PSI FluorPen FP 110, PAR-FluorPen FP 110, Monitoring Pen MP 100 Manual And User Manual

FluorPen FP 110

PAR-FluorPen FP 110

Monitoring Pen MP 100

Manual and User Guide

Please read this manual before operating this product

PSI, spol. s r. o., Drásov 470, 664 24 Drásov, Czech Republic FAX: +420 511 440 901, TEL: +420 511 440 011, www.psi.cz

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This document and its parts can be copied or provided to a third party only with the express permission of PSI.

The contents of this manual have been verified to correspond to the specifications of the device. However, deviations cannot be ruled out. Therefore, a complete correspondence between the manual and the real device canno t be guaranteed. The information in this manual is regularly checked, and corrections may be made in subsequent versions.

The visualizations shown in this manual are only illustrative.

This manual is an integral part of the purchase and delivery of equipment and its accessories and both Parties must abide by it.

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TABLE OF CONTENTS

1 Information Before Using FluorPen Device ..................................................................................................................... 6

2 Technical Specification .................................................................................................................................................... 7

3 General Information ....................................................................................................................................................... 9

3.1 Device description ............................................................................................................................................... 12

4 List of equipment .......................................................................................................................................................... 13

5 Care and Maintenance .................................................................................................................................................. 14

6 Principle of Measurement ............................................................................................................................................ 15

7 How to get started ........................................................................................................................................................ 17

7.1 Pulses description and setting ............................................................................................................................. 17

7.2 Measurement ...................................................................................................................................................... 19

7.3 Protocols explanation .......................................................................................................................................... 20

7.3.1 OJIP Protocol ................................................................................................................................................... 20

7.3.2 Non-Photochemical Quenching (NPQ) Protocol ............................................................................................. 21

7.3.3 Light Curve (LC) Protocol ................................................................................................................................. 24

7.4 Multiple Measurement ....................................................................................................................................... 27

8 Control menu tree ......................................................................................................................................................... 28

9 USB Connection ............................................................................................................................................................ 36

10 Bluetooth Connection .............................................................................................................................................. 37

10.1 Bluetooth Pairing ................................................................................................................................................. 37

11 FluorPen Software ................................................................................................................................................... 41

11.1 Software Installation ........................................................................................................................................... 41

11.2 Menu and Icon Explanation ................................................................................................................................. 42

11.2.1 Main Menu ................................................................................................................................................. 42

11.2.2 Menu Settings ............................................................................................................................................. 43

11.2.3 Menu Online Control .................................................................................................................................. 44

11.3 Data Transfer and Visualization ........................................................................................................................... 46

11.4 Firmware Update ................................................................................................................................................. 48

12 GPS Module ............................................................................................................................................................. 50

12.1 GPS / FluorPen Operation ................................................................................................................................... 50

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12.2 Data Download .................................................................................................................................................... 51

13 Warranty Terms and Conditions .............................................................................................................................. 52

14 Troubleshooting and Customer Support ................................................................................................................. 53

15 Appendix .................................................................................................................................................................. 54

15.1 Battery Pack for Monitoring Pen ......................................................................................................................... 54

15.1.1 Standard Battery Pack ................................................................................................................................ 54

15.1.2 Extended Temperature Range Battery Pack ............................................................................................... 55

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1 INFORMATION BEFORE U SING FLUORPEN DEVICE

Read this manual carefully before operating the device. If you are not sure about anything in the manual, contact the manufacturer for clarification.

By accepting the device, the customer agrees to follow the instructions in this guide.

Always follow corresponding manuals while working with the FluorPen device or doing the maintenance.

It is forbidden to interfere with the hardware or software of the FluorPen device in any way without previous agreement with the manufacturer.

The following table presents basic highlight symbols used in this manual.

Symbol

Description

Important information, read carefully.

Complementary and additional information.

Tab. 1 Used symbols.

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2 TECHNICAL SPECIFICAT ION

Measured / Calculated Parameters:

F0, Ft, Fm, Fm‘, QY, NPQ 1, NPQ 2, NPQ 3, OJIP, LC 1, LC 2, LC 3, PAR (measured as PPFD)*

Cosine correction of PAR sensor*:

Cosine corrected up to 80º angle of incidence

Linearity*:

Maximum deviation of 1 % up to per 10,000 µmol.m-2.s-1)

Saturating Pulse Illumination:

Adjustable from 10 to 100 % (up to 3,000 µmol.m-2.s-1)

Actinic Illumination:

Adjustable from 10 to 1,000 µmol.m-2.s-1

Measuring Illumination:

Adjustable from 10 to 100 % (up to 0.09 µmol.m-2.s-1 per pulse)

LED Emitter specification:

Blue (455 nm)

Detector Wavelength Range:

PIN photodiode with 667 to 750 nm bandpass filters

Communication:

USB and Bluetooth

FluorPen Software:

Windows 7 or higher compatible**

Memory Capacity:

Up to 16 Mb

Internal Data Logging:

Up to 149,000 data points

Display:

Graphical display

Keypad:

Sealed, 2-key tactile response

Keypad Escape Time:

Turns off after 8 minutes of no use

Power Supply:

Li-Ion rechargeable battery

Battery Life:

48 hours typical with full operation

Low Battery Detection:

Low battery indication displayed

Size:

134 mm x 65 mm x 33 mm

Weight:

188 g

Sample Holder:

Mechanical leaf clip (non-destructive for plants, effective for sample pre-darkening)

Operating Conditions:

Temperature: 0 to +55 ºC, 32 to 130 ºF; Relative humidity: 0 to 95 % (non-condensing)

Storage Conditions:

Temperature: -10 to +60 ºC, 14 to 140 ºF; Relative humidity: 0 to 95 % (non-condensing)

Warranty:

1 year parts and labor (see the last page of this Operation Manual for precise warranty conditions)

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* Only in PAR-FluorPen FP 110.

