Fluorescence Spectroscopy

fluorescence spectroscopy

Fluorescence spectroscopy, also known as fluorometry or spectrofluorometry, is a type of electromagnetic spectroscopy, which analyses fluorescence from a sample.

 

It involves using a beam of light tp excite the electrons in molecules of certain compounds and causes them to emit light which is typically, but not necessarily, visible light. It is a useful technique in many biological (chlorophyll and carotenoid), biochemical (fluorescence diagnosis of malignancies) and environmental applications.

 

For most fluorescence applications, the amount of fluorescence energy emitted is only around 3% of the amount of excitation light energy. Fluorescence light also has a lower energy (higher wavelength) than the excitation energy and is usually scattered light - this means it emits energy in all directions.

For optimal performance, assuming the time acquisition window is not limited, we recommend our AvaSpec-ULS2048LTEC spectrometer for this application. It can support long integration times often exceeding 5 seconds. When higher-speed acquisition is required, we recommend the AvaSpec-ULS2048XL back-thinned CCD spectrometer. For maximum sensitivity, the SensLine range spectrometerer AvaSpec-HS1024x122-USB2 is recommended.

When configuring the measurement setup, preventing excitation light from entering the spectrometer is an crucial issue. Possible methods to accomplish this, where one does not exclude the other, include:

  • Make use of an AvaLight-LED light source which typically has a narrow bandwidth enabling the limitation of excitation to shorter wavelengths that are not part of the emission spectrum

  • Use a broadband light source such as the AvaLight-HAL-Mini for high output in combination with an (interference) bandpass or lowpass filter.

  • Make sure the optical path for excitation light and fluorescence are perpendicular - this means the excitation light will not enter the receiving fibre (use the CUV-UV/VIS-FL or the CUV-DA)

  • Use the fluorescence decay time to separate excitation energy from the integration time start pulse. A pulsed light source can be used for this (pulsed laser or AvaLight-XE Xenon flash)

fluorescence spectroscopy

Fluorescence setup with enhanced Probe

fluorescence spectroscopy

Fluorescence setup with Micro Flow Cell

fluorescence spectroscopy

Fluorescence cuvette setup

with blocking filter

fluorescence spectroscopy

Fluorescence cuvette setup

without blocking filter

Ava-Fluorescence System
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