Cell Physiol 50:684-697).
  • Page 32 Chapter 4 Components and Setup viding a sample spectrum (r ) by the spectrum of a white sample light-diffusing reflector consisting of a fluoropolymer with very ( �� ) �� high diffuse reflectance in the entire visible range (r reference ��...

  • Page 33: Ks-2500 Suspension Cuvette

    (FSO2-1) and an interface (FSO2-AK). The sensor is inserted through the needle port of the KS-2500 cuvette (Fig. 17, p 27). The oximeter controls the sensor and transmits the oxygen sig- nal via the interface to the MINI-PAM-II. A separate manual is provided for this package.

  • Page 34: Bcs-9590 Barcode Scanner

    Then, the BCS-9590 Barcode Scanner writes your sample ID into the memory of the MINI-PAM-II or the report data in WinControl-3. For numbers up to 30 000, barcodes are printed in the Aux1 col- umn of the data line of the subsequent saturation pulse data.

  • Page 35: Mini-Pam/F1 Miniature Fiberoptics

    Components and Setup Chapter 4 Fig. 19: BCS-9590 Barcode Scanner 4.4.11 MINI-PAM/F1 Miniature Fiberoptics The MINI-PAM/F1 fiberoptics is useful for investigation of small areas. It consists of a single coated plastic fiber which provides an active diameter of 2 mm. Fig.

  • Page 36: Mqs-B/A Adapter Set For Thin Fiberoptics

    The MQS-B/A Adapter Set is designed to measure the light in- tensity of the MINI-PAM/F1 fiberoptics using an MQS-B light sensor together with a data logger having a high-impedance BNC input, for example, the Walz ULM-500 Universal Light Me- ter & Data Logger. Fig. 21:...

  • Page 37: Saturation Pulse Analysis

    Saturation Pulse Analysis Chapter 5 Saturation Pulse Analysis Five Fluorescence Levels Usually, five different levels of modulated fluorescence are ac- quired by PAM fluorometers. Two of these levels (F and F must be measured with the dark-acclimated sample. The three other levels (F ’, F, and F ’) are measured with the actinic light-...

  • Page 38: Measurements With Dark-Acclimated Samples

    Chapter 5 Saturation Pulse Analysis in both the dark-acclimated and light-exposed state (Table 3, page 34). Because PAM fluorescence is excited by µs pulses of constant amplitude, variations between fluorescence levels are usually in- terpreted as variation in chlorophyll fluorescence yield. This ap- plies for variations between different types of fluorescence levels (e.g.
  • Page 39 Saturation Pulse Analysis Chapter 5 page 31, time point 75 s). Ideally, far red opens PS II in less than 5 s and it is often assumed that during this short interval light-driven energization of the photosyn- thetic membrane decays very little. If the F ’...
  • Page 40 * Kramer et al. (2004) have derived more complex equations for Y(NO) and Y(NPQ). Klughammer and Schreiber (2008) have demonstrated that the equations by Kramer et al. (2004) can be transformed into the simple equations of (Genty et al. 1996) which are used by the MINI-PAM-II and WinControl-3.
  • Page 41 Klughammer C and Schreiber U (2008) Complementary PS II quantum yields calculated from simple fluorescence parameters measured by PAM fluorometry and the Saturation Pulse method. PAM Application 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 42: Fluorescence Ratio Parameters

    Table 3 (page 34) compiles the fluores- cence ratio parameters available in MINI-PAM-II and WinControl- 3. Below, these parameters will be explained briefly. and Y(II) Maximum and effective photochemical quan-...
  • Page 43 Saturation Pulse Analysis Chapter 5 and q Coefficients of photochemical fluorescence quench- Both parameters estimate the fraction of open PS II reaction cen- ters. The q is based on the concept of separated PS II antenna units (puddle model), whereas the q assumes interconnected PS II antenna units (lake model) which appears to be the more realistic situation in leaves (cf.

  • Page 44: Relative Electron Transfer Rate (Etr)

    Chapter 5 Saturation Pulse Analysis Y(NPQ) regulated energy losses of excitation energy by heat dissipation involving ∆pH- and zeaxanthin-dependent mechanisms, and Y(II) use of excitation energy for charge separation. This concept of "complementary PS II quantum yields" is useful to analyze the partitioning of absorbed light energy in photosyn- thetic organisms.

