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Introduction to Spectroscopy and Fluorescence

Introduction to Spectroscopy and FluorescenceFor students of HI 6001-125 Computational Structural Biology Willy Wriggers, from material by Mathew Baker, Univ. of E UN I V E R S I T Y of TE X A SSC H O O L O F HE A L T H IN F O R M A T I O NSC I E N C E S A T HO U S T O NWhat is Spectroscopy ? By using different parts of the electromagnetic spectrum (different types of light) we can see separate details. The size and energy of the light waves restrict their uses, as does our ability to measure & interpret them. Redefine how you think of seeing . WavelengthFrequency EnergyLoss of lightProduction of lightIntensity of diffracted beamAbsorptionEmissionScattering/Diffrac tion SourceSampleDetectorInteractions with SampleA ZXYA =Amplitude = frequency = WavelengthPhaseElectro-Magnetic Radiation The Frequency ( ) & wavelength ( ) of a wave are related by: = c/ (c= 3 x 108 ) Frequency can be converted to energy (e) by: e = h (h= x ) Scales: 1 nm = 10-7cm = 10 Electro-Magnetic RadiationEnergy = hc The circles represent energy levels - representing increasing distances from the nucleus.

Introduction to Spectroscopy and Fluorescence ... monitoring Trp fluorescence is the same as the pK for a known ionisable group (e.g. a carboxyl) then the group must be near the Trp. •If a substance that effects the Q of the free amino acid doesn’t effect the fluorescence of the

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