FMS-300

Pulse Amplitude Modulated (PAM) chlorophyll fluorometer for teaching & research

  • Versatile functionality
  • Fast & slow fluorescence data for every saturating flash event
  • Comprehensive parameter display
  • Developed for high performance
  • Streamlined data capture
  • Intuitive but powerful software
  • Data visualisation & analysis
  • Measuring light & actinic/saturating LED colours
 

Welcome to FMS-300

FMS-300 is a state-of-the-art, Pulse Amplitude Modulated (PAM) chlorophyll fluorometer combining the usability of a teaching system with the power and functionality to provide high-level research-grade data. Key features of the FMS-300 are as follows:

  • Versatile functionality:
    Suitable for both teaching and research applications. FMS-300 can demonstrate complex concepts, conduct experiments, collect data, analyse results, and facilitate collaborative learning or research projects.

  • Fast & slow fluorescence data:
    Presents fast and slow fluorescence data with equal emphasis. Fast fluorescence kinetics are captured during every saturating pulse for both dark- and light-adapted sample states, presenting opportunities for novel research.

  • Comprehensive parameter display:
    Parameter sets relevant to the selected routine are calculated for each saturating pulse. For dark-adapted samples, OJIP parameters in addition to parameters such as Fv/Fm are calculated. Light-adapted parameters including ETR, ΦPSII, Lake and Puddle models for non-photochemical quenching, and both a calculated Fo’ and Fo’ measured under far-red light are calculated where appropriate.

  • Developed for high performance:
    A measuring pulse width of 400 ns from a dedicated LED allows greater intensities to be implemented yielding a strong, low-noise signal. Measuring pulse intensity can be defined between *0.001 – 0.1 µmol m-2 s-1. Actinic and saturating pulse intensities of up to 3,000 and **20,000 µmol m-2 s-1 respectively can be delivered at the sample surface.

  • Streamlined data capture:
    FluoroControl software provides 6 pre-set experimental routines, which can be further configured, if required. This enables rapid learning and efficient, hassle-free data acquisition.

  • Intuitive software with simplified control set:
    Developed from the ground up with simplicity in mind. A user-friendly interface with minimal learning curve allows easy navigation of the different functions and options.

  • Data visualisation & analysis:
    The measuring LED performance means that 100% raw data is presented with no requirement to damp or average. Robust data visualisation, analysis and export tools allow users to interpret and present findings effectively. Valuable for both teaching complex concepts and conducting in-depth analysis in research applications.

  • Measuring light & actinic/saturating LED colours:
    4 variants with different pairings of LED colour. Supplied with either blue/blue (455 nm), red/red (624 nm), blue/white or red/white measuring/actinic and saturating LEDs. All variants contain a far-red LED (730 nm) for determination of Fo’ during the quenching analysis routine.

FMS-300 Overview

FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications

FMS-300 is a state-of-the-art, Pulse Amplitude Modulated (PAM) chlorophyll fluorometer combining the usability of a teaching system with the power and functionality to provide high-level research-grade data. A wide range of accessory components allow experiments to be conducted on many different sample types.

Newcomers to the technique are quickly able to acquire and analyse data, making FMS-300 an ideal system for teaching chlorophyll fluorescence. Yet it is also a highly capable research instrument offering flexibility, functionality, and data acquisition of exceptional quality. Extensive features and capabilities allow the system to be used to demonstrate complex concepts, conduct experiments, collect data, analyse results, and facilitate collaborative learning or research projects.

FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications

Primarily a laboratory-based system, FMS-300 can extend to greenhouse and field applications when coupled with an appropriate portable power source.

The enviable signal quality is achieved via ultra-short measuring pulses with a standard frequency of 10 Hz (up to 100 kHz during fast fluorescence capture). At just 400 ns per pulse, FMS-300 can emit high-intensity measuring pulses with user-defined average intensities up to 0.1 μmol m-2 s-1. This combination of high-intensity and ultra-short pulse width produces a strong fluorescence signal with no requirement for signal damping or data averaging. The user is presented with 100% raw instrument data.

FluoroControl Software

A free copy of FluoroControl software and a set of example data files is available on request with no purchase necessary. Please click on the Product Enquiry tab and complete the short form to request a link to download the software.

FMS-300 is supplied with a sophisticated, yet user-friendly software package. FluoroControl controls all aspects of instrument operation and has been developed with simplicity of use at its core. An intuitive, tab-based interface ensures a shallow learning curve allowing users to begin measurements quickly.

FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications

Configuration of the FMS-300 hardware is achieved from a single settings screen. 6 configurable measurement routines, which cover all conventional fluorescence protocols, are used to acquire data.