  • Page 45: Light Curves

    Saturation Pulse Analysis Chapter 5 sorbed. The ETR-Factor can be lower in bleached leaves or leaves containing considerable amounts of non-photosynthetic pigments like anthocyanins. (3) P Relative distribution of absorbed PAR to photo- PS1+2 system II The default P is 0.5 which assumes the PS II contrib- PS1+2 utes 50% to total sample absorptance.
  • Page 46 Chapter 5 Saturation Pulse Analysis Fig. 23: Model Functions of Rapid Light Curves. In both cases, RLC and long-term light curves, plotting ETR ver- sus PAR yields light response curves which are often described by the following three cardinal points. α...
  • Page 47 Saturation Pulse Analysis Chapter 5 , µmol electrons/(m · s): Maximum electron • transport rate. , µmol photons/(m · s): Minimum saturating irradiance. • Evaluation of cardinal points requires WinControl-3. The software uses two empirical functions to estimate these cardinal data: the functions REG1 and REG2 have been introduced by Platt et al.

  • Page 48: Some Light Curve References

    Chapter 5 Saturation Pulse Analysis REG2 The function REG2 is monotonically nondecreasing and, hence, does not allow for photoinhibition: α ⋅ PPFD ⋅ tαnh( Here, the cardinal points, α and ETR , are estimated by the fit- ting procedure. With the latter two parameters, the I is calculat- ed as described for REG1.

  • Page 49: Some Reviews On Saturation Pulse Analysis

    Saturation Pulse Analysis Chapter 5 Rascher U, Liebig M, Lüttge U (2000) Evaluation of instant light- response curves of chlorophyll fluorescence parameters ob- tained with a portable chlorophyll fluorometer on site in the field. Plant Cell Environ 23: 1397-1405 Schreiber U, Gademann R, Ralph PJ, Larkum AWD (1997) As- sessment of photosynthetic performance of Prochloron in Lisso- clinum patella in hospite by chlorophyll fluorescence measure- ments.
  • Page 50 Chapter 5 Saturation Pulse Analysis Hanelt D (2018) Photosynthesis assessed by chlorophyll fluores- cence. D-P Häder, GS Erzinger (eds). In: Bioassays. Advanced Methods and Applications, Elsevier, Amsterdam, Netherlands, pp. 169-198 Kalaji HM et al. (2014) Frequently asked questions about in vivo chlorophyll fluorescence: practical issues.

  • Page 51: Hints & Troubleshooting

    Instrument Settings Instrument settings were adjusted at the factory for optimum per- formance of the MINI-PAM-II. For example, LED currents have been adjusted to meet target PAR values for the measuring set- up of the 60° distance clip or the 2035-B leaf clip (Fig. 3 and Fig.

  • Page 52: F Fluorescence

    Chapter 6 Hints Fluorescence Usually, measuring light intensity is adjusted to reach F fluores- cence levels around 500 mV (for a definition of F see Section 5.1.1, page 32). Theoretically, the F should stay below 640 mV. The latter upper value is derived from the assumption that the maximum F of any sample is 0.84 und from the fact that sig- nal saturation occurs at 4000 mV (see equation below, Table 5).
  • Page 53 Hints Chapter 6 Fluorescence The F and F ’ levels are determined as the maximum of the fluorescence signal induced by a saturation pulse. Factory set- tings of saturation pulse width and intensity are adjusted to arrive at a plateau with normal green leaves (Fig. 24A). Some samples (for instance, high light grown leaves) do not reach a plateau with standards settings (Fig.

  • Page 54: Signal Noise

    Chapter 6 Hints Signal Noise Check if fluctuating light sources (fluorescent tubes, computer screens) affect the signal. Exclude that automatic scaling of Y axis at low Y(II) has amplified signal noise. System Hangs Disconnect line power and remove batteries. Power on again. External PAR Sensor is not Responding Check the following: External sensor activated (Fig.

  • Page 55: Touchscreen Operation

    Fig. 25: Touch Screen Calibration Calibration The MINI-PAM-II can be operated autonomously using its resis- tive touchscreen. The touchscreen is factory-calibrated to match its two dimensional response with the LED screen underneath. If touch operation is imprecise, calibration can be performed as...

