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Fluorescence and Phosphorescence: Instrumentation

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.

Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.

Photoluminescence: Applications

Photoluminescence: Applications

Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...

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Related Experiment Video

Updated: Jun 16, 2026

Fluorescence detection methods for microfluidic droplet platforms

Fluorescence detection methods for microfluidic droplet platforms

Published on: December 10, 2011

High sensitivity fluorophotometer using photon counting.

R Aoshima, K Iriyama, H Asai

|February 4, 2010

Summary

This summary is machine-generated.

A new photon-counting apparatus detects very weak fluorescence, showing linear sensitivity for riboflavin solutions. This sensitive instrument also measured emission spectra for chlorophyll monolayers with high signal quality.

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Area of Science:

Analytical Chemistry
Biophysics
Spectroscopy

Background:

Detecting weak fluorescence is crucial for analyzing low-concentration substances.
Traditional methods may lack sensitivity for trace-level molecular detection.
Photon counting offers a potential pathway to enhanced fluorescence detection.

Purpose of the Study:

To develop and validate a photon-counting apparatus for sensitive fluorescence detection.
To assess the apparatus's performance using riboflavin as a standard.
To measure the emission spectra of chlorophyll monolayers.

Main Methods:

Construction of a photon-counting based fluorescence detection apparatus.
Measurement of fluorescence intensity from aqueous riboflavin solutions across a range of concentrations.
Acquisition of emission spectra from chlorophyll a and b monolayers.

Main Results:

The apparatus demonstrated linear detection of riboflavin fluorescence from 4 x 10(-11) g/ml to 1 x 10(-6) g/ml.
High signal-to-noise ratios were achieved for chlorophyll a and b emission spectra.
The system proved effective for detecting very weak fluorescence signals.

Conclusions:

The developed photon-counting apparatus is highly sensitive and suitable for quantifying trace amounts of fluorescent compounds.
The instrument provides reliable spectral data for biological pigments like chlorophyll.
This technology advances capabilities in sensitive fluorescence spectroscopy.