** Windows is a registered trademark of Microsoft Corporation.

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3 GENERAL INFORMATION

FluorPen FP 110 is a portable, battery-powered fluorometer that enables quick and precise measurement of chlorophyll

fluorescence parameters in the laboratory, greenhouse, or in the field owing to an inbuilt GPS module. It can be effectively used for studying photosynthetic activity, stress detection, herbicide testing, or mutant screening. Affordable price and straight-forward two-button operation makes the FluorPen a perfect tool for teaching photosynthesis. Because of its rapid measurement capability and large internal memory, the FluorPen is also an invaluable tool for large plantscreening programs.

PAR-FluorPen FP 110 includes all features of the FluorPen FP 110, i.e., it measures chlorophyll fluorescence parameters Ft, QY, NPQ, OJIP, and LC (QY). Besides that, the PAR-FluorPen incorporates an integrated Light Meter for direct digital readouts of Photosynthetically Active Radiation (PAR) in the range from 400 to 700 nm, the span in which plants use energy during photosynthesis. PAR is measured as Photosynthetic Photon Flux Density (PPFD), which is indicated by units of quanta (photons) per unit time per unit surface area. The sensor has a uniform response to photons withing the 400 700 nm waveband. Instant readouts are provided as average values of 20 measurements. Recalibration of the PAR sensor in the PAR-FluorPen device – recommended after 2 years of use.

Monitoring Pen MP 100 is a lightweight, portable and durable fluorometer. It is an extended version of the popular FluorPen. Due to its robust and weatherproof construction the Monitoring Pen is designed for long-term, unattended monitoring of chlorophyll ﬂuorescence parameters in lab or field experiments, even in rough environmental conditions. It is battery operated (internal or external battery) and it may optionally use a solar panel as a power supply.

Measured data are sequentially stored in the internal FluorPen, PAR-FluorPen or Monitoring Pen memory. New design of FluorPen series devices allows use both types of communication with a PC - USB and Bluetooth. Comprehensive FluorPen

1.1 software provides data transfer routines and many additional features for data presentation in tables and graphs.

FluorPen versions:

FluorPen FP 110/S

Equipped with a standard leaf-clip.

FluorPen FP 110/D

Adapted for use with detachable leaf-clips; leaf clips sold separately.

FluorPen FP 110/P (Fig. 1c)

Intended for autonomous use in indoor conditions (previously Monitoring Pen-S). It features a plastic case, measuring probe and thread for tripod attachment.

FluorPen FP 110/X

The “X“ version is mounted with custom-made leaf-clip.

PAR-FluorPen FP 110/S (Fig. 1a)

Measures in addition to standard fluorescence protocols also Photosynthetically Active Radiation (PAR) in the range from 400 to 700 nm. Equipped with a standard leaf-clip.

PAR-FluorPen FP 110/D (Fig. 1b)

Measures in addition to standard fluorescence protocols also Photosynthetically Active Radiation (PAR) in the range from 400 to 700 nm. Adapted for use with detachable leaf-clips; leaf clips sold separately.

Unless stated otherwise in this manual information regarding the FluorPen FP 110 are relevant also for the Monitoring Pen MP 100.

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PAR-FluorPen FP 110/X

Measures in addition to standard fluorescence protocols also Photosynthetically Active Radiation (PAR) in the range from 400 to 700 nm. The “X“ version is mounted with custom-made leaf-clip.

Fig. 1a) PAR-FluorPen FP 110/S. b) PAR-FluorPen FP 110/D. c) FluorPen FP 110/P.

Monitoring Pen versions:

Monitoring Pen MP 100-E (Fig. 2a)

Durable, battery-powered ﬂuorometer intended for autonomous use in field conditions. It features a waterproof metal case, measuring probe, thread for tripod attachment, external pack with batteries and the FluorPen

1.1 software for data collection and processing.

Monitoring Pen MP 100-A (Fig. 2b)

Submersible, battery-powered fluorometer intended for underwater measuring of fluorescence parameters (also autonomous). It features a waterproof case, measuring probe, and the FluorPen 1.1 software for data collection and processing. External battery pack with batteries is sold separately. Generally, version A is intended for maximum water depth up to 2 meters. The device is equipped with two buttons that allow direct control of the device (even under water). Version B can be used in deeper water (maximum 10 m). There are no buttons on the device. This version is intended for automatic measuring; the device control is enabled via commands preprogrammed in a PC (placed on shore). The device is usually fixed in static position under water.

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Fig. 2a) Monitoring Pen MP 100-E. b) Monitoring Pen MP 100-A.

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3. 1 DE V ICE DE S CR I PT I ON

Fig. 3 Device description.

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4 LIST OF EQUIPMENT

Carefully unpack the carton. You should have received the following items:

• FluorPen/Monitoring Pen

• Carrying Case

• Textile Strap for Comfortable Wearing

• FluorPen Operating Manual (on a USB flash disc)

• FluorPen software and driver (on a USB flash disc)

• USB cable

• Self-Adhesive Rubber Pads for Optics Protection (FP 110/S only)

• Detachable Leaf-clips (FP 110/D only)

Other Accessories or Optional Features (according to your specific order)

For USB connection you need to have the USB driver installed in your PC. You find the driver on the installation disk (USB driver folder). For more information go to

If any item is missing, please, contact PSI. Also check the carton for any visible external damage. If you find any damage, notify the carrier and PSI immediately. The carton and all packing materials should be retained for inspection by the carrier or insurer.

