ABSTRACT
This chapter presents the basics of transmission/absorption measurements that are qualified as steady-state experiments. In most cases, the absorption of a specific chromophore is measured in a well-defined buffer and under well-defined conditions. A single absorption/transmission measurement requires two measurements: a reference measurement and a sample measurement. Transmission and transmittance are used alternatively in describing the transmission of the light. The transmittance of light depends on the type of transparent material being used, as different materials will have different wavelength ranges through which light will efficiently permeate. The biggest factor contributing to the attenuation of light by clear transparent materials is a reflection on the interface. It’s important to realize that the extent of light attenuation by transparent materials mostly depends on the type of material. The shape of the absorption spectrum should be independent of the sample concentration or from the cuvette used for measurement.
