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Western University
Biology 2290F/G

DeanUnit Diagram the Spectronic 20 optical system Explain how the instrument works o Tungsten lamp provides radiant energy of wavelength 340 to 900 nm o Light is focused on a diffraction grating which splits the light into its component wavelengthsChanging the position of the diffraction grating allows you to choose light of a very narrow range of wavelength to pass through the slit before reaching the sample o Any light that is transmitted not absorbed reaches a phototube that converts the light energy into an electric current o Measures transmittance of a given sample and compares it to a reference blank to give an absorbance valueDescribe transmittance and absorbanceo Transmittancethe fraction of incident light that passes through the sample o Absorbanceamount of light absorbed by the sampleConvert transmittance values to absorbance values o Alog1TState Lamberts Law and Beers Law in words o Lamberts lawfor a thin layer sample the same proportion of light is absorbed regardless of the power of the incident light o Beers lawabsorbance of a solution is proportional to the number of absorbing particles or concentration of absorbing material in solutionA EclIdentify an appropriate blank to zero the spectrophotometer for a given experiment o Everything contained in the experimental solution minus the compound of interestGive examples of the principal uses of the spectrophotometer lab notes p 13 o Absorption spectra o Standard curve o Rate of a biochemical reactionDescribe how to prepare absorption spectra and how they are used o Absorbance values yaxis of a solution plotted against various wavelengths xaxisDescribe how to prepare and use a standard curve of Absorbance vs concentration Interpret A values o Standard curvedone at max wavelength and used to find E cA o Achange in absorptionUse A values to calculate reaction ratesExplain the meaning of resolution in microscopy and the theoretical limit to resolution by reference to Abbes equation Resource Manual pp 4749 o Resolution is the smallest distance by which two objects can be separated in order to allow them to be imaged as two separate objects
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