For customer support, please write to: support@psi.cz

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5 CARE AND MAINTENANCE

• Never submerge the device in water! (except Monitoring Pen MP 100-A)

• The device should not come in contact with any organic solvents, strong acids or bases.

• Keep the optical part clean and dry. If cleaning is needed, use soft, non-abrasive tissue.

• Battery life is approximately 48 hours when the FluorPen is operated continuously.

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6 PRINCIPLE OF MEASURE MENT

FluorPen is a fluorometer which can measure different fluorescence parameters of plants. It is equipped with a blue LED emitter (455 nm), optically filtered and precisely focused to deliver light intensities of up to 3,000 µmol.m-2.s-1 to

measured plant tissue (Fig. 4).

Fig. 4 Schema of FluorPen FP 110/S.

To use measurements of chlorophyll fluorescence to analyse photosynthesis, researchers must distinguish between photochemical quenching and non-photochemical quenching (heat dissipation). This is achieved by stopping photochemistry, which allows researchers to measure fluorescence in the presence of non-photochemical quenching alone. To reduce photochemical quenching to negligible levels, a high intensity, short flash of light is applied to the leaf. This transiently closes all PSII reaction centers, which prevents energy of PSII being passed to downstream electron carriers. Non-photochemical quenching will not be affected if the flash is short. During the flash, the fluorescence reaches the level reached in the absence of any photochemical quenching, known as maximum fluorescence Fm. The efficiency of photochemical quenching (which is a proxy of the efficiency of PSII) can be estimated by compa ring Fm to the steady yield of fluorescence in the light Ft and the yield of fluorescence in the absence of photosynthetic light F0. The efficiency of non-photochemical quenching is altered by various internal and external factors. Alterations in heat dissipation mean changes in Fm. Heat dissipation cannot be totally stopped, so the yield of chlorophyll fluorescence in the absence of non-photochemical quenching cannot be measured. See picture below (Fig. 5).

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Fig. 5 Chlorophyll fluorescence.

FluorPen measures:

Ft - Instantaneous Chlorophyll Fluorescence

Ft is equivalent to F0 if the sample is dark-adapted.

QY - Quantum Yield

QY is a measure of the Photosystem II efficiency. QY is equivalent to Fv/Fm in dark-adapted samples and to F

v '

/F

m '

in

light-adapted samples.

OJIP - Chlorophyll Fluorescence Induction Kinetics

The OJIP curves enable observing major changes that occur during exposure of a sample to high irradiance (see more in Chapter 7.3.1).

NPQ - Non-Photochemical Quenching

The NPQ protocol is the most typically used measuring approach to quantify photochemical and non-photochemical quenching. The measurement should be performed with a dark-adapted sample. More in chapter (see more in Chapter 7.3.2)

LC - Light Curve

Photosystem II Quantum Yield estimated from fluorescence that is measured sequentially in several different light levels (see more in Chapter 7.3.3).

PAR* - Photosynthetically Active Radiation

Photosyntetically Active Radiation measured as Photosythetic Photon Flux Density (PPFD).

* Only in PAR-FluorPen FP 110.

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7 HOW TO GET STARTED

For more detailed information on particular steps of FluorPen operation please refer to chapter 8.

The device can work on Li-Ion battery or is charged via USB cable from PC or grid. Monitoring Pen can be charged also from battery pack (see more in chapter 15.1)

The FluorPen is controlled using two buttons:

• Use the MENU key to scroll through sequential menu options on the digital display.

• Use the SET key to select a menu option based on cursor (>) position.

7. 1 PU L SES DE S CR I PT I ON AN D S ET T IN G

Flash pulse

This function serves for setting of measuring pulses intensity. The measuring pulses are weak pulses, which are able to induce the minimal chlorophyll fluorescence (F0 or Ft). It takes only 30 µs and the maximum intensity is 3,000 µmol.m-2.s-1. It means 30 µs * 3,000 µmol.m-2.s-1 = 0,09 µmol.m-2 per pulse is the maximal intensity of the flash pulse.

Super pulse

This function serves for setting intensity of the saturating pulse. Saturating pulse induces maximum chlorophyll fluorescence (Fm). 100 % of intensity equals approximately 3,000 µmol.m-2.s-1.

Actinic pulse

This function serves for setting intensity of measuring pulses. Actinic light is basically the ambient light in which the algae are growing. 100 % of intensity equals approximately 1,000 µmol.m-2.s-1.

Pulses used in predefined protocols:

Measurements based on fluorescence

Used pulses

F

t

Flash pulse

QY

Flash pulse, Super pulse

OJIP

Super pulse

NPQ protocols

Flash pulse, Super pulse, Actinic pulse

Light Curves

Flash pulse, Super pulse (Actinic pulse is preset)

Default setting of light intensities:

Flash pulse 30 % = Measuring flash pulse

Super pulse 80 % = Saturating pulse

Actinic pulse 300 µmol.m-2.s-1 (30 %) = Actinic light

Please note that those paramets are recommended by manufacturer and you can change them according to your needs.

