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Walz MINI-PAM Manual

Portable chlorophyll fluorometer
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PHOTOSYNTHESIS YIELD
ANALYZER MINI-PAM
Portable Chlorophyll Fluorometer
Handbook of Operation
2.115 / 04.96
2. Edition: August 1999
minip_1ea.doc
 Heinz Walz GmbH, 1999
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
Printed in Germany

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  • Page 1 Handbook of Operation 2.115 / 04.96 2. Edition: August 1999 minip_1ea.doc  Heinz Walz GmbH, 1999 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 Printed in Germany...

  • Page 3: Table Of Contents

    1.1 General safety instructions ............1 1.2 Special safety instructions............1 2 General Information ..............2 3 Basic Operation of the MINI-PAM..........4 4 Description of the eight Keyboard Functions ......7 4.1 Single key operations ..............7 4.2 Double key operations............... 8 5 Important Points for Correct YIELD-Measurements ....
  • Page 4 CONTENTS 9 Data Transfer................48 10 Operation of the MINI-PAM via a PC-Terminal and the RS 232 Interface ................. 51 11 Maintenance................53 11.1 Internal battery and its replacement ........53 11.2 Halogen lamp and its replacement .......... 54 11.3 Fuse replacement..............55 11.4 EPROM and its replacement ...........
  • Page 5 13.1 Technical specifications ............87 13.2 List of warnings and errors............91 13.3 PIN-assignments..............93 13.4 List of commands for operation of MINI-PAM via PC-terminal by user-written software ........94 13.5 Selected reviews on chlorophyll fluorescence and related topics ..................98 14 Rechargeable battery ...............

  • Page 7: Safety Instructions

    2. The MINI-PAM employs high intensity light sources which may cause damage to the eye. Avoid looking directly into these light sources during continuous illumination or saturation pulses.

  • Page 8: General Information

    (ETR) is calculated. In addition to this central information, the MINI-PAM also provides the possibility of measuring fluorescence quenching coefficients (qP, qN, NPQ), applying continuous actinic light for measurement of induction...
  • Page 9 For this purpose, the Chart-window is particularly useful, as it records all fluorescence changes like a chart recorder. This manual deals mainly with the MINI-PAM as such, operated as a stand alone unit. A separate manual will be provided for the...

  • Page 10: Basic Operation Of The Mini-Pam

    BASIC OPERATION OF THE MINI-PAM 3 Basic Operation of the MINI-PAM The MINI-PAM is very easy to operate. It has a two-line LC- display and a small tactile keyboard with eight function keys (ON, OFF, MODE, MEM, ∧, ∨, START, SET). In order to get started, only the fiberoptics have to be connected and the ON-key is pressed.
  • Page 11 RS 232 interface or transferred on a PC for further analysis. The MINI-PAM has been pre-programmed at the factory with standard settings (see list in 7.1) for all relevant measuring parameters (for example Measuring Light Intensity, Gain, Damping, Saturation Pulse Intensity, Saturation Pulse Width etc.).
  • Page 12 CHAPTER 3 BASIC OPERATION OF THE MINI-PAM combination with the SET-key. Details are given in the MODE-menu list (see 7.2).

  • Page 13: Description Of The Eight Keyboard Functions

    4 Description of the eight Keyboard Functions Fig. 1: Photosynthesis Yield Analyzer MINI-PAM Single key operations To switch MINI-PAM on (short pressing of the key). To activate the backlighting of the display (switches automatically off when no key operation for 50 s; power saving for field use);...

  • Page 14: Double Key Operations

    CHAPTER 4 DESCRIPTION OF KEYBOARD FUNCTIONS To enter the MEMORY-level of stored data with the last stored data set being displayed. ∧, ∨ To select one of 51 points of the MODE-menu or one of 4000 data sets when MEMORY is activated: To change a particular parameter setting in the MODE- menu after operating the SET-key.
  • Page 15 To reset program, if MINI-PAM for some reason SET+OFF does not respond to key-operations. If the MINI-PAM is switched on by RS 232-access the key- controller may not respond. In this case push the ON-key once. Note:Whenever a command is given which involves the switching on and off of the actinic halogen light source, a short beep- sound confirms that the command is carried out.

