Instrument Description and Information for Users - …

1 Printed in Germany imaging -PAM M-series Chlorophyll Fluorometer Instrument Description and Information for Users / 5. revised Edition: March 2014 Heinz Walz GmbH, 2014 Heinz Walz GmbH Eichenring 6 91090 Effeltrich Germany Phone +49-(0)9133/7765-0 Telefax +49-(0)9133/5395 E-mail Internet CONTENTS I 1 Safety instructions .. 1 General safety instructions .. 1 Special safety instructions .. 2 2 Introduction .. 3 3 Components of the imaging -PAM MAXI-version .. 7 Control Unit IMAG-CG .. 9 LED-Array Illumination Unit IMAG-MAX/L and IMAG-MAX/LR .. 11 CCD Camera IMAG-K6 and objective K6-MAX .. 15 CCD Camera IMAG-K7 and objectives K7-MAX/Z and K7-MAX/S .. 17 Mounting Stand with Eye Protection IMAG-MAX/GS .. 19 Leaf Distance Holder IMAG-MAX/B .. 24 Notebook PC IMAG-PC.

2 25 Adapter IMAG-MAX/GWK .. 26 4 Components of the imaging -PAM MINI-version .. 27 Multi Control Unit IMAG-CG .. 27 MINI-Head LED-Array IMAG-MIN/B and IMAG-MIN/R .. 28 CCD Camera IMAG-K6 or IMAG-K7 .. 30 IMAG-MIN/GFP with IMAG-K6 .. 32 Leaf Holder IMAG-MIN/BK with Grip Holder .. 38 Adapter for GFS-3000 (IMAG-MIN/GFS) .. 42 ImagingWin software versions for various types of MINI-Version .. 44 5 Components of the imaging -PAM MICROSCOPY-version . 46 Multi Control Unit IMAG-CG .. 47 CCD Camera IMAG-K6 .. 47 Axio Epifluorescence Microscope .. 49 Reflector Modules .. 51 CONTENTS II Assembling of beam splitter and filters .. 52 Mounting of the reflector module .. 54 LED Modules .. 56 Adjustment of brightness by grey filters .. 56 Integration of LED modules into Axio.

3 58 Connecting LED modules with IMAG-CG .. 60 Switching LED modules for measurements .. 60 IMAG-RGB .. 60 6 How to get started .. 63 Connecting the cables .. 63 Software installation .. 64 Installation and Starting of ImagingWin .. 64 Installation of camera driver .. 66 First steps and examples of routine measurements .. 66 7 ImagingWin .. 81 8 IMAGINGWIN - System Operation .. 83 Definition of New Record .. 83 Fo, Fm .. 83 New Record .. 84 Measure .. 84 Functions applying to the View-mode .. 85 Light controls .. 87 9 IMAGINGWIN - Register Cards .. 90 Image-window .. 90 Different types of images .. 90 Current fluorescence yield, Ft .. 91 Dark fluorescence yield, Fo .. 91 Fluorescence yield, F .. 92 CONTENTS III Maximal fluorescence yield, Fm .. 92 Maximum fluorescence yield, Fm'.

4 93 Maximal PS II quantum yield, Fv/Fm .. 93 Effective PS II quantum yield, Y(II) .. 94 Quantum yield of regulated energy dissipation, Y(NPQ)95 Quantum yield of nonregulated energy dissipation, Y(NO) .. 96 Absorptivity, Abs.. 97 Apparent rate of photosynthesis, PS/50 .. 99 NIR light remission, NIR .. 100 Nonphotochemical quenching, NPQ/4 .. 101 Red light remission, R .. 102 Coefficient of nonphotochemical quenching, qN .. 103 Coefficient of photochemical quenching, qP .. 104 Coefficient of photochemical quenching, qL .. 105 Inhibition, Inh.. 106 Image capture and analysis .. 107 Measure Abs.. 107 Area of Interest, AOI .. 108 Select: Fluorescence or Live Video .. 110 Zoom .. 112 Cursor .. 113 Analysis .. 113 Kinetics window .. 116 Light Curve window .. 122 Report window .. 128 Settings window .. 132 Light parameters .. 133 CONTENTS IV Gain and Damping.

5 136 Absorptivity .. 137 Slow Induction parameters .. 138 Image Correction .. 138 Image Transformation .. 141 Battery .. 141 Display parameters .. 142 Go Speed .. 144 PS Limit .. 144 Inh. Ref. AOI .. 145 Yield Filter .. 145 Fm Factor .. 146 F Factor .. 149 Reset Default Settings, Open or Save User Settings .. 152 High Sens. window .. 153 Special SP-Routine .. 154 Fo 156 Fv/Fm Contrast Enhancement by Background Suppression156 RGB-Fit window .. 157 RGB Gain .. 160 Fit Correction .. 160 10 IMAGINGWIN - Menu Bar .. 163 File .. 163 Transfer FoFm .. 163 Using Skript files - Load Script/Run Script .. 163 Exit .. 171 Options .. 172 Ruler .. 172 CONTENTS V Scale .. 172 Info Icons .. 173 Mean over AOI .. 173 Define AOI-array geometry .. 175 Create AOI array.

