fluorometría

Fluorometría is a sensitive analytical technique used to measure the fluorescence emitted by substances when exposed to specific wavelengths of light. The method relies on the principle that certain molecules, known as fluorophores, absorb light at one wavelength and re-emit it at a longer wavelength, a process called Stokes shift. This phenomenon allows fluorometry to detect and quantify even trace amounts of substances with high precision.

The basic components of a fluorometer include a light source (often a lamp or laser), a monochromator to select excitation wavelengths, a sample holder, and a detector to measure emitted light. Samples may be in liquid, solid, or gaseous form, depending on the application. Common fluorophores include fluorescent dyes, proteins like GFP (Green Fluorescent Protein), and naturally occurring molecules such as chlorophyll or vitamins.

Fluorometry finds applications in diverse fields, including biochemistry, environmental science, and clinical diagnostics. In biochemistry, it is used to study protein interactions, nucleic acid hybridization, and enzyme kinetics. Environmental monitoring employs fluorometry to detect pollutants or measure organic matter in water samples. Clinically, it aids in diagnosing diseases through the analysis of biomarkers or cellular components.

The technique offers advantages such as high sensitivity, selectivity, and the ability to perform non-destructive measurements. However, factors like quenching (reduction of fluorescence intensity) and background interference can affect accuracy, requiring careful experimental design and calibration. Modern fluorometers often incorporate advanced features like time-resolved fluorescence and imaging capabilities to enhance analytical capabilities.