  • Page 16: Important Points For Correct Yield-Measurements

    IMPORTANT POINTS 5 Important Points for Correct YIELD- Measurements The main purpose of the MINI-PAM is the reliable determination of the YIELD-parameter ∆F/Fm (Genty-parameter). This task is carried out by the MINI-PAM with exceptional sensitivity and reproducibility. Because of the central importance of this particular type of measurement, a special section is devoted to it in this handbook (see section 12.3).
  • Page 17 CHAPTER 5 IMPORTANT POINTS (...E). The plot of ETR vs. PAR corresponds to a light-response curve of photosynthesis (see 12.3.9). 4) When YIELD is measured under field conditions, it is essential that the leaf position and effective PAR are not inadvertently changed.

  • Page 18: Description Of The Memory-Function

    CHAPTER 6 DESCRIPTION OF THE MEMORY-FUNCTION 6 Description of the Memory-Function All data recorded via START are automatically stored in RAM- memory with a capacity of 4000 data sets. They can be recalled on display via the MEM-key. Then, for example, the following information is shown: MEM 382: 12:27 27/MAY/95 322Y 21.1E...
  • Page 19 CHAPTER 6 DESCRIPTION OF THE MEMORY-FUNCTION (MODE-menu point 25) had been carried out beforehand (see 12.3.4). Further operation of SET (2x) leads back to the original display with time and date. Using the arrow keys ∧ and ∨ one can move within the memory and display any previously recorded data sets.

  • Page 20: The Mode-Menu

    CHAPTER 7 THE MODE-MENU 7 The Mode-Menu The MODE-menu contains 51 items corresponding to a variety of measured values, instrumental settings or special commands. The positions of the various menu points were arranged for optimal practicability, with the most frequently used functions being closest to the standard position 1.

  • Page 21: List Of Menu Points

    CHAPTER 7 THE MODE-MENU List of Menu points The Menu points are organized in functional blocks. The starting point of each block can be reached successively by simultaneous pressing of MODE and SET. The frequently used positions MARK, MEAS-INT and GAIN can be readily selected by going backwards from position 1 using the ∨-key.
  • Page 22 CHAPTER 7 THE MODE-MENU 18. L.CURVE+REC:OFF(SET) 19. LC-WIDTH 0:10 (SET) 20. LC-INT: 3 (SET) 21. IND.CURVE: OFF(SET) MODE+∨ 22. IND.C+REC: OFF(SET) 23. IND-DELAY 0:40 (SET) 24. IND-WIDTH 0:20 (SET) 25. Fo and Fm (SET) 26. qP and qN (SET) 27. NPQ (SET) 28.

  • Page 23: Description Of The Mode-Menu Points

    Description of the Mode-menu points The following list briefly describes the items contained in the MODE-menu, some of which are outlined in more detail in section 12.3 (Assessment of photosynthesis yield with the MINI-PAM). Standard settings are shown. Standard menu-position for display of the 445F 1739M 19.9C...
  • Page 24 CHAPTER 7 THE MODE-MENU Switch between the standard measuring 4: M.FREQ: (SET) F: 448 745Y 6.2E pulse frequency of 0.6 kHz (LOW) and 20 kHz (HIGH). At 20 kHz the signal/noise is increased by a factor of 5-6. On the other hand, at this high frequency the measuring light intensity can induce substantial fluorescence changes.
  • Page 25 745Y 6.2E saving automatics which turn off the MINI-PAM after 4 min without key operation. It is advisable to disable the AUTO-OFF when the MINI-PAM is connected to an external power supply (via CHARGE-socket). Whenever the instrument is switched off manually, the AUTO-OFF function is enabled again (automatic reset to ON-position).
  • Page 26 CHAPTER 7 THE MODE-MENU meant to be turned on for extended periods of time, as this may lead to excessive internal heating. Therefore, the illumination periods are restricted (see menu point 14: ACT-WIDTH). There is a blinking sign (ACT) in the upper left corner while actinic illumination is on. On/off switch of the internal actinic light 13: ACT+YIELD: OFF(SET) F: 448...
  • Page 27 CHAPTER 7 THE MODE-MENU can be also directly started by double key operation ON + ∧. The time periods at the different intensities are set by menu point 19: LC- WIDTH. There is a blinking sign (LC) in the upper left corner of the display while a LIGHT CURVE is recorded.
  • Page 28 CHAPTER 7 THE MODE-MENU sufficient for so-called "rapid light curves". It is limited to 3 min in order to avoid excessive internal heating. The LC-INT determines the starting LC-INT: 3 (SET) F: 448 745Y 6.2E intensity which can be chosen between settings 1 to 5.
  • Page 29 CHAPTER 7 THE MODE-MENU fluorescence, Fo, and maximal fluorescence, Fm, of a dark-adapted sample by use of the SET-key. The thus sampled values are stored until new values are sampled via SET. With START a normal YIELD-determination is carried out and the given Fo- and Fm-values are maintained.
  • Page 30 CHAPTER 7 THE MODE-MENU excess light energy which is actively dissipated by plants into heat in order to avoid photodamage. Contrary to qN, NPQ-determination does not require knowledge of Fo and is not affected by non- photochemical quenching of Fo. NPQ is set to 0.000 if M>Fm. On/off switch of repetition clock which 28: REP-CLOCK: OFF(SET) F: 448...
  • Page 31 (see in section 7.1). Number and date of origin of current 37: PROGR.M2.24(170299) F: 448 745Y 6.2E program version of the MINI-PAM which is resident on EPROM. Function to move the present MEMORY- MEMORY: 125 (SET) F: 448 745Y 6.2E...
  • Page 32 CHAPTER 7 THE MODE-MENU Note: The MEMORY-front is identical to the MEM-number under which the last data set was stored. It advances by 1 with each following YIELD-determination. Command to erase all data accumulated in 39: CLEAR MEMORY (SET) F: 448 745Y 6.2E MEMORY.
  • Page 33 CHAPTER 7 THE MODE-MENU Function adjustment leaf 43:TEMP.GAIN:1.00 (SET) F: 448 745Y 6.2E temperature-reading with optional Leaf- Clip Holder 2030-B. The adjustment via TEMP-GAIN should be carried out after a preceding adjustment by TEMP-OFFS (menu point 42) at a different temperature, such that the slope in the temperature response curve can be evaluated.
  • Page 34 CHAPTER 7 THE MODE-MENU time constants (defined for 63.2 % of a signal change): 1: 0.05 s, 2: 0.2 s, 3: 1 s. Setting electronic signal gain 49: OUT-GAIN: (SET) F: 448 745Y 6.2E (amplification factor) which can be varied between 1 and 12.

