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Fluorescence Spectroscopy

Many organic compounds fluoresce, which is to say they emit light (usually visible but not always) when their electrons are excited by electromagnetic radiation of a lower wavelength. The amount of fluorescence energy emitted is only a small fraction of the energy input, typically only about 3% of excitation, and is more diffuse, scattering light in all directions. These conditions require a powerful spectrometer with high-sensitivity and long integration times to capture fluorescence data.

Fluorescence Spectroscopy, also called fluorometry or spectrofluorometry, is a very powerful tool. However, one important concern when applying this measurement technique is the possibility for stray excitation light to enter the spectrometer. Avantes offers several ways to mitigate this stray excitation light. One might employ a narrow bandwidth AvaLight-LED source to limit excitation wavelengths to those not part of the emission spectrum, or choose an AvaLight-Hal high-output broadband light source along with a bandpass or low-pass filter. A more mechanical solution might be to ensure the light can’t enter the fiber leading into the spectrometer with the use of a cuvette holder (use the CUV-UV/VIS-FL or the CUV-DA) to ensure the excitation path is perpendicular to the optical path for fluorescence measurements. Finally, it is possible to use the rate of fluorescence decay to separate the excitation-emission from the fluorescence measurement using a pulsed light source such as the AvaLight-XE Xenon flash and triggering options on board the spectrometer.

Avantes offers a basic fluorescence bundle that includes an AvaSpec-ULS2048L high-sensitivity spectrometer along with LED light source and fluorescence probe. For applications that need even higher sensitivity, Avantes offers several options including the AvaSpec-Hero (HSC1024x58Tec-EVO) which perfectly balances high-sensitivity and high-resolution in this TE cooled CCD back-thinned spectrometer.

Avantes also offers a wide selection of fiber optic sampling accessories for fluorescence spectroscopy applications including the cuvette holders, direct attached and in-line filter holders and flow cells to support any fluorescence application need.

Applications for Fluorescence Measurements

There are numerous uses for fluorescence spectroscopy, but one of the largest areas is in the life sciences and biomedical field. Fluorescence spectroscopy is used to diagnose malignancies because cancer cells do not have the same fluorescence response that healthy tissue does, but it is also used to tag genetic markers to facilitate genetic research.

Fluorescence can also be utilized in agricultural applications due to the fluorescent response of chlorophyll and fluorescent dyes can be employed for security measures such as identifying leaking radioactive byproducts in the generation of nuclear electric power.

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Fluorescence Instruments

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AvaSpec-ULS2048CL-EVO

The StarLine AvaSpec-ULS2048CL-EVO offers you this latest technology using CMOS instead of the conventional CCD technology. In combination with our latest AS-7010 electronics, it offers you a versatile device including USB3.0 communication with 10x higher speed communications and 50x more memory. The AvaSpec-ULS2048CL-EVO is also available as OEM unit, bench-only or rackmount version.

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AvaSpec-HSC2048XL-EVO

The AvaSpec-HS2048XL-EVO is an exceptional instrument for high sensitivity applications where high resolution is not of paramount concern. It features the Avantes HS optical bench which has a full 0.22 numerical aperture for superior throughput The AvaSpec-HS2048XL-EVO uses the AS7010 electronics board offering USB3 (10 times faster than USB2), Gigabit Ethernet and better signal processing.

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AvaSpec-HS1024x58-TEC (AvaSpec-Hero)

This spectrometer combines the best of both worlds: high resolution and high sensitivity! The AvaSpec-HERO is build up around our High Sensitivity Compact 100mm optical bench, offering a NA of 0.13 and a cooled Backthinned Detector (1024x58 pixels). The Hero uses the newest AS7010 board, including a High Performance Analog to Digital convertor

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Fluorescence Resources

Fluorescence Information Page

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Download: Fluorescence Application Note

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Fluorescent Measurement Instrument Bundle

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Video: How to do a Fluorescence Measurement

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Fluorescence System Design with Beam Splitter/Combiner

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Fluorescence by Avantes

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