  • Page 56: Top Level Windows

    Chapter 7 Touchscreen Operation Top Level Windows The MINI-PAM-II provides various windows for control and data display. Frequently used commands, fluorescence data and the actinic light list (PAR list) are presented in 10 top level windows (Table 6, page 50).

  • Page 57: Fig. 26 Principal Screen Layout

    Touchscreen Operation Chapter 7 Generally, all top level windows consist of the “main panel” and the “side panel” (Fig. 26, page 51). The top line of the main panel displays window title and a capital letter which is added as a mark to each saturation pulse analysis.

  • Page 58: Basic Data

    Chapter 7 Touchscreen Operation main menu. Control keys are different between windows. For in- stance, the uppermost button in the sidebar triggers saturation pulse analyses in case of the first three top level windows (cf. Table 6, page 50) but the same button starts automated experi- ments in windows Actinic + Yield, Induction Curve, Light Curve and Recovery Curve.
  • Page 59 Touchscreen Operation Chapter 7 Side Panel Basic Data SAT, trigger saturating pulse to determine Y(II).  , go to next window PAR (int) (Primary Data).  0.138 MENU, go to Main Menu. Y(II) Act.L., actinic light switch. 19.9 MENU Rec Fo,Fm Clock Mark Act.L.
  • Page 60 Chapter 7 Touchscreen Operation The side panel of the Basic Data window offers four commands: the SAT key determines effective PS II photochemical yield, Act.L. represents a switch for actinic light. The MENU key leads to the Main Menu window as in all other top level windows.

  • Page 61: Primary Data

    Touchscreen Operation Chapter 7 7.2.2 Primary Data The insert in the Primary Data window display the fluorescence trace during and right after the last saturation pulse. In addition, the window repeats the data of the previous one (Ft, F Y(II), ETR, PAR). New information of the Primary Data window is the F and F signal levels (after Fo,Fm was pressed) or the F...

  • Page 62: Quenching Analysis

    Touchscreen Operation temperature, °C (Temp) and relative humidity, % (Hum) as measured by the MINI-PAM-II leaf clip (2035-B) are displayed. Also, data from an optional optode oxygen sensor (O2) and two more auxiliary sensors (Aux1 and Aux2) can be displayed.

  • Page 63: Ft-Chart

    The Ft-Chart displays a 25 or 125 s interval of Ft where the rightmost level of the graph corresponds to the current Ft value. The X axis interval can be adjusted in the menu “MINI-PAM-II Settings (Fig. 62, page 96).

  • Page 64: Spectrometer

    Chapter 7 Touchscreen Operation 7.2.5 Spectrometer The SPEC command of the Spectrometer window triggers reading of data from the spectrometer. In the standard mode, the spectrometer is equipped with an optical entrance for external radiation (Fig. 14, page 24). Spectra of photon flux densities are measured in units of nmol photons m .

  • Page 65: Fig. 31Spectrometer

    Touchscreen Operation Chapter 7 white reference material (Fig. 15, page 25), and finally the sam- ple. In the Spectrometer window, touching the graph display area Side Panel Spectrometer | PAR Same as window Quenching Analysis but Act.L. replaced  by SPEC . ...

  • Page 66: Actinic + Yield

    Chapter 7 Touchscreen Operation show a vertical cursor line as well as the x-y data of the intersec- tion between cursor line and spectrum. Use the arrow keys to navigate through the spectra. Touching the graph display again returns to the original function of arrow keys. 7.2.6 Actinic + Yield The “Actinic + Yield”...

  • Page 67: Induction Curve

    Touchscreen Operation Chapter 7 The routine illuminates a sample with actinic light of a defined period. Depending on settings, saturation pulse analysis is car- ried out prior and after actinic light exposure, or only after actinic light exposure. Because, the Actinic + Yield experiment is rather short, it is frequently employed when high light response of pho- tosynthesis needs to be evaluated for many samples.

  • Page 68: Fig. 22 Saturation Pulse Analysis

    Chapter 7 Touchscreen Operation Side Panel Induction Curve START START/STOP. Switch to start or stop fluorescence induction  curve.   .Go to previous, next  window. MENU. Go to Main Menu. MEM. Open “Induction Curve MENU Memory”, view saved induction Fm, F curves.
  • Page 69 Touchscreen Operation Chapter 7 Side Panel Light Curve START START/STOP. Switch to start or stop light curve.    .Go to previous, next window.  MENU. Go to Main Menu. MEM. Open “Light Curve Memory”, view saved light MENU curves.