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Setting of optimal intensities of pulses:

Flash pulse setting

The optimum value of Flash pulse you can identify with QY measurement. Before QY measurement is recommended to set the intensity of Super pulse on 80 %.

Please note that QY measurement should be performed with dark adapted sample. New sample is needed for every measurement (measured point of the leaf can be inhibited after measurement).

F0 is increasing linearly with growing intensity of Flash pulse. Increasing Flash pulse is not affecting background value.

The Flash pulse setting recommended by manufacturer is 30 %. You can increase the intensity of Flash pulse in case of

sample with very low chlorophyll density. Please note that high intensities of Flash pulse can cause undesirable “actinic effect”. In this case the Flash pulse initiates the photochemistry. The Flash pulse affects F0 and the QY value will be lower.

The Flash pulse intensity at which the highest value of QY is reached is optimal for your sample (Fig. 6).

Fig. 6 QY measurement performed with different intensities of Flash pulse. Optimal setting is highlighted in red rectangle.

Super pulse setting

Best way to recognized the optimal intensity of Super pulse is perform OJIP measurement.

Please note that OJIP measurement should be performed with dark adapted sample. New sample is needed for every measurement (measured point of the leaf can be inhibited after measurement).

The Super pulse setting recommended by manufacturer is 80 %.

Perform the OJIP measurement with different intensities of Super pulse. When the Fv/Fm value stops growing during the OJIP measurement with various Super pulse intensities, the Super pulse intensity is optimal for your samples (Fig. 7).

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Fig. 7 OJIP measurement performed with different intensities of Super pulse.

Actinic pulse setting

Intensity of Actinic pulse should correspond with cultivation light intensity or should be set according to application.

If you see Overflow on display during measurement please lower the intensity of used pulses.

In case of Low value on display during measurement please heighten the intensity of used pulses.

7. 2 ME A SUR E ME N T

No device calibration is needed before chlorophyll fluorescence measurement. Results of fluorescence measurement depend on device settings.

Fluorescence measurement with FluorPen is perform as follows:

• If you want to prepare dark adapted sample (prior for F0, QY, NPQ, LC) place the sample for defined period

on dark place or apply on the plant the detachable leaf-clip which is modified for dark-adaptation. Duration

of sufficient dark-adaptation depends on species, mostly varies between 10 and 15 minutes.

• Turn ON device – hold SET button for 1 sec.

• Place the dark-adapted leaf into the leaf-clip.

• Go to Measure > choose required parameter for example - QY.

• Press SET to confirm.

• FluorPen measures required parameter. If you choose protocol such as OJIP, LC or NPQ the display shows

the percentage of measurements.

• Value of required parameter appears on display after measuring. Result of OJIP, NPQ or LC protocol is a

graph, so all data you will see after data download using FluorPen Software.

• During the measurement all data are stored into the device memory.

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7. 3 PR O T OC OL S EX P LA N AT I ON

7.3.1 OJIP PROTOCOL

The FluorPen device offers the possibility to capture rapid fluorescence transient – OJIP, which occurs during exposure of photosynthesizing organisms to high irradiance. The FluorPen software enables data downloading to a personal computer and subsequent OJIP analysis.

The OJIP protocol includes the following extracted and technical parameters:

Abbreviation

Explanation

Bckg

Background

F0

F0 = F

50µs

, fluorescence intensity at 50 µs

Fj

Fj = fluorescence intensity at J-step (at 2 ms)

Fi

Fi = fluorescence intensity at i-step (at 30 ms)

Fm

Fm = maximal fluorescence intensity

Fv

Fv = Fm - F0 (maximal variable fluorescence)

Vj

Vj = ( Fj - F0 ) / ( Fm - F0 )

Vi

Vi = ( Fi - F0 ) / ( Fm - F0 )

Fm / F0

FV / F0

Fv / Fm

M0 or (dV/dt)0

M0 = TR0 / RC - ET0 / RC = 4 ( F

300

- F0 ) / ( Fm - F0 )

Area

Area between fluorescence curve and Fm (background subtracted)

Fix Area

Area below the fluorescence curve between F

40µs

and F

1s

(background subtracted)

SM

SM = Area / ( Fm - F0 ) (multiple turn-over)

SS

SS = the smallest SM turn-over (single turn-over)

N

N = SM . M0 . ( 1 / VJ ) turn-over number QA

Phi_P0

Phi_P0 = 1 – ( F0 / Fm ) (or Fv / Fm)

Psi_0

Psi_0 = 1 - VJ

Phi_E0

Phi_E0 = ( 1 – ( F0 / F

M

)) . Psi_0

Phi_D0

Phi_D0 = 1 – Phi_P0 – ( F0 / Fm )

Phi_Pav

Phi_Pav = Phi_P0 ( SM / tFm ) t

Fm

= time to reach Fm (in ms)

ABS / RC

ABS / RC = M0 . ( 1 / VJ ) . (1 / Phi_P0 )

TR0 / RC

TR0 / RC = M0 . ( 1 / VJ )

ET0 / RC

ET0 / RC = M0 . ( 1 / VJ ) . Psi_0

DI0 / RC

DI0 / RC = ( ABS / RC ) – ( TR0 / RC )

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Formulas Derived From: R.J. Strasser, A. Srivastava and M. Tsimilli-Michael (2000): The fluorescence transient as a tool to characterize and screen photosynthetic samples. In: Probing Photosynthesis: Mechanism, Regulation and Adaptation (M. Yunus, U. Pathre and P. Mohanty, eds.), Taylor and Francis, UK, Chapter 25, pp 445-483.

