Fluorescence and phosphorescence

Possible de-excitation pathways of excited molecules.

Various parameters influence the emission

of fluorescence.
 Fluorescent probes

Compounds whose fluorescence doesn´t change after their interaction with

biological material:
acridine orange (DNA)
fluorescein (proteins)
rhodamine (proteins)
...

Compounds whose fluorescence change according to their environment:

ANS (1-anilinonaftalen-8- sulphonate) - polarity

Fura-2 - tracking the movement of calcium within cells
...

The larger the fluorescence quantum yield, the easier it is to observe a

fluorescent compound, especially a fluorescent probe.
 Some applications of fluorescence detection

• Protein conformation
• Membrane potential
• Membrane transport
• Membrane viscosity
• Enzymatic reactions
• DNA analysis
• Genetic engineering (manipulations)
• Immunochemical methods
• Cell proliferation and apoptosis

Fluorescence and phosphorescence are one

of the most sensitive techniques available!