  • Page 70: Light Curve

    Chapter 7 Touchscreen Operation 7.2.8 Light Curve In analogy to the previous window (Induction Curve), the “Light Curve” window provides buttons to start and stop light curves and to survey them (Fig. 34, page 63). Also, the Light Curve win- dow provides numerical data of saturation pulse analysis which can be accessed by touching the graphics panel.

  • Page 71: Fig. 35 Recovery

    Touchscreen Operation Chapter 7 interval between neighboring saturation pulse analysis roughly doubles with time (Table 7, page 9). Side Panel Recovery A START START/STOP. Switch to start or stop recovery curve.    .Go to previous, next window.  MENU.

  • Page 72: Actinic Light List

    In the factory, the internal actinic light is adjusted so that the PAR values of the Actinic Light List apply to the sample level of the 2035-B leaf clip with the MINI-PAM-II fiber optics fully inserted (i.e. distance between fiber optics tip and sample level of 7 mm;...
  • Page 73 Touchscreen Operation Chapter 7 External versus Internal Actinic Light The main panel of the window Actinic Light List (Fig. 36, page 67) indicates the light source selected for actinic illumination. External Actinic Illumination is displayed when: An external lamp (2054-L External LED Source) is connected to the SYNC port of the MINI-PAM and “PAM Mode (Act)”...
  • Page 74 PAR sensor for various measur- ing configurations. If the calibrated PAR sensor feeds its data in the MINI-PAM-II (as is the case with the PAR sensor of 2035-B leaf clip), the internal PAR sensor can be calibrated automatically. To do so, select “Calibrate”...
  • Page 75 Touchscreen Operation Chapter 7 If “External Actinic Illumination” is displayed by the window “Actinic Light List”, the CAL command adjusts the PAR emitted by the 2054-L External LED Source as measured by an external sensor (typically that of the 2035-B leaf clip) to the PAR values of the Actinic Light List.

  • Page 76: Main Menu

    The “Main Menu” (Fig. 37, page 70) forms the central hub to ac- cess settings, calibration data, hardware information and the memory of the MINI-PAM-II. The Main Menu consists of seven items. Items are selected using the arrow keys of the side panel followed by the SET command, or by directly touching a menu line.

  • Page 77: Meas. Light

    Touchscreen Operation Chapter 7 7.3.1.1 Meas. Light On/Off switch for measuring light (weak excitation light consisting of µs pulses). Measuring light can be switched by selecting “Meas. Light” and touching SET . Independent of the selected line in the menu, the OFF/ON key turns measuring light off or Fig.

  • Page 78: Meas. Light Sett

    Chapter 7 Touchscreen Operation 7.3.1.2 Meas. Light Sett. This command opens the menu Table 8: Measuring Light Fre- quencies “Measuring Light Settings” in which measuring light intensity (Meas. Setting Frequency, Hz Light Int.) can be marked by the Up/Down arrows, selected the SET key and then adjusted us- ing the arrow keys again.
  • Page 79 Touchscreen Operation Chapter 7 The highest measuring light frequency of 100 Hz can be activat- ed by selecting by the SET key the third line in the menu (ML-F High). The ML-F High command overrules settings made under “Meas. Light. Freq.” High measuring light frequency improves signal quality but bears the risk that its higher intensity drives photosynthesis, that is, the measuring light becomes actinic.

  • Page 80: Gain

    The average PAR of measuring light at highest frequency and highest intensity setting was measured to be 1.5 µmol m the PAR sensor of the 2035-B leaf clip and MINI-PAM-II fiber in the fully inserted position. For the same geometrical arrange-...

  • Page 81: Etr-Factor

    Touchscreen Operation Chapter 7 7.3.1.5 ETR-Factor This factor is used for ETR calculations and corresponds to the fraction of incident PAR absorbed by a leaf; its default value is 0.84 (cf. Section 5.3, page 38). 7.3.1.6 Fo’ Mode The “Fo’ Mode” replaces after saturation pulses actinic by far red light to quickly open PS II reaction centers (cf.
  • Page 82 Chapter 7 Touchscreen Operation offset is displayed in the bottom line of the PAM Settings window (Fig. 39, page 72). Procedure Choose dim environment. Switch off any flickering light sources like fluorescent lamps or computer screens. Point fiber tip away from any objects, keep fiber tip clear. Run “Adjust F-Offset”...