A comprehensive set of measured and calculated parameters are presented with fast fluorescence (OJIP) data captured for every saturating pulse event in both light- and dark-adapted states. Measured data can be easily exported to CSV format for further analysis in external software.

Fluorescence measurement routines

FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications
FMS-300 is operated via the use of 6 pre-set typical experimental routines, configurable if required. This enables rapid learning and efficient, hassle-free data acquisition.

Each routine’s protocol consists of saturating pulse events with periods of measurement in the dark or under actinic or ambient light conditions. This allows fluorescence yield measurements under a wide range of environmental conditions. FMS-300 captures extremely high-resolution fast fluorescence kinetics during each saturating pulse event for both light- and dark-adapted sample states.

Routine 1: Maximum QY
This routine provides a measurement of the Fv/Fm parameter; the maximum quantum yield of PSII photochemistry, and a full suite of OJIP (JIP-Test, Strasser et al., 2004) analysis parameters, performed on a dark-adapted sample.
Parameters recorded: Fv/Fm, Fo, Fm, Fv, TFm, Area, F20µs, FL, FK, FJ, FI, Fv/Fo, Fo/Fm, VJ, VI, RC/ABS, dVg/dto, N, Sm, Sm/TFm, Mo, φPo, φEo, φRo, ψEo, δRo, PIabs, PItotal, DFabs, DFtotal, ABS/RC, TRo/RC, ETo/RC, REo/RC, DIo/RC, ABS/CSo, TRo/CSo, ETo/CSo, REo/CSo, DIo/CSo, ABS/CSm, TRo/CSm, ETo/CSm, REo/CSm, DIo/CSm, kN, kP.

Routine 2: Effective QY
Provides a measurement of the ΦPSII (or Y[II]) parameter from a light-adapted sample at steady-state photosynthetic activity. ΦPSII is an estimation of the effective quantum yield of PSII photochemistry.
Parameters recorded: ΦPSII, Fv’/Fm’, F, Fm’, Fq’, TFm’, Fo’(measured), Fo’(calculated), Fv’, F20µs’, FL’, FK’, FJ’, FI’, VJ’, VI’.

Routine 3: Quenching
Performs a full quenching analysis according to the protocol defined in Baker (2008), with estimation of non-photochemical quenching. Parameters for both Puddle and Lake models (Kramer 2004, Hendrickson 2004) are presented along with other data relating to light- and dark-adapted sample states.
Parameters recorded: All parameters relevant to light- and dark-adapted measurements plus:
Puddle Model: NPQ, qN, qP
Lake Model (Kramer): ΦPSII, Y(NPQ), Y(NO), qL
Lake Model (Hendrickson): ΦPSII, Y(NPQ), NPQ, Y(NO).

Routine 4: Light Response Curve
Analyses the electron transport rate (ETR) parameter for a series of short periods of actinic illumination with increasing intensity. The software plots ETR vs PPFD and calculates parameters including maximum ETR (ETRmax) and the minimum saturating irradiance (Ek). Actinic step durations can be lengthened to allow steady-state photosynthesis to be achieved for each light level.
Parameters recorded: All parameters relevant to light-adapted measurements plus:
ETR, JNPQ, PAR, α, β, ETRmax, Ek.

Routine 5: Multi-Maximum QY
Allows successive measurements of dark-adapted Fv/Fm / JIP-Test measurements to be made. Allows comparison of several different samples, replication or screening of, for example, multiple phenotypes.
Parameters recorded: As routine 1 parameters.

Routine 6: Multi-Effective QY
Allows successive measurements of light-adapted ΦPSII measurements to be made for comparison, replication or screening applications.
Parameters recorded: As routine 2 parameters.

A free copy of FluoroControl software and a set of example data files is available on request with no purchase necessary. Please click on the Product Enquiry tab and complete the short form to request a link to download the software.

Fast fluorescence kinetics for all saturating pulse events

FluoroControl software presents fast and slow fluorescence data with equal emphasis. Fast fluorescence kinetics are measured and plotted for every saturating pulse event for both dark- and light-adapted sample states. This ability, not typically found in commercial fluorometers, presents opportunities for novel research. For example, studying the changes in the fast fluorescence kinetics during a quenching routine can show PSII redox activity as PSI and NPQ components begin to operate. These processes happen when a dark-adapted sample becomes fully light-adapted under constant actinic illumination. Later in the quenching protocol, the same redox states can be studied in reverse as the fully light-adapted sample becomes re-dark-adapted as NPQ components begin to relax in the dark.

FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications

Sampling frequency during slow kinetics is 10 Hz (1 sample every 100 ms) which increases to 100 kHz (1 sample every 10 µs) during fast-fluorescence capture. This rapid sampling rate is essential in accurately capturing the early phases of the Kautsky Effect (OJIP) which provides information regarding the redox state of the primary electron acceptors of PSII.

A free copy of FluoroControl software and a set of example data files is available on request with no purchase necessary. Please click on the Product Enquiry tab and complete the short form to request a link to download the software.

Dark-adaptation leafclips

These should be used when dark-adaptation of the sample (or measurement in the absence of light), is required before or during the experiment. The leafclips can be attached to samples independently prior to measurements to facilitate adequate dark-adaptation time for the sample being measured without hindering throughput.

Dark-adaptation leafclips can be used for a wide variety of different samples.

Dark-adaptation leafclips for FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications

Dark-adaptation leafclips are constructed from 3D printed nylon. To dark-adapt the sample, a stainless steel shutter plate in the upper section of the clip is slid forward, covering the sample, and effectively preventing ambient light from reaching the leaf. During measurements, the sampling end of the fibre-optic cable is inserted into the fibre receptor on the upper section of the leafclip and securely fastened using the thumb-screw. Before taking measurements, the shutter is retracted to the open position, exposing the dark-adapted sample to the fibre-optic light source.

The sample rests on a foam pad whilst in the clip in order to minimise damage to the structure of the sample.

During dark adaptation, all the reaction centres are fully oxidised and available for photochemistry and any chlorophyll fluorescence yield is quenched. This process takes a variable amount of time and depends upon plant species, light history prior to the dark transition and whether or not the plant is stressed. Typically, 15 – 20 minutes may be required to dark adapt effectively. In order to reduce waiting time before measurement, a number of leaves may be dark adapted simultaneously using several leafclips.

PTL-100 leafclip

FMS-300 is supplied as standard with an open-faced leafclip, the PTL-100, for measurements of chlorophyll fluorescence under ambient/artificial light conditions. It can be used for routines when measurement of samples in a light-adapted state is required.

PTL-100 Open leafclip with PAR and temperature sensors for FMS-300

The PTL-100 consists of a 3D printed nylon body with a front-mounted PAR sensor. A leaf temperature sensor is mounted within the underside of the leafclip. The fibre-optic is inserted through a retaining collar and secured in place with a thumbscrew with the open end of the fibre-optic positioned at 60° to the plane of the sample and in close proximity to maximise both sample illumination intensity and detection of the fluorescence signal. The PTL-100 is fitted with an adapter at the rear of the leafclip which allows connection to the fibre-optic support arm. The sample is held securely in position by a magnetic “puck” that is placed underneath the leaf.

Fibre-optic cable support arm and clamp

The FMS-300 is supplied as standard with an articulated support arm and clamp. These items assist with convenient, effective and secure positioning of the fibre-optic during measurements. They can be used in conjunction with the FMS-300 branded retort stand (available as an optional accessory) or with standard laboratory retort stands.

FMS-300 fibre-optic support arm: Pulse-modulated chlorophyll fluorometer for teaching & research applications

At one end of the arm is a 9.5mm male thread which connects to the 9.5mm female thread on the rear of the clamp. At the other end, the arm is fitted with an adapter that allows the FMS-300 fibre-optic cable to be held securely in place during measurements.

The jaws of the clamp are opened and closed using the handle on the side of the clamp. The jaws will securely grip on to cylindrical items from 13 – 35mm in diameter and support loads of up to 4kg. With the support arm and clamp securely attached to a retort stand, the arm may be adjusted through a wide range of movement to conveniently position the sample end of the fibre-optic cable as required.

A choice of 4 LED colour options

The maximum intensity of the measuring LED in the B and BW variants is up to 0.1 µmol m-2 s-1. In the R and RW variants, maximum intensity of the measuring LED is up to 0.058 µmol m-2 s-1. Whilst all variants offer exceptional signal to noise qualities, for measurements on most eukaryotic algae or higher plants, the FMS-300-B and BW variants provide the strongest fluorescence response. It is important to understand the absorption characteristics and photochemical requirements of the intended sample when choosing the most suitable FMS-300 variant.