6 176 Al-List .. 178 LED currents / PAR values .. 178 Light Calibration .. 180 Recalc .. 181 Transect .. 182 11 List of key commands .. 185 12 Technical specifications .. 186 Components used in all Versions .. 186 Control Unit IMAG-CG .. 186 IMAG-K7 .. 187 IMAG-K6 .. 187 Windows Software ImagingWin .. 187 Battery Charger 2120-N .. 188 Components specifically relating to Maxi-version .. 189 LED-Array Illumination Unit IMAG-MAX/L .. 189 LED-Array Illumination Unit IMAG-MAX/LR .. 190 Optional filter plate IMAG-MAX/F (only for IMAG-MAX/L!) .. 191 External 300 W Power Supply .. 191 K7-MAX/Z .. 191 K7-MAX/S .. 192 K6-MAX .. 192 K6-MAX/M and K7-MAX/M .. 193 CONTENTS VI Mounting Stand with Eye Protection IMAG-MAX/GS .. 193 IMAG- 194 194 Transport Box IMAG-MAX/T.

7 194 IMAG-MAX/GWK1 .. 195 Components specifically relating to MINI-version .. 195 IMAG-MIN/B .. 195 IMAG-MIN/R .. 196 IMAG-MIN/GFP .. 196 K7-MIN .. 197 K6-MIN .. 197 K6-MIN/FS .. 197 K7-MIN/M and K6-MIN/M .. 198 IMAG-S .. 198 IMAG-MIN/ST .. 198 199 IMAG-MIN/BK .. 199 IMAG-MIN/GFS .. 199 Components specifically relating to MICROSCOPY-versions199 IMAG-AXIOSCOPE .. 199 IMAG-L470M .. 200 IMAG-L625M .. 200 IMAG-RGB .. 200 IMAG-AX-REF .. 201 13 Warranty .. 202 Conditions .. 202 Instructions to obtain Warranty Service, .. 203 CHAPTER 1 SAFETY INSTRUCTIONS 11 Safety instructions General safety instructions 1. Read the safety instructions and the operating instructions first. 2. Pay attention to all the safety warnings. 3. Keep the device away from water or high moisture areas. 4. Keep the device away from dust, sand and dirt. 5. Always ensure there is sufficient ventilation.

8 6. Do not put the device anywhere near sources of heat. 7. Connect the device only to the power source indicated in the operating instructions or on the device. 8. Clean the device only according to the manufacturer s recommendations. 9. If the device is not in use, remove the mains plug from the socket. 10. Ensure that no liquids or other foreign bodies can find their way inside the device. 11. The device should only be repaired by qualified personnel. CHAPTER 1 SAFETY INSTRUCTIONS 2 Special safety instructions The imaging -PAM is a highly sensitive research Instrument which should be used only for research purposes, as specified in this manual. Please follow the instructions of this manual in order to avoid potential harm to the user and damage to the Instrument . Never use the Multi Control Unit IMAG-CG with more than one Measuring Head plugged in at the same time.

9 The imaging -PAM employs strong blue light for excitation of chlorophyll fluorescence, for driving photosynthetic electron transport and for transient saturation of photosynthetic energy conversion (Saturation Pulse method). In order to avoid harm to your eyes, please avoid looking directly into this light, particularly during Saturation Pulses. CHAPTER 2 INTRODUCTION 32 Introduction The imaging -PAM Chlorophyll Fluorometer is specialized for the study of two-dimensional heterogeneities of photosynthetic activity. The imaging -PAM M-series covers a wide range of applications. Large scale samples with areas exceeding multiwell plate format can be imaged as well as microscopically small samples at the level of single cells. MAXI-, MINI- and MICROSCOPY-versions have been issued that are operated with the same Multi Control Unit IMAG-CG.

10 Like all PAM fluorometers, the imaging -PAM applies pulse-amplitude-modulated measuring light for assessment of chlorophyll fluorescence yield. The same LEDs not only serves for generation of the pulse-modulated measuring light, but also for actinic illumination driving photosynthesis and for Saturation Pulses transiently saturating energy conversion at Photosystem II (PS II) reaction centers. The Saturation Pulse method provides a non-destructive means of analyzing the photosynthetic performance of plants. It allows to assess the quantum yield of energy conversion at PS II reaction centers, which is affected by numerous intrinsic and environmental parameters, like the physiological health, light conditions and various stress factors. Since the introduction of PAM fluorometry in 1985, a large amount of literature has been published on the practical use of this method in many fields of plant science.