  • Page 83: Light Sources

    Touchscreen Operation Chapter 7 7.3.2 Light Sources Actinic Light On/off switch for actinic illumination. Actinic Intensity Intensity regulation actinic light. Select menu item by SET and choose setting using arrow keys. Settings 1 to 12 are available. PAR information of settings is shown in window “Actinic Light List”...

  • Page 84: Far Red Sett

    Chapter 7 Touchscreen Operation Far Red On/off switch for far red light. 7.3.2.1 Far Red Sett. Opens menu to adjust intensity and duration of far red illumina- tion. These settings are active in F ’ determinations 7.3.2.2 Light Panel Sett. Use SET to enter submenu “Light Panel Settings”...

  • Page 85: Sat Settings

    Touchscreen Operation Chapter 7 Note: Operation of the external light source (2054-L External LED Source) requires line power. Note: In absence of an external light source, “PAM Mode Act.” is not available. With “PAM Mode (Act.)”, the total intensity of all four LED groups is determined by the setting for actinic light intensity (see above).

  • Page 86: Program/Clock Settings

    Chapter 7 Touchscreen Operation tance fiber optics tip to sample level: 7 mm; angle between end piece of fiber optics and sample level: 60°). Saturation pulse in- tensity can be adjusted at increments of 500 µmol m 7.3.3 Program/Clock Settings The menu “Program/Clock Settings (Fig.

  • Page 87: Actinic + Yield

    Touchscreen Operation Chapter 7 terval” (up/down keys and SET , respectively) and adjusting time interval (up/down keys). Clock Item Saturation pulse analyses and the programs Actinic + Yield, In- duction Curve and Light Curve can be repetitively performed un- der clock control. Also, recovery experiments can be performed after induction and light curves (item Light Curve + R and Induc- tion Curve + R, respectively).

  • Page 88: Induction Curve

    Chapter 7 Touchscreen Operation actinic illumination without preceding saturation pulse analysis (Initial pulse). Width of actinic illumination is adjusted as de- scribed above for clock interval and initial pulse is selected by the SET command. Further, actinic light intensity is adjusted in the window “Light Sources”...

  • Page 89: Light Curve

    Touchscreen Operation Chapter 7 Width (range 5 s to 10 min) is the time interval between two suc- cessive saturation pulse analyses during illumination. Length is the number of saturation pulse analyses carried out during actinic illumination. Thus, the duration of actinic illumina- tion is “Length –...
  • Page 90 Chapter 7 Touchscreen Operation 2, 3, 4, 5, and 6 will be performed. The time required for a light curve results from Length times Width. Light Curve Settings Side Panel START START. Start light curve Width (mm:ss) 1:00 experiment and switch to Initial Intensity ...

  • Page 91: Sensors

    “Sensor Settings” (Fig. 48). This window consists of five links leading to submenus (distinguished by ) and the option to se- lect for light measurements the MINI-PAM-II internal PAR sensor or the external sensor of the 2035-B leaf clip (Use Ext. PAR/Use Int.

  • Page 92: Internal Par

    (Fig. page 23). Spec. Cal. Factor is correct if both setups result in similar Cali- bration Factors. Ask Walz for the Spec. Cal. Factor when a 2035- B clip is not at hand. 7.3.4.1 Internal PAR “Internal PAR” opens the menu “Int. PAR Sensor Settings” (Fig.

  • Page 93: External Par

    Touchscreen Operation Chapter 7 sor is positioned at a defined distance and angle relative to the end of the fiber optics. Therefore, the readout of the internal PAR sensor applies only for the position of the external PAR sensor. With the external PAR sensor in calibration position, proper cali- bration of the internal sensor is apparent from similar readouts of internal and external PAR.