  • Page 35: Components Of The Mini-Pam

    The basic functional system for measurements of fluorescence yield and of the effective yield of photosynthetic energy conversion consists of the MINI-PAM Main Control Unit and the fiberoptics. Additional peripheral components can be connected to the four sockets at the rear side of the Main Control Unit. Fig. 2 shows a functional block diagram of the MINI-PAM and its most essential accessories.

  • Page 36: Fluorescence Excitation And Detection

    8.1.1 Fluorescence excitation and detection In the standard version of the MINI-PAM fluorescence is excited by pulse modulated red light from a light-emitting-diode (LED). The pulse-width is 3 µs and pulse frequency is 0.6 or 20 kHz. In the so- called "burst-mode"...

  • Page 37: Special Information On Mini-Pam/B

    Special information on MINI-PAM/B Recently strong blue LEDs with an emission peak around 470 nm have become available and the MINI-PAM/B was developed which employs such LED as measuring light source. Using 470 nm measuring light has a number of technical and practical consequences which shall be briefly outlined.
  • Page 38 (ETR) as well. As the red part of the spectrum is not used in the MINI-PAM/B, this aspect is less problematic than in the standard instrument version.

  • Page 39: Internal Halogen Lamp As Actinic Light Source

    CHAPTER 8 COMPONENTS OF THE MINI-PAM • Chlorophyll emission: As the cut-off wavelength of the detector filter is shifted from 710 nm in the standard version to 650 nm in the blue version, the latter also detects the main chlorophyll emission peaking around 685 nm which originates mainly from photosystem II and, hence, shows higher values of variable fluorescence.

  • Page 40: Rechargeable Battery

    MINI-PAM housing. For recharging, the Battery Charger MINI-PAM/L is provided which is connected to the CHARGE-socket at the rear side of the MINI-PAM. The charger, which operates at input voltages between 100 and 240 V AC, features an overload protection.

  • Page 41: Lc-Display

    CHAPTER 8 COMPONENTS OF THE MINI-PAM The MINI-PAM can be also powered by an external 12 V battery for which purpose a special cable (MINI-PAM/AK) is available which can be connected to the CHARGE-socket at the rear side of the MINI-PAM. It should be noted, that a recharging of the internal battery with a 12 V external battery is not possible.

  • Page 42: Electronic Components

    CMOS microcontroller. The program software is stored in a CMOS EPROM. This EPROM is readily accessible after removing the bottom of the MINI-PAM (see 11.4) and can be exchanged by the user, if program up-dates become available. A CMOS RAM with 128 kB serves as data memory, providing storage capacity for 4000 data sets.