Variant Measuring LED Actinic &
Saturating LED
Far-red LED Fluorescence
Detector
FMS-300-B Blue – 455 nm (FWHM 27 nm) 730 nm
(FWHM 20 nm)
> 645 nm
FMS-300-R Red – 624 nm (FWHM 18 nm) > 680 nm
FMS-300-BW Blue – 455 nm
(FWHM 27 nm)
Ultra-white
FMS-300-RW Red – 624 nm
(FWHM 18 nm)

Blue light is maximally absorbed at the upper surface and does not penetrate deeply into the leaf (Ramos & Lagorio, 2004). Red light penetrates deeper into the leaf inducing fluorescence over the entire cross section although a significant proportion of fluorescence is reabsorbed (Gitelson et al. 1999) resulting in an overall decrease in fluorescence intensity. Blue-induced fluorescence is emitted from the surface of the leaf only with very little re-absorption resulting in a higher fluorescence intensity and a strong, low noise fluorometer signal (Ramos & Lagorio, 2004).

The use of blue excitation light wouldn’t be suitable for some experiments. Cyanobacteria contain specialised antennae known as phycobilisomes (PBSs) which are not thought to absorb blue light. Therefore, cyanobacteria have a reduced photosynthetic efficiency under blue light (Luimstra et al. 2018) and the use of a red excitation light would be applicable.

The emission characteristics of each LED relative to the absorption characteristics of chlorophyll a are shown in Figure 1. Figure 2 shows the fluorescence detection characteristics relative to the chlorophyll fluorescence emission spectrum for each variant of FMS-300.

FMS-300-B is fitted with a different fluorescence detector filter to the other variants, allowing fluorescence emission above 645 nm to be captured (Figure 2). This provides an extremely low-noise fluorescence signal since it transmits a significant proportion of the 685 nm emission peak. All other variants detect fluorescence above 680 nm (Figure 2).

Figure 1.

Emission characteristics of FMS-300 LEDs relative to the chlorophyll a absorbance spectrum.

Emission characteristics of FMS-300 LEDs relative to the chlorophyll a absorbance spectrum.

Figure 2.

Detector characteristics of FMS-300 variants relative to the chlorophyll a emission spectrum.

Detector characteristics of FMS-300 variants relative to the chlorophyll a emission spectrum.

References

Ramos, M.E., Lagorio, M.G.
True fluorescence spectra of leaves.
Photochem Photobiol Sci 3, 1063–1066 (2004).
Gitelson, Anatoly A., Claus Buschmann, and Hartmut K. Lichtenthaler.
The chlorophyll fluorescence ratio F735/F700 as an accurate measure of the chlorophyll content in plants.
Remote sensing of environment 69.3 (1999): 296-302.
Luimstra, Veerle M., et al.
Blue light reduces photosynthetic efficiency of cyanobacteria through an imbalance between photosystems I and II.
Photosynthesis Research 138 (2018): 177-189.

FMS-300 optional accessory items

A range of accessory components extend the functionality of FMS-300 for different sample types. Hansatech Instruments offer 3 different light guides made from optical grade, highly polished borosilicate glass. These connect to the fibre-optic cable in place of a standard leafclip, to allow measurements in a wide range of specialised applications.

FMS-300 modulated fluorometer light guides

The 8 x 100mm hexagonal light guide is suited to e.g. cacti and succulents or insertion into water tanks for measurements on e.g. macroalgae, coral. The light guide is protected by a sleeve and laminar samples are held in place with a magnetic puck.

For measuring smaller samples such as Arabidopsis, needles and lichens, two tapered light guides are available. The tapered form of these light guides focuses the light from FMS-300 and increases the maximum saturating pulse intensities. The two light guides have dimensions at the sample end of 4 x 4 mm and 2.5 x 2.5 mm providing up to 60,000 and 90,000 µmol m-2 s-1 respectively. The tapered light guides are also suitable for insertion into liquids for measurements on samples such as seagrass.

FMS-300 modulated fluorometer light guides

For measurements of liquid samples such as algae, a chamber designed to accept 4.5 ml spectrophotometer cuvettes is available. Liquid sample chambers are fitted with a swivel-action lid to enable samples within the chamber to be dark-adapted.

A lateral optical port allows the FMS-300 fibre-optic cable to be inserted into the chamber so that the tip of the fibre-optic is in direct contact with the outer wall of the sample cuvette. Samples with a minimum optical density (O.D.) of 0.1 can be measured with both slow and fast fluorescence kinetics presented following every saturating pulse event. Using multiple chambers means that multiple samples can be dark-adapted simultaneously prior to measurement. Sample chambers are compatible with the Hansatech Instruments magnetic stirrer or existing magnetic stirrer plates so that samples can be mixed during the experiment.

FMS-300: Pulse-modulated chlorophyll fluorometer for teaching & research applications

To complement the supplied fibre-optic support arm, a solid oak-base retort stand with laser-etched Hansatech Instruments and FMS-300 badging is available.

FMS-300/RS: Branded solid English Oak retort stand.