  • Page 94: Leaf Clip/Ext. Par

    PAR sensor compatible with MNI-PAM-II: To calibrate the inter- nal PAR sensor using the PAR sensor of the 2035-B clip con- nected to the MINI-PAM-II, go to “Calibrate” and establish cali- bration factor automatically. Other PAR sensor: To calibrate the internal PAR sensor using an external PAR sensor which cannot be read by the MINI-PAM-II, adjust calibration factor manually by selecting “Calibration Fac-...
  • Page 95 Touchscreen Operation Chapter 7 sensor connected to the LED-Panel RGBW-L084. In the normal configuration of the MINI-PAM-II, Leaf Clip Ch 1 is selected. External PAR Sensor Side Panel Leaf Clip Ch1   . Up/down Leaf Clip Ch2 keys move cursor.
  • Page 96 Chapter 7 Touchscreen Operation sensor. By considering the spectral variations in sensitivity of the PAR sensor, these calibration values are optimized to measure various light qualities. Specifically, Calib. Sunlight is optimized to measure sunlight under clear skies. Calib. Red, Calib. Green, Blue and Calib.
  • Page 97 Touchscreen Operation Chapter 7 Leaf Clip Ch. 2 in window “Leaf Clip/Ext. PAR Sett” (Fig. 55, page 91) opens the calibration menu for a second PAR sensor connect to the SMA socket on the side of the 2035-B leaf clip. This menu simply consists of slope (Calib,) and offset of the cali- bration line are given.

  • Page 98: Oxygen Sensor

    Oxygen Sensor The window “Oxygen Sensor Settings” is prepared for connec- tion of an optode sensor (oxygen package for MINI-PAM-II) where “Meas. Interval” defines the frequency of oxygen readings and “Blank Out LED” switches of M-PAM-II internal light in the case that it interferes with oxygen measurements.

  • Page 99: Spectrometer

    “Output Format” opens a menu for selection of the parameter used for oxygen represen- tation in the chart. For details see the instructions for the oxygen measurements with the MINI-PAM-II: http://www.walz.com/products/chl_p700/mini-pam-II/downloads.html and the manual for the FireStingO2 oxymeter: (http://www.pyro-science.com/).
  • Page 100 Chapter 7 Touchscreen Operation Auto Range in the active state ignores the integration time set and determines the optimal integration time automatically. Operation Mode opens another menu (Fig. 59, page 94) in which the type of spectrum (light, fluorescence or reflectance) and, for light spectra, the entrance optics (flat versus spherical) and the environment in which measurements are performed (air or water) can be specified.
  • Page 101 Touchscreen Operation Chapter 7 the detector under water. The last two items in the Setup menu allows adjusting the intensity of the blue and green LED used as excitation source for fluorescence emission spectra. Limits: In the menu “Limits” short and long wavelength borders of reflectance spectra can be defined, and also the short wave- length limits of fluorescence emission spectra.

  • Page 102: Load System Settings

    “”Load Defaults” (Fig. 62, page 96). 7.3.5 MINI-PAM-II Settings The menu “MINI-PAM-II Settings” permits adjusting of fluorome- ter settings, choosing between two time intervals for the Ft chart settings, and retrieving the default configuration for measure- ments.
  • Page 103 Percentage of maximum intensity of the display’s backlight LED array. Signal LED On/off switch for LED on top of the MINI-PAM-II (green flash eve- ry 2 s, normal operation; green double flash every 2 s, clock- controlled operation; continuous light, saturation pulse analysis;...
  • Page 104 Chapter 7 Touchscreen Operation Fig. 63: MINI-PAM-II, Memory and Info menus...
  • Page 105 Touchscreen Operation Chapter 7 Table 9: Default Settings Default Setting Current Setting (saved and restored) Measuring Light Status Intensity 6, relative unit Frequency 3, see Table 8 Frequency high status Actinic Light Status Intensity 6, relative unit Factor 1.00 PAM Signal Gain 1, relative unit Damping...
  • Page 106 Chapter 7 Touchscreen Operation Table 9: Default Settings Default Setting Current Setting (saved and restored) Hardware Signal LED status Beeper status Automatic power down 15, min Background light Graphics Ft chart time resolu- 0.2, s/dot tion External Light Status Total intensity Red LED Green LED Blue LED...

  • Page 107: Memory

    63), respectively. Also in this window, new records can be started and the mark of saturation pulse data can be changed. Deletion of data from the MINI-PAM-II internal memory is not al- lowed in the stand-alone mode. However, the memory can be cleared using the software WinControl-3.