  • Page 43: Description Of The Connectors

    LEAF CLIP RS 232 OUTPUT CHARGE Fig. 4 At the rear side of the MINI-PAM besides the optical fiber connector the following electrical connectors are located: a) LEAF-CLIP The LEAF CLIP socket can be used for connecting the Leaf-Clip Holder...

  • Page 44: Fiberoptics Mini-Pam/F And Miniature Fiberoptics Mini-Pam/F1

    MINI-PAM/F1 The fiberoptics are inserted into the corresponding adapter at the rear side of the MINI-PAM. The active cross section of the standard version MINI-PAM/F is 5.5 mm. A special version (MINI-PAM/F1) with Ø 2 mm is also available, consisting of a single plastic fiber. In the standard version, numerous 70 µm fibers are thoroughly...
  • Page 45 CHAPTER 8 COMPONENTS OF THE MINI-PAM Fig. 5 Two spacer rings may be used to define fixed distances. The fiberoptics exit plane is positioned at a 60° angle relative to the sample plane. In this way shading of the sample is minimized, if the fiberoptics are pointing towards the sample from the side opposite to incident light.
  • Page 46 The fibers are protected by a steel-spiral and plastic mantle which provides a natural resistance to strong bending. In addition to the standard fiberoptics MINI-PAM/F, with an active diameter of 5.5 mm, a miniature fiberoptics with 2 mm active diameter (MINI-PAM/F1) is available for small spot measurements.

  • Page 47: Leaf-Clip Holder 2030-B

    CHAPTER 8 COMPONENTS OF THE MINI-PAM light may show an actinic effect. This can be counteracted by use of the ML-BURST function (MODE-menu position 5, see 12.3.2). Because of the strong influence of sample distance on signal amplitude, particularly with the 2 mm Ø fiberoptics, it is recommended to clamp the fiberoptics tip at fixed distance to the sample surface.
  • Page 48 CHAPTER 8 COMPONENTS OF THE MINI-PAM Micro-quantum-sensor monitoring PAR This tiny sensor is unique in monitoring the photosynthetic active radiation (PAR) at the very spot where also fluorescence is measured and at which photosynthetic performance is assessed. This function already is fulfilled, when approximately 10 % of the total measuring area is occupied by the sensor.
  • Page 49 CHAPTER 8 COMPONENTS OF THE MINI-PAM diffuser clean. Also, it must be pointed out that there is some decrease in sensitivity when the sensor is moved from the center of the measuring field to its periphery. It is advisable to check calibration regularly by comparison with a standard quantum sensor, like the LI-190.
  • Page 50 Pressing the 'remote' control push button on the handle of the Leaf-Clip Holder 2030-B is equivalent to operation of START on the MINI-PAM keyboard. In practice, this offers the advantage, that the experimenter can use both hands for positioning the leaf within the holder and at the same time trigger a recording by remote control.

  • Page 51: Micro Quantum/Temp.-Sensor 2060-M

    MINI-PAM housing. A 20 W lamp is powered by an external battery (e. g. NP-3/12). Its intensity can be varied steplessly via a 15-turn potentiometer.
  • Page 52 With the 15-turn potentiometer defined PAR-values can be chosen, which are read off the MINI-PAM LC-display. A switch is provided to turn the lamp on/off. A major application of the External Halogen Lamp 2050-HB is the adjustment of defined light intensities for measurements of light saturation curves under field conditions.

  • Page 53: Dark Leaf Clip Dlc-8

    CHAPTER 8 COMPONENTS OF THE MINI-PAM Dark Leaf Clip DLC-8 The Dark Leaf Clip DLC-8 weighs approx. 4 g and, hence, can be attached to most types of leaves without any detrimental effects. It is equipped with a miniature sliding shutter which prevents light...

  • Page 54: Data Transfer

    9 Data Transfer Since the introduction of the WinControl software, the normal way of data transfer from the MINI-PAM to a PC is via a special routine provided by WinControl (see separate manual). In addition, two other programs are provided for the transfer of data from the MINI-PAM via RS-232 interface cable to a PC.
  • Page 55 'Fist Data', after MEM 4000 the transfer continues from MEM1 upwards. Update versions of the MINI-PAM Data Transfer Program can be downloaded from our website http://www.walz.com. In this case, relevant information concerning the update will be contained in a Read-me file.
  • Page 56 CHAPTER 9 DATA TRANSFER Fig. 7: WINDOWS version of software PAMTRANS...