7.3.2 NON-PHOTOCHEMICAL QUENCHING (NPQ) PROTOCOL

The NPQ protocol is the most typically used measuring approach to quantify photochemical and non-photochemical quenching. The measurement should be performed with a dark-adapted sample. Thereby, it may not be appropriate under field conditions.

The NPQ protocol starts by giving a measuring light to acquire minimal level of fluorescence F0. A short saturating flash of light is then applied to reduce the plastoquinone pool and measure maximum fluorescence in the dark-adapted state, Fm. After a short dark relaxation, the sample is exposed to actinic irradiance for tens to hundreds of seconds to elicit a transient of the Kautsky effect. Moreover, a sequence of saturating flashes is applied on top of the actinic light to probe the non-photochemical quenching NPQ and effective quantum yield of photosynthesis QY in light adapted state. After exposure to continuous illumination, the relaxation of non-photochemical quenching is determined by means of saturating pulses applied in dark (Fig. 8).

Three NPQ protocols, NPQ1, NPQ2 and NPQ3 are predefined. The protocols differ in the duration of the light exposure and the dark recovery phase, in the number and interval between pulses. See table below:

Phase

Duration

# of pulses

1st pulse

Pulse interval

NPQ1

Light

60 s 5 7 s

12 s

Dark recovery

88 s 3 11 s

26 s

NPQ2

Light

200 s

10

10 s

20 s

Dark recovery

390 s 7 20 s

60 s

NPQ3

Light

200 s

10

11 s

21 s

Dark recovery

60 s 2 20 s

21 s

Tab. 2 NPQ Protocols.

The protocol includes following measured and calculated parameters:

Abbreviation

Explanation

F

0

minimum fluorescence in dark-adapted state

F

m

maximum fluorescence in dark-adapted state, measured during the first saturation flash after dark adaptation

F

p

fluorescence in the peak of fast Kautsky induction

Fm_L, Lss, D, Dss

1

maximum fluorescence

QYmax

2

maximum quantum yield of PSII in dark-adapted state - Fv/F

m

QY_L, Lss, D, Dss

1,3

effective quantum yield of PSII

NPQ_L, Lss, D, Dss

1,4

non-photochemical chlorophyll fluorescence quenching

Qp_L, Lss, D, Dss

1,5

coefficient of photochemical quenching, an estimate of open PSII reaction centers

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1

L - indicates light adapted parameters; D - refers to dark recovery phase after switching of the actinic illumination; n -

represents a sequential number of light phase; ss - steady state

2

Calculated as (Fm – F0) / Fm

3

Calculated as (Fm_Ln – Ft_Ln) / Fm_Ln or of corresponding steady state or dark recovery parameters

4

Calculated as (Fm – Fm_Ln) / Fm_Ln or of corresponding ss, Dn or Dss parameters

5

Calculated as (Fm_Ln – Ft_Ln) / (Fm_Ln – F0_Ln) or of corresponding ss, Dn or Dss parameters

F0_Ln is calculated as F0 / ((Fm – F0) / Fm + F0 / Fm_Ln).

For more details please refer to: Oxborough K., Baker N.R. (1997): Resolving chlorophyll a fluorescence images of photosynthetic efficiency into photochemical and non-photochemical components: calculation of qP and Fv’/Fm’ without measuring F0’. Photosynthesis Research 54: 135-142.

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Fig. 8 NPQ Protocol.

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7.3.3 LIGHT CURVE (LC) PROTOCOL

The protocol called Light Curve (LC) was designed to acquire parameters for construction of Light Response Curve relating the rate of photosynthesis to photon flux density. The method is based on successive measurements of the sample exposed to a stepwise increase of light intensity. The effective quantum yields of photosynthesis are determined under various light intensities of continuous illumination. Measurement is based on pulse modulated fluorometry (PAM).

Several LC protocols are predefined in FP. These differ in number and duration of individual light phases and light intensities.

# of phases

Phase duration

Light intensities [µmol.m-2.s-1]

LC1 6 60s

10; 20; 50; 100; 300; 500

LC2 5 30s

100; 200; 300; 500; 1000

LC3 7 60s

10; 20; 50; 100; 300; 500; 1000

Tab. 3 LC Protocols.

The protocol includes following measured and calculated parameters:

Abbreviation

Explanation

F

0

minimum fluorescence in dark-adapted state

F

m

maximum fluorescence in dark-adapted state

Fm_Ln

‡

maximum fluorescence in light adaptation state

Ft_Ln

‡

instantaneous fluorescence during light adaptation

QYmax*

maximum quantum yield of PSII in dark-adapted state - Fv/Fm

QY_Ln‡**

instantaneous PSII quantum yield induced in light

‡

n represents a sequential number of light phase

*Calculated as (Fm – F0) / Fm

** Calculated as (Fm_Lx – Ft_Lx) / Fm_Lx

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Fig. 9 LC1 Protocol.

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Fig. 10 LC2 Protocol.

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7. 4 MU L TIP L E M EA S UR EME N T

Except single measurement you can perform multiple measurements.

In this mode FluorPen repeats measurements according to parameters, which you have set in Settings > Multi (see Menu tree, page 28)

Multi type – choose your required parameter - Ft, QY, OJIP....

Multi interval – set the time interval between measurements

Multi repeats – set the number of repeated measurements

Use averaging – serves to confirm Repeat and Interval Options for each measurement within Multi Option – select YES or

NO.

• Prepare the sample like for single measurement.

• Go to Measurement > Multi.

