FLUORESCENCE

Analytics Making Sense of this Technology

Electromagnetic Waves
The electromagnetic spectrum describes all possible light, which is characterized by wavelength and frequency. A longer wavelength () means a lower frequency (v) and lower energy (E). Photons transmit the energy of the electromagnetic wave.

Equations

c: speed of light in a vacuum, 2.998*108 m/s h: Planks constant, 6.626*10-34 Js

Florescence is a type of electromagnetic radiation that involves the emission of photons to produce visible light. Molecules that undergo this are called fluorophores.

Fluorescence Process

2) The electron remains in this excited state S1 for a few

nanoseconds as some energy is released to S1.

1) First, a photon of high

energy is absorbed by the fluorophore, exciting its electron from ground state S0 to a higher energy state.

3) The molecule emits a

photon of lower energy, causing it to return to the ground state S0.

Jablonski diagram of the electronic states in the fluorescence process

4) This process is repeated until the fluoresce is lost, an effect called photobleaching.

Excitation Light Source

Excitation Monocrometer: Takes light at many wavelengths and filters to one wavelength

Sample Cell: Emits photons

Detection Device
The detector must be at a right angle from the light source so that the excitation light does affect the reading. The detector also must be calibrated with standards before use.

Emission Monocrometer

Fluorescence Detector: Records emission photons

Industry
Corrosive INHIBITOR

Applications Research
Drug Delivery
The fluorescent tag dims when an inhibitor is attached to a corrosive material.

Corrosion Inhibition

Corrosive

Fluorescently Labeled Drug Molecule

Drug Delivery System

InStream Data Saves Money Uses Less Inhibitor Better Control

1
Fluorescently labeled drug diffuses into solution over time

2
Concentration of drug in solution can be measured

Drug released over time can be determined through concentrations of fluorescent molecules (Release Rate)

Wastewater Analysis

DNA Detection

Used to measure the amount of oil and grease in waste water for petroleum processing
Fluoresencesce Fundamentals. Life Technologies. [Online].

Molecules from dyes such as Eva Green and Sybr Green bind to DNA and emit florescent light. This is used in Real Time Polymerase Chain Reaction (PCR) to quantify the amount of DNA that has been replicated.
Malkov, Vladmir, and Dietmar Sievert. "Oil-in-Water Fluorescence Sensor in Wastewater and Other Industrial Applications." PPChem (n.d.): n. pag. Web. Corrosion Inhibition." Cooling Water Application NALCO Best Practices. Vol. 313. N.p.: n.p., n.d. 365-412. Print.

http://www.lifetechnologies.com/us/en/home/references/molecular-probes-thehandbook/introduction-to-fluorescence-techniques.html D. Harris, Fundamentals of Spectrophotometry, in Quantitative Chemical Analysis. New York: W.H. Freeman, 2010.