  • Page 57: Operation Of The Mini-Pam Via A Pc-Terminal And The Rs 232 Interface

    WinControl software has distinct advantages, particularly with respect to data display (see separate WinControl manual). In addition, there is the possibility to connect the MINI-PAM via the RS 232 interface to a PC and to control all functions by PC-keyboard operations using user-written software.
  • Page 58 Fm', as measured with the last saturation pulse, can be called on display. In this way, it is possible to carry out all MINI-PAM functions by remote control from a PC terminal and to transfer information from the MINI-PAM to a PC. In principle, using the TERMINAL-program...

  • Page 59: Maintenance

    In this case, battery replacement is recommended. The MINI-PAM features a number of functions and warnings which make it highly unlikely that excessive discharge of the battery occurs inadvertently: •...

  • Page 60: Halogen Lamp And Its Replacement

    External Halogen Lamp 2050-HB in combination with the Leaf Clip Holder 2030-B. For replacement of the internal halogen lamp the MINI-PAM is opened by removing the bottom part (4 screws). The lamp is held in a pre-focused position by an aluminum mounting-frame, which is fastened to the optical compartment by two screws.

  • Page 61: Fuse Replacement

    EPROM which is directed to the side of the MINI-PAM housing where the various connectors are. For lifting the EPROM, a paper-clip can be useful. Put a finger on it, so that it does not jump up.
  • Page 62 CHAPTER 11 MAINTENANCE Fig. 8...

  • Page 63: Chlorophyll Fluorescence Measurements With The Mini-Pam

    The MINI-PAM differs from the previously issued PAM fluorometers in that it is further miniaturized and optimized in order to perform one particular type of measurement with the greatest ease, accuracy and reliability, namely the determination of the effective quantum yield of photosynthetic energy conversion, ∆F/Fm', the so-...

  • Page 64: Chlorophyl Fluorescence As An Indicator Of Photosynthesis

    Genty-parameter. In the following sections some background information on this and other fluorescence parameters is given, and special aspects on fluorescence measurements with the MINI-PAM are outlined, in order to make optimal use of this instrument. 12.1 Chlorophyl fluorescence as an indicator of photosynthesis...
  • Page 65 CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM Fig. 9: Schematic view of primary energy conversion and primary electron transport in photosynthesis. LHC, light harvesting pigment-protein complex; P680 P700, energy converting special chlorophyll molecules in the reaction centers of photosystem II (PSII) and photosystem I (PSI), respectively;...
  • Page 66 CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM 'quenching analysis'. For this purpose the PAM measuring principle has been developed which allows monitoring fluorescence against times larger background signals (see 12.2). From the viewpoint of fluorescence emission there are two fundamentally different types of competing de-excitation processes: •...
  • Page 67 This is an important point for field investigations, for which the MINI-PAM was optimized. In recent years, evidence from a number of research groups has...
  • Page 68 B. The difference could as well arise from leaf A being exposed to stronger light or to a lower temperature than leaf B. The MINI-PAM offers the possibility to measure photosynthetically active radiation (PAR) and temperature...
  • Page 69 CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM 8.3), such that together with every YIELD-value also the corresponding values of PAR and temperature are entered into the file of automatically stored data. When PAR is known, the apparent rate of electron transport (ETR) is calculated (displayed as ...E).

  • Page 70: The Pam Measuring Principle

    (see also Fig. 11): Fluorescence is excited by very brief but strong light pulses from light-emitting diodes. With the MINI-PAM, these pulses are 3 µs long and repeated at a frequency of 600 or 20000 Hz. The LED light passes a short-pass filter (λ<670 nm) and the photodetector is...
  • Page 71 Due to the PAM measuring principle saturation pulse induced fluorescence changes can be very selectively and reliably analyzed in terms of photosynthetic activity. With the MINI-PAM, just like with all PAM fluorometers, even small values of ∆F induced by a saturation pulse can be relied on.
  • Page 72 CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM displayed. Such reliable performance, which is not possible with conventional amplifier systems, is of particular importance when photosynthesis yield is low due to stress conditions. In such cases it is essential to be sure that total inhibition really is indicated by YIELD = 0.000.

  • Page 73: Assessment Of Photosynthesis With The Mini-Pam: Outline Of The Most Important Functions In Practical Applications

    12.3 Assessment of photosynthesis with the MINI-PAM: Outline of the most important functions in practical applications. As soon as the MINI-PAM is switched on, it continuously monitors the fluorescence yield of a sample which is close to the fiberoptics exit. In section 12.1 it was outlined, in which way fluorescence yield relates to the effective quantum yield of photochemical energy conversion.

