Explain the theory of fluorescence spectroscopy and its applications in biology.

Basic Clinical Laboratory Techniques 6E
6th Edition
ISBN:9781133893943
Author:ESTRIDGE
Publisher:ESTRIDGE
Chapter7: Basic Clinical Microbiology
Section7.7: Bacterial Identification And Antibiotic Susceptibility Testing
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Explain the theory of fluorescence spectroscopy and its applications in biology.

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Step 1 Introduction:

Fluorescence is the molecular absorption of light energy at one wavelength and its nearly instantaneous re-emission at another, usually longer, wavelength.

Step 2 Theory:

Fluorescence is a type of luminescence caused by photons exciting a molecule, raising it to an electronically excited state. It's brought about by absorption of photons in the singlet ground state promoted to a singlet-excited state.

If the incident light is at a wavelength where the molecule will absorb the photon, the molecule is then excited from the electronic ground state to a higher excited state.

The electrons then go through internal conversion, affected by vibrational relaxation and heat loss to the environment. It then emits a photon from the lowest-lying singlet excited state in the form of fluorescence.

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