System Components

  • FMS-300 control unit.
    Options for LED colours to be specified at point of order:
    • FMS-300-B: blue measuring, actinic and saturating LED
    • FMS-300-R: red measuring, actinic and saturating LED
    • FMS-300-BW: blue measuring with white actinic and saturating LED
    • FMS-300-RW: red measuring with white actinic and saturating LED
  • FMS/SFO fibre optic cable
  • PTL-100 PAR temperature leafclip
  • FMS/DLC pack of 10 dark-adaptation leafclips
  • Fibre optic cable support arm and clamp
  • Power supply
  • USB connection cable
  • Protective hard case

Control unit

All light intensity specifications refer to incident illumination at the sample surface.

Measuring LED:
Pulse width: 400 nanoseconds.
Pulse frequency: Slow kinetics: 10 Hz.
Fast kinetics: Semi-log frequencies from 10 Hz-100 kHz.
Pulse intensity: Up to 0.1 µmol m-2 s-1 in FMS-300-B/BW.
Up to 0.058 µmol m-2 s-1 in FMS-300-R/RW.
Adjustable in 0.001 µmol m-2 s-1 increments.
With light guide, FMS/LG2: >0.25 µmol m-2 s-1.
With light guide, FMS/LG3: >0.4 µmol m-2 s-1.
Measuring LED colour: FMS-300-B/BW: Blue 455 nm (FWHM 27 nm).
FMS-300-R/RW: Red 624 nm (FWHM 18 nm).
Actinic/saturating pulse LED colour: FMS-300-B/BW: Blue 455 nm (FWHM 27 nm).
FMS-300-R/RW: Red 624 nm (FWHM 18 nm).
Max. actinic intensity: Up to 3,000 µmol m-2 s-1. Adjustable in 1 µmol m-2 s-1 increments.
With light guide, FMS/LG2: >8,000 µmol m-2 s-1.
With light guide, FMS/LG3: >12,000 µmol m-2 s-1.
Max. saturating pulse intensity: >20,000 µmol m-2 s-1. Adjustable in 1 µmol m-2 s-1 increments.
With light guide, FMS/LG2: Up to 60,000 µmol m-2 s-1.
With light guide, FMS/LG3: Up to 90,000 µmol m-2 s-1.
Far-red LED: 730 nm (FWHM 20 nm).
Far-red max. intensity: Up to 20 µmol m-2 s-1.
With light guide, FMS/LG2: up to 60 µmol m-2 s-1.
With light guide, FMS/LG3: up to 90 µmol m-2 s-1.
Detector: PIN photodiode, rapid peak-pulse tracking.
Detection range: FMS-300-B: >645 nm.
FMS-300-R/BW/RW: >680 nm.
Electronics:
Dual processors: ARM 32-bit microcontroller running real-time operating system.
PIC 8-bit microcontroller for dedicated measuring LED sampling.
Fibre-optic cable: Length 1 m, statistically randomised fibre bundles.
7mm optical diameter at sample end.
Connections: Optical connection for fibre-optic cable.
12 V DC socket.
USB-C socket.
MiniDIN socket for PTL-100.
Enclosure: Shielded aluminium enclosure.
Dimensions: 150 (l) x 150 (w) x 85 mm (d).
Weight 770g.
Communications: USB 2.0. Cable type A – C.
Operating conditions: 0°C-40°C. Non-condensing humidity.
Power supply: 12 V DC mains power.
Power consumption: 5.6-7.2 W. Max. 50 W.
Display: 4 line x 20 character blue LCD display.

 

Leafclips & accessories

FMS/DLC: Nylon 3D printed, 7 mm sample aperture, sliding shutter blade, 60° angle between fibre-optic & sample.
PTL-100: Nylon 3D printed, leaf temp. & PAR (400-700 nm) sensor. Electrical connection to FMS-300. Open faced, 60° angle between fibre-optic & sample.
Manfrotto support arm: Mini Variable Friction Arm. 0.44kg,
length: 12-24 cm, max. payload: 3 kg.
Manfrotto clamp: Nano Clamp. 0.097 kg. clamp range 13-35mm, max. payload: 4 kg.
Transport case: 464 (l) x 366 (w) x 176 (d) mm. Weight: ~3 kg.

Software requirements

Operating System: Windows 10 or newer.

To send an enquiry about FMS-300, please complete and submit the form below. If you choose to receive a copy of the FMS-300 FluoroControl software, a time-limited download link will be emailed to you once we have received the request.

Downloads for FMS-300

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66 downloads 4.4 MB

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64 downloads 7.4Mb

A free copy of FluoroControl software and a set of example data files is available on request with no purchase necessary. Please click on the Product Enquiry tab and complete the short form to request a link to download the software.

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