Measuring photosynthetic parameters in individual algal cells by Fast Repetition Rate fluorometry

A Single Cell Fast Repetition Rate (SCFRR) fluorometer was developed to measure the quantum yield of photochemistry, the functional absorption cross section of PS II and the kinetics of electron transport on the acceptor side of PS II in individual algal cells. These parameters are used to quantify the cell-specific photosynthetic performance in natural phytoplankton assembledges in aquatic ecosystems. The SCFRR technique measures chlorophyll fluorescence transients induced by a precisely controlled series of excitation flashlets that cumulatively saturate PS II within 120 mu s. To meet the requirement in the analysis for single algal cells, the measurements are conducted in micro volumes, such that the probability of probing more than one cell at a time is vanishingly low. We designed a novel, computer-controlled hydromechanical system to deliver a portion of the sample into the measuring chamber and, following measurement, remove it into one of six sorting containers. The fluorescence signal is induced by a series of high frequency flashlets obtained from high luminosity blue light-emitting diodes and is acquired by a novel red-sensitive PMT-based detection system exhibiting both high sensitivity and a very wide dynamic range. The wide dynamic range of the detector allows SCFRR measurements for a wide variety of cell sizes ranging from 1 to 100 mu m equivalent spherical diameter. The compact and light-weight design makes the SCFRR Fluorometer applicable for both laboratory and field studies.