10/13/09 1 Lecture 14 – Oct 14 Chem 4101 – Fall 2009 Fluorescence 1- Why is fluorescence more sensitive than UV-VIS absorption? 2- Photoluminescence methods 3- Chemiluminescence -Sections 15C-15D Suggested exercises: 15-15, 15-12, 15-9, obtain fluorescence spectrum of fluorescein Lecture 14 – Oct 14 Chem 4101 – Fall 2009 Fluorescence versus UV-VIS absorption Photon noise For n photons counted in a given time The standard deviation (noise) is n1/2 S/N = n/(n1/2) = n1/2 Absorption no photons/s (blank) n photons/s (sample) Signal = no – n Noise ≈ (2no)1/2 S/N ≈ (1-T) (no/2)1/2 Fluorescence nf = 2.303φK’’εbcno fluorescence photons Noise = (nf)1/2 S/N = (2.303φK’’εbcno) 1/2 Lecture 14 – Oct 14 Chem 4101 – Fall 2009 Determination of inorganic species Figure 15-15, Section 15C Table 15-210/13/09 2 Lecture 14 – Oct 14 Chem 4101 – Fall 2009 Derivatization of functional groups www.invitrogen.com www.invitrogen.com/site/us/en/home/References/Molecular-Probes-The-Handbook.html Alexa Fluor® 350 carboxylic acid, succinimidyl ester (fluorescent reagent) 3-(2-furoyl)quinoline-2-carboxaldehyde (fluorogenic reagent) Lecture 14 – Oct 14 Chem 4101 – Fall 2009 Chemiluminescence Figure 15-17, Section
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