Fluorometer

A simplistic design of the components of a fluorometer

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.

Light sources for fluorometers are often dependent on the type of sample being tested. Among the most common light source for fluorometers is the low-pressure mercury lamp. This provides many excitation wavelengths, making it the most versatile. However, this lamp is not a continuous source of radiation. The xenon arc lamp is used when a continuous source of radiation is needed. Both of these sources provide a suitable spectrum of ultraviolet light that induces chemiluminescence. These are just two of the many possible light sources.

Glass and silica cuvettes are often the vessels in which the sample is placed. Care must be taken to not leave fingerprints or any other sort of mark on the outside of the cuvette, because this can produce unwanted fluorescence. "Spectro grade" solvents such as methanol are sometimes used to clean the vessel surfaces to minimize these problems.

There are two basic types of fluorometers, the filter fluorometer and the spectrofluorometer. The difference between them is the way they select the wavelengths of incident light. A filter fluorometer will use filters while a spectrofluorometer will use grating monochromators. Filter fluorometers are often purchased/built at a lower cost but are less sensitive and have less resolution than spectrofluorometers. Also, a filter fluorometer is, obviously, only capable of operation at the wavelenth(s) of the available filter(s), whereas a monochromator is generally freely tunable over a relatively wide range. The potential disadvantege of a monochromator arises from that same property, because the monochromator is capable of miscalibration or misadjustment, where the wavelength of a filter is fixed when it is manufactured.

• Filter fluorometer
• Spectrofluorometer
• Integrated fluorometer

• Fluorescence spectroscopy, for a fuller discussion of instrumentation
• Chlorophyll fluorescence, to investigate plant ecophysiology.
• Integrated fluorometer to measure gas exchange and chlorophyll fluorescence of leaves.
• Radiometer, to measure various electromagnetic radiation
• Spectrometer, to analyze spectrum of electromagnetic radiation
• Scatterometer, to measure scattered radiation
• Microfluorimetry, to measure fluorescence on a microscopic level
• Interference filter, thin film filters that work by optical interference, showing how they can be tuned in some cases