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FS5 Spectrofluorometer

  • Multiple detector ports in one integrated instrument, measure up to 1650 nm and fluorescence lifetimes down to 25 ps, in one instrument
  • >6,000:1 Water Raman SNR, high sensitivity allows for detection of very weak fluorescence signals
  • Three detectors as standard, reference detector to correct for light source fluctuations, transmission detector for absorbance, single-photon counting (SPC) detector for ultimate sensitivity
  • Plug & Play sample modules, automatic sample module recognition and initialisation saves the user time and effort. Wide range of modules available for varied sample analysis
FS5 Spectrofluorometer

The FS5 is designed to meet the highest measurement specifications in the research and analytical markets. To achieve optimal sensitivity, resolution, and acquisition speed, we select only the optimal performance photomultipliers in temperature stabilised housings, and utilise an optical design that includes plane, spherical, toroidal, ellipsoidal mirrors and optimised mirror coatings; the FS5 is truly unrivalled in its spectral performance and sensitivity.

Not only does this superior design guarantee the highest sensitivity and broadest wavelength coverage, but it has also been optimised for a small bright focus at the sample position. This is important for measuring small sample volumes and benefits many sample holder attachments like a plate reader, fibre launch optics mounting, titrators and sample positioners.

The standard FS5 has a number of upgrade routes that are unique in its class, including spectral range extension up to 1650 nm, Fluorescence and Phosphorescence Lifetime measurement capabilities, polarisation / anisotropy and much more. Please see the upgrades tab for more information.

 

 Specification
OpticsAll-reflective for a wavelength independent focus with high brightness (small focus) at the sample
Source150 W CW Ozone-free xenon arc lamp
MonochromatorsCzerny-Turner design with plane gratings for accurate focus at all wavelength and minimum stray light
Spectral Coverage -– Excitation230 nm -– 1000 nm
Spectral Coverage -– Emission230 nm -– 870 nm
Filter WheelsFully automated; included in both the excitation and emission monochromators
Bandpass –- Excitation/Emission0* to 30 nm, continuously adjustable *resolution limit of 0.3 nm
Wavelength Accuracy –- Excitation/Emission± 0.5 nm
Scan Speed –- Excitation/Emission100 nm/s
Integration Time1 ms –- 200 s
Emission DetectorPhotomultiplier R928P, spectral coverage 200 nm -– 870 nm, cooled and stabilsied
Reference DetectorUV enhanced silicon photodiode
Transmission DetectorUV enhanced silicon photodiode
Water Raman Signal>400,000 cps at 397 nm emission, excitation 350 nm, 5 nm bandpass, 1 s integration time
Signal-to-Noise Ratio of Water Raman SignalSNR SQRT >6000:1
Dimensions104 cm (w) x 59 cm (d) x 32 cm (h)
Weight55 kg

Fluoracle® is the operating software for the FS5 Spectrofluorometer. It controls all FS5 steady state and time-resolved spectrometer features with a straightforward design concept: focus on all modern photoluminescence spectroscopy applications and provide a user-friendly interface with ‘ready to publish’ outputs. Whether you select a basic spectral scanning version of FS5, or you go for an advanced version that includes TCSPC lifetime measurements or integrating sphere measurements, the all-inclusive software will provide instrument options automatically, from data acquisition to analysis and presentation.

Upgrade to FAST Software for advanced fluorescence lifetime analysis.


Measurement Examples

Emission Scans – with temperature dependence

Rhodamine B, unlike other Rhodamine derivatives, has a chemical structure that is not entirely rigid. Consequently, the diethylamino groups interact with the solvent’s temperature, affecting the molecule’s excited-state population and dynamics through torsional motion. This causes the fluorescence intensity to have a strong dependence on the sample temperature, which can be seen below as measured with the TE cooled sample holder.