• Press SET to confirm.

• Values appears on display after each repeat of measurement and are automatically stored to the device

memory. If you used protocol (OJIP, NPQ, LC) all data you will see after download data (page 46).

Modes of Multiple measurement:

1. FluorPen is connected via USB to computer

The device measures predefined number of repeats and does not swicht off between measurements. Progress of the measurement is displayed in percentage.

2. FluorPen is not connected to the computer

The device measures continuously according to predefined protocol and interval. The multiple measurement is interrupted only by manual switching on of the device. The device turns off between measurements.

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8 CONTROL MENU TREE

The next pages show the structure of the operation scheme, which includes the Main Menu, first-level Sub-Menus and second-level Sub-Menus.

• The blue color represents the Main Menu and its Options.

• The yellow color represents the first-level Sub-Menus and their Options.

• The green color represents the second-level Sub-Menus and their Options.

• Full-line arrows are used for the SET key.

• Dashed-line arrows are used for the MENU key.

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30

31

32

33

34

35

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9 USB CONNECTION

Connect the USB cable with the FluorPen device. Please note that lock in system is used.

Pay attention when connecting the USB cable not to damage the outlet connector on the Pen device. Make sure that you orient the cable correctly prior connecting the inlet with the outlet and the cable is upright towards the device.

To connect FluorPen with your computer please follow steps below in (Fig. 11).

Fig. 11 How to connect FluorPen with PC.

A) Outlet connector on FluorPen device. B) Inlet part on the USB cable. C – E) Position the cable horizontally, plug in the inlet and screw the securing screw. F) Correct connection of the USB cable and Pen device.

Connect the USB cable to a computer. The FluorPen switches ON automatically after connecting the cable to the PC. For USB connection you need to have the USB driver installed in your PC. You find the driver on the installation disk (USB driver folder). If you check the Device Manager in Windows you should see the USB serial port in the device tree. In case of missing driver you may download it from PSI websites. When the driver is installed correctly you should be able to connect to the device in the FluorPen software menu Setup > Device ID.

For more information about FluorPen software see chapter 11.

Monitoring Pen device equipped with Battery Pack has to be connected to PC through Battery Pack.

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10 BLUETOOTH CONNECTION

Before you set up the Bluetooth connection between the FluorPen and your PC, make sure you have these components:

1. Bluetooth enabled PC

The PC with which you connect must have Bluetooth wireless technology, either built-in or through a Bluetooth card. Make sure that the PC's Bluetooth setting is "discoverable" (meaning that it shows up when other devices search for nearby Bluetooth connections). Consult the user guide for your PC or Bluetooth card to learn how to do this.

2. Bluetooth configuration software properly set up on PC

Before you can download files to your PC, you will need to set up the Bluetooth software that came with your PC, or your PC's Bluetooth card. This software varies by manufacturer. Please consult your PC's Bluetooth documentation for more information.

3. Bluetooth must be switched on visible on both devices

To pair the FluorPen with another Bluetooth device, such as a computer, you will need to ensure that Bluetooth is switched on visible on both devices.

10 . 1 B LU E TO O TH PA I RI N G

1. Enabling Bluetooth in the FluorPen

• Switch ON the FluorPen (press and hold the SET key).

• Scroll to the Accessories menu (press the MENU key, then press the SET key).

• Select Bluetooth On (press the MENU key, then press the SET key) to enable Bluetooth.

Keep in mind that the FluorPen turns off automatically after about 8 minutes of no action.

Turning off the device always turns off Bluetooth.

2. Starting Bluetooth Application on Your PC

Be aware that this description is working on Windows 7; some of the steps may be different on your PC.

• Select: Start > Devices and Printers (Fig. 12).

You may also start your Bluetooth application via the Control Panel: Start > Control Panel > Hardware and Sound > Devices and Printers.

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Fig. 12 Start Bluetooth Application.

3. Opening the Add Bluetooth Device Application

• Select: “Add a device” to start searching new Bluetooth device. Be sure that the FluorPen is in discoverable

mode (see step 1).

Fig. 13 Add a device.

4. Selecting the FluorPen

• Select: PSI FluorPen icon.

• Select: Next (Fig. 14).

Fig. 14 Select the FluorPen.

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5. Starting the Pairing Process

This step is different for old and new version of Bluetooth module.

Old version:

Your Bluetooth Pairing Code is: 0000

• Select: “Enter the device’s pairing code”.

• Enter: 0000 (four digits).

• Select: Next ( Fig. 15).

Fig. 15 Pairing process.

New version:

• Select: Yes (Fig. 16). Please note that the FluorPen device does not display the verification number. The

verification code is not important for the BT connection.

• Select: Next.

Fig. 16 Verifying of the BT pairing.

6. Completing the FluorPen Pairing

• Select: Close (Fig. 17).

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Fig. 17 Finishing.

On computer run the program FluorPen 1.1. For more information about FluorPen software see chapter 11.

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11 FLUORPEN SOFTWARE

11 . 1 S OF T WA R E I NS T AL L AT IO N

1. Save the FluorPen software provided on the USB flash disk to your computer and launch the FluorPen program.

2. To connect and recognize your FluorPen device in the FluorPen software proceeds first with the registration of

your FluorPen software (Fig. 18).

• Select: Help > Register

• Enter: your serial registration number.

• Select: OK

Fig. 18 Software registration.

Please note that you will find your serial registration number in the file SN.txt, which is included on the enclosed USB flash disk.