  • Page 74: Ml-Burst (Menu Point 5)

    . Even if a sample is kept in absolute darkness, the actual fluorescence measurement requires some excitation light. With the MINI-PAM, under standard conditions this amounts to ca 0.15 µmol quanta m . It can be decreased by lowering the measuring light intensity (menu point 50: MEAS-INT) or by applying the 'burst mode' (menu point 5: ML-BURST).

  • Page 75: Auto-Zero (Menu Point 2)

    CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM 12.3.3 AUTO-ZERO (menu point 2) The MINI-PAM, like any other chlorophyll fluorometer, is not absolutely selective for chlorophyll fluorescence but also shows a small signal when no plant sample is in contact with the fiberoptics.

  • Page 76: Fo, Fm (Menu Point 25)

    CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM applying maximal measuring light intensity and maximal gain. However, in this way also the background signal is increased and AUTO-ZERO becomes very essential. For example, in an experiment with a 1 mm² piece of a leaf at maximal gain and measuring light intensity quite reproducibly an Fv/Fm = 0.610 to...

  • Page 77: Int.temp (Menu Point 35)

    Hence, any internal heating of the fluorometer will lead to a corresponding decrease of the signal amplitude (see section 12.3.5). To take this aspect into account, the MINI-PAM features measurement of internal temperature (menu point 35), which is automatically registered with every YIELD-measurement and stored in MEMORY.
  • Page 78 0.6 kHz. Changes in measuring light intensity induced by temperature changes, just like any effect on the sensitivity and selectivity of fluorescence measurements with the MINI-PAM, can be also evaluated monitoring fluorescence...

  • Page 79: Qp, Qn And Npq (Menu Points 26 And 27)

    CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM illumination and longer saturation pulses can induce temperature increases within the fluorescence standard of up to 10 °C, corresponding to a decrease of fluorescence yield of 4 %. Actually, the heating during strong illumination also has a small effect on leaf measurements, as also chlorophyll fluorescence is lowered by approximately 0.4 % per °C.
  • Page 80 PS I, can enhance Q -reoxidation and facilitate assessment of Fo'. However, the MINI-PAM does not feature an intrinsic far-red light source (as e.g. the PAM-2000). Therefore, it should be realized that the measured values of qP and qN are valid in first approximation only, in particular when strong energy-dependent nonphotochemical quenching is given.

  • Page 81: Yield-Measurements Of Illuminated Samples

    12.3.7 YIELD-measurements of illuminated samples With every application of START a YIELD-measurement is carried out and on the LC-display of the MINI-PAM in the standard menu position 1 the following signals measured in connection with a particular saturation pulse are shown: •...

  • Page 82: Act-Light And Act+Yield (Menu Points 12 And 13)

    In one of its most common applications, the MINI-PAM assesses YIELD of plants in their natural light environment under steady-state conditions. In this case, use of the Leaf-Clip Holder 2030-B with automatic measurement of PAR and leaf temperature is very convenient (see 8.3).
  • Page 83 ON + SET or in menu position 12 via SET. In the latter case the remaining illumination time is displayed. The MINI-PAM also features the possibility of combining actinic illumination and YIELD-determination. In menu position 13: ACT + YIELD, there is first actinic illumination and at the end of the chosen period a saturation pulse is applied for YIELD-determination.
  • Page 84 LIGHT CURVE recordings (see 12.3.9), the ACT- WIDTH should be small, in order to avoid excessive internal heating of the MINI-PAM. In these applications it is limited to 3 min. The ACT + YIELD function provides very essential information on the state of the photosynthetic apparatus of a sample.

  • Page 85: Light Curve (Menu Point 17) And Light-Curve+Rec (Menu Point 18)

    CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM the overall yield of assimilation is equivalent to the yield of energy conversion at PS II. For a limitation to become apparent, the system must be 'put under light pressure'. For example, if some stress factor...
  • Page 86 CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM instrument overheating with longer illumination times. An alternative possibility to vary the ACTINIC INTENSITY range is given by the ACT-FACT function (menu point 16). This shifts all intensities up or down. Before starting a LIGHT CURVE recording, a sample should be well adapted to a moderate light intensity, which is close to the light intensity experienced by the plant in its natural environment.
  • Page 87 PS II (so-called State Shift). The apparently irreversible YIELD-lowering (with respect to the original dark state) is expression of "photoinhibition". LIGHT CURVES as measured with the MINI-PAM contain somewhat different information than the conventional light response curves. Correct measurement of the latter requires the attainment of steady state at each PAR-value, which takes at least 10 min.

