Versatile spectrophotometer for photosynthesis (light-induced changes in absorbance and fluorescence yield, circular and linear dichroism) and other biophysical measurements

Abstract

The construction of a versatile, single-beam spectrophotometer expanded from a previously reported circular dichrometer utilizing an elasto-optic modulator (EOM) for producing circular polarization, is described. The new spectrophotometer is capable of measuring circular dichroism as well as linear dichroism and light-induced changes in absorbance and fluorescence yield, the latter being two of the most commonly measured properties in photosynthesis research. The use of an EOM, along with a phase-sensitive detector, affords high sensitivity and at the same time, in conjuction with a mechanical phosphoroscopic device, a high degree of immunity from artifacts in studies employing actinic light. A description is given of the generation and use of timing signals essential for control and signal processing. Its performance is illustrated by examples drawn from light-induced absorbance and fluorescence-yield changes associated with photosynthetic electron transport and changes in redox state of electron carriers. This photometer, with very little modification, may also be used to perform a host of other types of measurements.