RhodamineB

Rhodamine B in water, OD at 525 nm = 0.1 Spectral Band width: 2.5 nm, integration time per point: 0.1 s Temp Accuracy Band: 0.5ºC, Temp Stabilisation Time: 10 min

Excitation Scans – with pH dependence

Fluorescence excitation spectra are more selective than absorption spectra, as they reveal, by virtue of the selected emission wavelength, where the molecule can absorb photons to prodcue a particular emitting species. Accurate excitation spectra require a sensitive instrument, as the concentration of the sample must be kept low to avoid inner filter effects, and require reliable spectral correction to ensure proper spectral representation.

excitation

Fluorescein in water, with pH 2 – 7 adjustments. Spectral Band width: 1.5 nm, integration time: 0.1 s pH adjusted between pH= 2 (blue shifted spectrum) and pH = 7 (spectrum of maximum intensity)

Synchronous Scans – with concentration dependence

In synchronous spectral scans, the excitation and emission monochromators scan at the same time with a fixed wavelength offset, typically from 0 – 20 nm of offset pending the application. For dilute mixtures, this type of scan is used to identify species with a strong overlap between absorption and emission. Synchronous scans, together with the integrating sphere attachment, can also be used to measure the transmission/reflectance/absorption spectra of strongly scattering powders.

Concentration dependence Concentration dependence

YAG: Ce powder, diluted with BaSO4 to study the effect of re-absorption/emission, concentration change from 100% down to 20%. The software wizard is used to calculate the Absorbance (left) from the raw data of synchronous scans (right).

Absorption Scans / Kinetic Scans

The FS5 can record the time course of a fluorescence signal, and, at the same time, record the signal transmitted through the sample. This enables experiments to be performed with chemically or biologically unstable samples, or with samples where very small changes need to be measured very accurately. The transmission detector is standard in the FS5.

Absorption

Caspase Assay, fluorescence time course recorded for a 100% enzyme addition (blue) and a 0% enzyme control (red). Peptide cleavage is recorded by an organic dye excited at 400 nm, emitting at 460 nm.

Excitation-Emission Maps

Excitation-Emission Maps (EEMs) provide a ‘Finger Print’ of complex mixtures of substances. These maps are typically measured either by a series of emission scans with stepwise increase, or for synchronous maps, by a series of synchronous scans and stepwise increase of the excitation-emission offset. A map measurement over a wide range of excitation and emission wavelengths, as shown here, can only be performed properly if higher order scatter is automatically removed during the measurement; the FS5’s built in automatic filters, along with real-time correction for background noise and spectral efficiencies, allow these measurements to be seamlessly performed by even the newest users.

EEM

Measurements of absolute fluorescence quantum yield

Fluorescence quantum yields can be measured by using the optional, sample chamber mounted, integrating sphere. The absolute method directly compares the number of absorbed and emitted photons through measurement of a blank-reference and a sample where their spectra can be integrated with respect to each other. The quantum yield calculation is made using a wizard within the operating software.

QY

Quinine bisulphate in Perchloric acid. The red curve shows the scan over the excitation scatter at 350 nm and the emission of the sample, the blue curve shows the scatter of the blank measurement. The sample and blank-reference emission range (370 – 700 nm) has been increased by a factor 100 for better demonstration.

Chromaticity and Colour Co-ordinates

The lighting industry requires precise determination of the colour co-ordinates of fluorescent powders. The FS5 and Fluoracle Software provides Chromaticity analysis tools for the determination of colour co-ordinates and luminosity values using CIE 1931 and CIE 1976. The example shows four commercial powders with blue, green, yellow and red emission.

CIE 1931, CIE 1976CIE 1931, CIE 1976CIE 1931, CIE 1976

Plate Reader

Multiple sample measurements can be made using the plate reader sample holder accessory. This can be used for liquid samples with commercial plates of up to 96 wells, but is also suitable for routine quality assessment of fluorescent powders. As with all the sample holder accessories, this is also compatible with the FS5 upgrade options, including the lifetime upgrades.

Micro-wellMicro-well

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