Please remember: it is not possible to download data from the FluorPen device without software registration.

3. Switch on the FluorPen and enable Bluetooth or connect USB cable to the PC.

4. Make sure that your PC and the FluorPen are properly paired (see chapter Error! Reference source not found.

and 10 for complete information on USB and Bluetooth pairing).

5. Select: Setup > Device ID (Ctrl+I). If properly connected, the message “Device: FluorPen” appears in the bottom

part of the screen (Fig. 19).

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Fig. 19 Connecting FluorPen device with Software.

11 . 2 M EN U A N D I CO N E X PL AN A TI O N

11.2.1 MAIN MENU

MENU: File

Load Loads previously saved data files.

Save Saves data to hard disc.

Export Exports data in .txt format.

Export to JSON Exports data in JavaScript Object Notation.

Close Closes the current experiment.

Close All Closes all running experiments.

Exit Exits the program.

MENU: Device

Download Downloads data from the FluorPen to your PC.

Erase Memory Erases data from the FluorPen memory.

Online Control Online control of FP device.

Attach GPS File Used for download data from GPS module (active

only in FluorPen version FP 100 and Monitoring Pen MP 100).

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MENU: Setup

Device ID Detects the connected device.

Update Firmware Used for firmware updates.

Settings Used for modification of the program

settings.

MENU: Help

About Offers basic information about the program.

Register Used for the FluorPen software registration.

Icon Explanation:

11.2.2 MENU SETTINGS

MENU > Setup > Settings

After Download – Memory Erase

If the box is checked the FluorPen memory is erased after each data download.

Data – Inverted

If the box is checked the polarity of data is inverted, e.g., multiplied by -1. This feature can be helpful for a certain type of experiment when the measured data are undesirably interpreted as negative values.

Data – Add to opened

If the box is checked the downloaded data are added to that of the current opened experiment.

Download

Downloads data from the FluorPen to PC.

Load

Loads (opens) previously saved data files.

Save

Saves data to hard disc.

Export

Exports data in .txt format.

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Data – GPS Visible

This option is active only in older FluorPen version FP 100 and Monitoring Pen MP 100. In new versions of FP 110 the GPS data are automatically downloaded and paired with protocol measurements.

Graf – Single

If the box is checked all measured data are visualized in one graph, i.e., the value of each new measurement is added to the currently used graph window.

If the box is not checked a new graph is opened for every new measurement.

11.2.3 MENU ONLINE CONTROL

This function can be used for Online Control your FluorPen device after connection with your PC.

• Select: Menu > Device > Online Control

Online Control – Switches

Sound On/Off - choose On/Off for device beeping.

Multi use averaging (YES/NO) – serves to confirm Repeat (number of repetitions) and Interval (time between measurements) Options for each measurement within Multi Option – select YES or NO.

Online Control – Values

You can change settings of Actinic, Super or Flash Pulse light. You can also set Averaging of Ft, OJIP, NPQ, LC measurements. You can set time between measurements (Interval) and also you can set number of Multiprotocol measurements (1-1000), type of Multiprotocol or time between measurements.

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Online Control – Time

Set the FluorPen time a date. You can also synchronize time of FluorPen device with computer time.

Online Control – Protocols

Choose the protocol for measuring. FluorPen device start to measure. After measuring you can download data. Measuring of PAR is active only in the PAR-FluorPen FP 110.

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11 . 3 D AT A T R AN S FE R A N D VI S UA L I ZA T IO N

1. Perform a measurement with your FluorPen. In Fig. 20 we did an OJIP and NPQ measurement.

2. Click the Download icon or select Device > Download.

3. The Data table appears.

Fig. 20 Example of Data Transfer and Visualization.

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4. To visualize measurement in the graphic mode, click the Graph field in the bottom bar.

5. The colored Graph of measured data appears (Fig. 21).

Fig. 21 Graphic visualization of experiment.

6. For export press File > Export or Export icon, select what you want to export (Ft, QY, OJIP…) (Fig. 22).

Selected only – exports only one measurement that is selected by mouse, otherwise it will export everything. Source data – exports raw data, in case of OJIP: points of the curve. Description – exports the data description if any. Computed values – export calculated data, in case of OJIP: F0, Fi, Fj...

Fig. 22 Export data.

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11 . 4 F IR M WA R E U PD ATE

All data in the FluorPen memory are erased during the firmware update!

Before starting any firmware update, export all your data from the FluorPen memory into your computer!

1. Starting Update

• Select: Setup > Update Firmware From File (Fig. 23).

Fig. 23 Update Firmware.

2. Warning

• Select: OK to start update (Fig. 24).

3. Selecting .bxn file

• Find: Binary file (with the extension .bxn) (Fig. 25).

• Select: Open.

Fig. 24 Warning.

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Fig. 25 Select .bxn file.

4. Finishing Upload

• Select: OK to start uploading of the update (Fig. 26).

Fig. 26 Data loss warning.

• The bottom bar indicates the upload progress (Fig. 27).

Fig. 27 Upload progress.

• Press: OK to finish upload (Fig. 28).

Fig. 28 Finish upload.

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12 GPS MODULE

FluorPen device has integrated GPS module which can be turned on during the measurement and the GPS coordinated will be added to the downloaded data.

For proper GPS reading, the time in your FluorPen and in your computer must be synchronized. Preset time and time zone must correspond to GPS time (time zone) in your location.

12 . 1 G PS / F LU O RP E N O PE R AT I ON

1. Check the time setting in FluorPen device: Settings > Date & Time

2. Switch the GPS module on:

• Select: Accessories > GPS

• Press SET to confirm.