  • Page 88: Yield- And Etr-Averaging (Menu Point 11)

    12.3.10 YIELD- and ETR-averaging (menu point 11) With normal samples under standard conditions the signal/noise ratio obtained with the MINI-PAM is rather high, such that a single measurement results in the reliable determination of YIELD and ETR (see 12.3.7). In practice, the averaging function is most useful in order to obtain representative information on the photosynthetic performance of a heterogeneous sample.

  • Page 89: Induction Curve (Menu Point 21) And Induction Curve+Recovery (Menu Point 22).... 83

    YIELD approaches zero, while ETR may still be substantial due to high PAR-values. Under such conditions, even the MINI-PAM becomes limited by the signal/noise ratio in YIELD-determination, which can be improved by averaging.
  • Page 90 Induction curves are either recorded via the analog output of the MINI-PAM using a chart recorder or via the RS 232 interface using a PC under WinControl-software. The latter offers the possibility of online registration and display of various derived fluorescence parameters, like effective quantum yield and quenching coefficients (see separate WinControl manual).

  • Page 91: Repetition Clock (Menu Point 28: Rep-Clock And Double Key Function On+Mem)

    CHAPTER 12 MEASUREMENTS WITH THE MINI-PAM on all features of the the induction curves. This aspect is particularly relevant for endosymbiotic phycobionts, as O is consumed by their own and the host's dark-respiration and O -diffusion is restricted (see recent report by Schreiber, Gademann, Ralph and Larkum:...
  • Page 92 In this context it is important to note that the full capacity of a freshly charged battery allows approximately 12 hours continuous operation of the MINI-PAM with standard YIELD- determination every minute (total of ca. 720 saturation pulses). When...

  • Page 93: Appendix

    CHAPTER 13 APPENDIX 13 Appendix 13.1 Technical specifications Photosynthesis Yield Analyzer MINI-PAM Measuring light source: Light emitting diode, emission maximum at 650 nm; 12 intensity settings, standard intensity 0.15 µmol m PAR; modulation frequency 0.6 or 20 kHz; Auto 20 kHz function; burst-mode, 1/5...
  • Page 94 CHAPTER 13 APPENDIX Display: 2 x 24 character alphanumerical LC- display with backlight; character size 4.5 mm User interface: 2 x 4 tactile keypad Power supply: Internal rechargeable battery 12 V/2 Ah, providing power for ca. 1000 yield measurements; automatic power/off when not used for 4 min;...
  • Page 95 Dimensions: Active diameter, 5.5 mm; outer diameter, 8 mm; length, 100 cm Weight: 0.18 kg Battery Charger MINI-PAM/L Power Supply: 100 to 240 V AC, 50/60 Hz Output: 18 V/45 W Dimensions: 13.5 cm x 6 cm x 3.6 cm (L x W x H) Weight: 0.26 kg...
  • Page 96 Ni-CrNi, ∅ 0.1 mm, -20 to 60 °C Thermocouple: Power supply: Via cable connecting to MINI-PAM Output: PAR and °C on LC-display of MINI-PAM; push-button trigger-signal to start saturation pulse for yield-measurement Weight: 0.35 kg External Halogen Lamp 2050-HB (optional) Wavelength: <710 nm...

  • Page 97: List Of Warnings And Errors

    Active diameter, 2 mm; length, 150 cm 13.2 List of warnings and errors Errors in MINI-PAM performance and warnings concerning sub- optimal use of the instrument are signalled by messages in the upper left corner of the display line. The following list briefly describes the various error messages: Err.
  • Page 98 Battery voltage has dropped below 11.2 V which means that only 20-30 further measurements are possible: Recharge battery or connect external battery by special cable (MINI-PAM/AK, optional). Err. ? NEW OFFSET ? Last measurement may be erroneous as GAIN (menu point...

  • Page 99: Pin-Assignments

    CHAPTER 13 APPENDIX Battery voltage has dropped below 11.2 V: Be prepared that the error message 3 (LOW BATTERY) will appear when START is applied. Actinic illumination is running. Actinic illumination with terminal YIELD-determination (menu point 12) is running. REPETITION-CLOCK (menu point 28) is running. Automatic recording of a LIGHT CURVE (menu point 17) is running.