  • Page 108: Mini-Pam-Ii Info

    Chapter 7 Touchscreen Operation MINI-PAM-II provides hardware information of the fluorome- ter. Sensors lists serial numbers of sensors connected to the MINI-PAM-II. Firmware shows serial number and date of the firmware of the MINI-PAM-II. Info Side Panel MINI-PAM-II  ...

  • Page 109: Sensor Info

    23.6 Temp. Detector (°C) EXIT Operation Time (h ) Fig. 66: MINI-PAM-II Info 7.3.7.2 Sensor Info Serial number and hardware information of MINI-PAM-II measur- ing head and accessories. Side Panel Sensor Info MINI-PAM Head (red/12)   . Up/down Leaf Clip (BLKC0255/30) keys move cursor.

  • Page 110: Firmware Info

    Chapter 7 Touchscreen Operation 7.3.7.3 Firmware Info Version and times of completion of firmware. Side Panel Firmware Info Version 136/1573   . Up/down Build 18-03-28 13:59:44 keys move cursor. Prog 18-03-29 09:33:46 EXIT. Return to Info. EXIT Fig. 68: Firmware Info...

  • Page 111: Specifications Mini-Pam-Ii

    µmol m MINI-PAM-II/R (Red Version) Measuring light: Red (655 nm) LED, modulation frequencies and PAR as described for MINI-PAM-II/B. Fluorescence at wave- lengths greater than 700 nm is measured Actinic light: Same red LED as for measuring light, maximum PAR of actinic light and saturation pulses as described for MINI-...
  • Page 112 Chapter 8 Specifications Data memory: Flash memory, 8 MB, providing memory for more than 27,000 saturation pulse analyses Display: Backlit 160x104 dots (78 x 61 mm) transflective B/W LCD display with resistive touchscreen Ports: Ports for fiberoptics, USB cable, external light source, 2035-B leaf clip, auxiliaries and 12 V DC power supply Power supply: 6 AA (Mignon) rechargeable batteries (Eneloop 1.2 V/2 Ah), providing power for up to 1000 yield measurements;...

  • Page 113: Fiberoptics Mini-Pam/F

    Dimensions: Active diameter 5.5 mm, outer diameter 8 mm, length 100 cm Weight: 180 g 8.1.3 Power Supply MINI-PAM-II/N Input: 100 to 240 V AC, 50 to 60 Hz Output: 12 V DC, 5.5 A Operating temperature: -5 to +45 °C, (non-condensing) Dimensions: 13 cm x 5.5 cm x 3 cm (L x W x H)

  • Page 114: Battery Charger 000190101101

    Chapter 8 Specifications 8.1.4 Battery Charger 000190101101 Input: 100 to 240 V AC, 50 to 60 Hz Output: 12 V DC, 1.0 A Operating temperature: -5 to +45 °C, (non-condensing) Dimensions: 17.5 cm x 10.5 cm x 3 cm (L x W x H) Weight: 300 g including cable 8.1.5 Distance Clip 60°...

  • Page 115: Transport Case Mini-Pam/T

    Programmed Features: Automatic determination of signal offset for all light intensities and all gain levels. Automatic calibration of internal PAR sensor against an external PAR sensor connected to the MINI-PAM-II Communication Protocol: USB Computer Requirements: Processor, 0.8 GHz. RAM, 512 MB.

  • Page 116: Fiberoptics Adapter 90° 2030-B90

    Humidity sensor: Humidity and temperature sensing integrated circuit, 0 – 100% relative humidity Power supply: MINI-PAM-II leaf clip socket (5 V/10 mA) Cable length: 180 cm Dimensions: 17 cm x 5.7 cm (max.) x 8 cm (max.) (L x W x H) Weight: 250 g (excluding cable) 8.2.2...

  • Page 117: Arabidopsis Leaf Clip 2060-B

    Specifications Chapter 8 quality is adjusted individually and the maximum PAR per light quality is 1500 µmol m Design: LED holder for 4 RGBW LEDs made of anodized alumi- num with cooling ventilator on the backside. The device can be fastened by two 7 cm long aluminum rods whose ends fit in drilled holes of the 2035-B clip Dimensions: 9 cm (max.) x 4.5 cm x 12 cm (max.) (L x W x H)

  • Page 118: Fiberoptics Holder For Surfaces 2060-A

    Chapter 8 Specifications 8.2.6 Fiberoptics Holder for Surfaces 2060-A Aluminum plate (6.0 x 3.3 cm max.) with 11 mm diameter circular hole (measuring area) and aluminum port to position fiber at an angle of 60° relative to the aluminum plate. With port for temper- ature sensor of 2065-M unit to measure surface temperature and thread to mount the PAR sensor of the 2065-M unit.

