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Concept
Fluorometer
Summary
A fluorometer, fluorimeter or fluormeter is a device used to measure parameters of visible spectrum fluorescence: its intensity and wavelength distribution of emission spectrum after excitation by a certain spectrum of light. These parameters are used to identify the presence and the amount of specific molecules in a medium. Modern fluorometers are capable of detecting fluorescent molecule concentrations as low as 1 part per trillion.
Fluorescence analysis can be orders of magnitude more sensitive than other techniques. Applications include chemistry/biochemistry, medicine, environmental monitoring. For instance, they are used to measure chlorophyll fluorescence to investigate plant physiology.
Typically fluorometers utilize a double beam. These two beams work in tandem to decrease the noise created from radiant power fluctuations. The upper beam is passed through a filter or monochromator and passes through the sample. The lower beam is passed through an attenuator and adjusted to try and match the fluorescent power given off from the sample. Light from the fluorescence of the sample and the lower, attenuated beam are detected by separate transducers and converted to an electrical signal that is interpreted by a computer system.
Within the machine the transducer that detects fluorescence created from the upper beam is located a distance away from the sample and at a 90-degree angle from the incident, upper beam. The machine is constructed like this to decrease the stray light from the upper beam that may strike the detector. The optimal angle is 90 degrees.
There are two different approaches to handling the selection of incident light that gives way to different types fluorometers. If filters are used to select wavelengths of light, the machine is called a fluorometer. While a spectrofluorometer will typically use two monochromators, some spectrofluorometers may use one filter and one monochromator. Where, in this case, the broad band filter acts to reduce stray light, including from unwanted diffraction orders of the diffraction grating in the monochromator.
Official source
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