Fig. 1

The workflow employed in lipid characterization of microalgae. Bioprospecting of aquatic and soil algae, as well as mutagenesis of algal cells, such as C. reinhardtii, are carried out to generate algal samples with potentially desirable lipid characteristics. Mutagenized cells are sorted by FACS, based on fluorescence of a lipophilic dye to isolate cells with increased lipid accumulation phenotypes. The obtained environmental samples and screened mutants are then analyzed by confocal Raman microscopy. This method, once optimized, allows for rapid in situ characterization of lipids through ratiometric analysis of Raman spectra. As a rapid process, this workflow offers better spatial resolution and characterization of about 10 cells per hour. The spectra yield information about the number of C=C bonds and the hydrocarbon chain length of the lipid molecules. The workflow allows rapid characterization of algae for molecular traits that are suitable for use in production of biofuels