  • Page 119: Miniature Spectrometer Mini-Spec/Mp

    Specifications Chapter 8 8.2.8 Miniature Spectrometer MINI-SPEC/MP Design: POM tube, at one side, port for light detection, port for fluorescence excitation by blue (452 nm max) or green (525 nm max) LEDs, and port for white light from a tungsten lamp for re- flection measurements;...

  • Page 120: Par Calibration Block 000160101439

    Cuvette: Round stainless steel cuvette (7.5 mm wide, 9.0 mm deep) with top window adapter for connecting the fiberoptics of the MINI-PAM-II; embedded in POM body with injection port for microliter syringes and hose nozzles for connecting an external flow-through water bath (not included). Including three 6.0 x 1.5 mm magnetic stir bars Dimensions: 11.5 cm x 9 cm x 8.5 cm (L x W x H, including...

  • Page 121: Magnetic Stirrer With Fiberoptics Holder Mks-2500

    Specifications Chapter 8 8.2.10 Magnetic Stirrer with Fiberoptics Holder MKS-2500 Magnetic stirrer: To drive the magnetic stir bar in the Suspen- sion Cuvette KS-2500; with Plexiglas ring for centering the cu- vette and miniature stand to fix the fiberoptics on top of the cu- vette 8.2.11 Compact Tripod ST-2101A...

  • Page 123: Warranty

    (date on invoice). Conditions This warranty applies if the defects are called to the attention of Heinz Walz GmbH, Germany, in writing within two (2) years of the shipping date of the product. This warranty shall not apply to...

  • Page 124: Instructions

    The Warranty Registration form must be completed and returned to 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.

  • Page 125: Index

    Index Chapter 10 Index Back Light ........97 2030-B90 ......... 14 Barcode Scanner ......28 2035-B Leaf-Clip Basic Data ......... 52, 53 External Light Source ....20 Battery ......... 4, 11, 106 Signal LED ........ 16 Battery Voltage ......102 2054-L External LED Source ...
  • Page 126 Fig. 24 SAT Kinetics ....47 Far Red Settings ......78 Fig. 25 Touch Screen Calibration Figures ..........49 Fig. 01 MINI-PAM-II Rear View ... 8 Fig. 26 Principal Screen Layout .51 Fig. 02 MINI-PAM-II, Spectra ..10 Fig. 27 Basic Data, Change Marker .........53 Fig.
  • Page 127 Fo’ Mode ......... 75 Fig. 54 Used Cal. Profile ... 90 F-Offset ........... 75 Fig. 55 Leaf Clip Channel 2 Freezing of MINI-PAM-II ....48 settings ........ 91 Ft Chart ........... 57 Fig. 56 External Tem. Sesnor ..92 Ft Chart Resolution ......96 Fig.
  • Page 128 Micro-Quantum-Sensor ..... 14 Miniature Spectrometer ..22, 24, 26, 93 PAR Calibration ......91 MINI-PAM/F1 Miniature Fiberoptics .29 Thermocouple ......15 MINI-PAM-II Info ....102, 103 LED Spectra ........10 MINI-PAM-II Settings .......96 LED Temperature ......102 MINI-SPEC/MP ....22, 24, 26 Light Curve ........ 64, 83 Light Curves ........
  • Page 129 Index Chapter 10 On button ........... 9 Recovery Curve ......64 Operating Temperature ....106 References Operation Time ......102 Fluorescence Ratio Parameters 35 Optode ........92, 93 Fluorescence Reviews ....43 Optoelectronic Unit Light Curves ......42 Back Panel ........8 Saturation Pulse Analysis ..
  • Page 130 Chapter 10 Index Spectrometer ........58 Temperature ......15, 55 Spectrometer Settings ..... 93 Temperature Calibration ....91 Surface Clip ........18 Thermocouple ........15 Suspension Cuvette ......27 Time ..........97 SYNC ..........9 Top Level Windows ....50, 51 System Hangs ......... 48 Touchscreen Calibration ....49 System Overview ......

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