  • Page 100: List Of Commands For Operation Of Mini-Pam Via Pc-Terminal By User-Written Software

    13.4 List of commands for operation of MINI-PAM via PC-terminal by user-written software As described in chapter 10, the MINI-PAM can be operated by remote control from a PC terminal. For this purpose a suitable TERMINAL-program must be installed and the RS 232 interface cable connected to the corresponding communication port.
  • Page 101 Hence, data transfer should not occur during measurements. In addition, also the WinControl software is available, which has been optimized for the communication between PC and MINI-PAM and features a number of most comfortable functions for data acquisition and analysis (see separate manual).
  • Page 102 CHAPTER 13 APPENDIX Command Corresponding Description point in MODE-menu dat or Date (day month year) dat(ddmmyy) dl1/dl0 Display light on/off Display MODE-menu point x ETR (electron transport rate) Averaged ETR (Leaf Clip) ec1/ec0 Echo on/off ef or efx ETR-factor (defined as x) Fluorescence yield before last sat.pulse, F Momentary fluorescence yield, F*...
  • Page 103 Display of data set Mem.x o+ or o+x Display of data sets from 1 to x Display of data set format MINI-PAM switched off Display of present MINI-PAM settings MINI-PAM settings reset to standard Display of present qN Display of present qp...

  • Page 104: Selected Reviews On Chlorophyll Fluorescence And Related Topics

    CHAPTER 13 APPENDIX Command Corresponding Description point in MODE-menu converter (x = 0 ... 7) YIELD measured with last sat. pulse Averaged YIELD No. of averaged YIELD-values Reset YIELD-averaging function 13.5 Selected reviews on chlorophyll fluorescence and related topics Allen JF (1992) Protein phosphorylation in regulation of photosynthesis.
  • Page 105 CHAPTER 13 APPENDIX Bolhar-Nordenkampf HR, Long SP, Baker NR, Öquist G, Schreiber U and Lechner EG (1989) Chlorophyll fluorescence as a probe of the photosynthetic competence of leaves in the field: a review of current instrumentation. Functional Ecology 3:497-514 Bose S (1982) Chlorophyll fluorescence in green plants and energy transfer pathways in photosynthesis.
  • Page 106 CHAPTER 13 APPENDIX Edwards GE and Baker NR (1993) Can CO assimilation in maize leaves be predicted accurately from chlorophyll fluorescence analysis? Photosynth Res 37:89-102 Falkowski PG and Kolber Z (1995) Variations in chlorophyll fluorescence yields in phytoplankton in the world oceans. Aust J Plant Physiol 22:341-355 Govindjee (1990) Photosystem II heterogeneity: the acceptor side.
  • Page 107 CHAPTER 13 APPENDIX Krall JP and Edwards GE (1992) Relationship between photosystem II activity and CO fixation in leaves. Physiol Plant 86:180-187 Krause GH and Weis E (1984) Chlorophyll fluorescence as a tool in plant physiology. II Interpretation of fluorescence signals. Photosynth Res 5:139-157 Krause GH and Weis E (1991) Chlorophyll fluorescence and photosynthesis: The basics.
  • Page 108 CHAPTER 13 APPENDIX Pfündel E and Bilger W (1994) Regulation and possible function of the violaxanthin cycle. Photosynth Res 42:89-109 Renger G (1992) Energy transfer and trapping in photosystem II. In: Barber J (ed) The Photosystems: Structure, Function and Molecular Biology, pp 45-99. Elsevier, Amsterdam Renger G and Schreiber U (1986) Practical applications of fluorometric methods to algae and higher plant research.
  • Page 109 CHAPTER 13 APPENDIX Schreiber U and Bilger W (1993) Progress in chlorophyll fluorescence research: major developments during the past years in retrospect. Progress in Botany 54:151-173 Schreiber U and Neubauer C (1990) O -dependent electron flow, membrane energization mechanism non- photochemical quenching chlorophyll...

  • Page 110: Rechargeable Battery

    CHAPTER 14 RECHARGEABLE BATTERY 14 Rechargeable battery The Photosynthesis Yield Analyzer MINI-PAM is equipped with a rechargeable sealed-lead acid battery. The life time is 1-3 years and it depends on the specific application. A 10 °C rise of the temperature will decrease battery life by approx.

  • Page 111: Warranty Conditions

    WARRANTY CONDITIONS 15 Warranty conditions All products supplied by the Heinz Walz GmbH, Germany, are warranted by Heinz Walz GmbH, Germany to be free from defects in material and workmanship for one (1) year from the shipping date (date on invoice).
  • Page 112 3. All products being returned for warranty service must be carefully packed and sent freight prepaid. 4. 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...

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