• Wait until the GPS position is found – “Starting GPS”.

• The GPS module is ready when the icon in upper panel change – see on Fig. 29.

Fig. 29 GPS icons.

3. Go to Measurement and choose required protocol.

The device may need a clear view of the sky to acquire satellite signal.

Keep in mind that the FluorPen turns off automatically after about 8 minutes of no action.

Turning off the FluorPen always turns off GPS module.

For prompt determination of the coordinates use the option Accessories > GPS > Location.

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12 . 2 D AT A D O WN LOA D

1. Enabling Communication:

• Switch on the computer and the FluorPen device. Set your computer to FluorPen communication: enable

Bluetooth or connect to USB port.

2. Downloading Data from the FluorPen

• Start FluorPen program.

• Connect FluorPen device: Setup > Device ID (Ctrl+I)

• Download measured data from the FluorPen to your PC. Data measured with activated GPS module are

downloaded with GPS coordinates (Fig. 30).

Fig. 30 GPS coordinates.

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13 WARRANTY TERMS AND CONDITIONS

• This Limited Warranty applies only to the FluorPen device. It is valid for one year from the date of shipment.

• If at any time within this warranty period the instrument does not function as warranted, return it and the

manufacturer will repair or replace it at no charge. The customer is responsible for shipping and insurance charges (for the full product value) to PSI. The manufacturer is responsible for shipping and insurance on return of the instrument to the customer.

• No warranty will apply to any instrument that has been (i) modified, altered, or repaired by persons

unauthorized by the manufacturer; (ii) subjected to misuse, negligence, or accident; (iii) connected, installed, adjusted, or used otherwise than in accordance with the instructions supplied by the manufacturer.

• The warranty is return-to-base only, and does not include on-site repair charges such as labor, travel, or other

expenses associated with the repair or installation of replacement parts at the customer's site.

• The manufacturer repairs or replaces faulty instruments as quickly as possible; the maximum time is one month.

• The manufacturer will keep spare parts or their adequate substitutes for a period of at least five years.

• Returned instruments must be packaged sufficiently so as not to assume any transit damage. If damage is

caused due to insufficient packaging, the instrument will be treated as an out-of-warranty repair and charged as such.

• PSI also offers out-of-warranty repairs. These are usually returned to the customer on a cash-on-delivery basis.

• Wear & Tear Items (such as sealing, tubing, padding, etc.) are excluded from this warranty. The term Wear &

Tear denotes the damage that naturally and inevitably occurs as a result of normal use or aging even when an item is used competently and with care and proper maintenance.

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14 TROUBLESHOOTING AND CUSTOMER SUPPORT

In case of troubles and for customer support, please, visit FAQ on our websites, write to support@psi.cz or contact your local distributor.

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15 APPENDIX

15 . 1 B AT T ER Y P A CK FO R M ON I TO R IN G P E N

Battery pack serves as external battery source for Monitoring pen devices. The external battery ensures powering of

devices during long-term experiments.

15.1.1 STANDARD BATTERY PACK

Standard battery pack (Fig. 31) is intended for operation within temperature range from +10 ºC to +40 ºC. The operating time is up to 2 years (QY measurement every 1 hour). The pack includes battery case with rechargeable sealed lead acid battery (12Ah), charger, two types of cables (serial and device) and serial convertor.

Fig. 31 Standard Battery Pack.

Connectors of Battery Pack (Fig. 32):

Device (as FluorPen in Fig. 32)

Serves for connection between Monitoring Pen and the battery. This connection is necessary for Pen operation and data download.

Serial

The connector enables communication between Monitoring Pen and PC.

Charger

The connector is serving for charging the battery.

Please note that Monitoring Pen devices equipped with battery pack do not have internal battery, therefore it is not possible to use them separately without the battery pack.

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Fig. 32 Connectors of Standard Battery Pack.

Battery exchange:

1. Unscrew 4 screws and remove the battery pack cover (Fig. 33a).

2. Disconnect the internal battery from the cables.

3. Place new battery into the pack, connect it with the cables – red cable with red marked connector and black

cable with black marked connector (Fig. 33b, c). Put back the cover.

Fig. 33 Battery exchange of Standard Battery Pack.

15.1.2 EXTENDED TEMPERATURE RANGE BATTERY PACK

Extended temperature range battery pack (Fig. 34) is intended for operation within temperature range from -40 ºC to +60 ºC. Operating time is up to 2 years (QY measurement every 1 hour). The pack includes battery case with nonrechargeable Li-SOCl2 battery (5.5Ah), two types of cables (serial and device) and serial convertor.

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Fig. 34 Monitoring Pen MP 100-A charged via Extended Temperature Range Battery Pack.

Connectors of Battery Pack:

Device

Serves for connection between Monitoring Pen and the battery. This connection is necessary for Pen operation and data download.

Serial

The connector enables communication between Monitoring Pen and PC.

Battery exchange:

1. Unscrew 4 screws and remove the battery pack cover (Fig. 35 a, b).

2. Disconnect the internal battery from the cable (Fig. 35 c).

3. Place new battery into the pack, connect it with the cable and put back the cover.

Extended temperature range battery pack cannot be recharged. Spare battery is offered as additional accessory.

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Fig. 35 Battery exchange of Extended Temperature Range Battery Pack.

PSI FluorPen FP 110, PAR-FluorPen FP 110, Monitoring Pen MP 100 Manual And User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual