title: Application of high-content image analysis for quantitatively estimating lipid accumulation in oleaginous yeasts with potential for use in biodiesel production.
abstract: Biodiesel from oleaginous microorganisms is a viable substitute for a fossil fuel. Current methods for microorganism lipid productivity evaluation do not analyze lipid dynamics in single cells. Here, we described a high-content image analysis (HCA) as a promising strategy for screening oleaginous microorganisms for biodiesel production, while generating single-cell lipid dynamics data in large cell density. Rhodotorula slooffiae yeast were grown in standard (CTL) or lipid trigger medium (LTM), and lipid droplet (LD) accumulation was analyzed in deconvolved confocal microscopy images of cells stained with the lipophilic fluorescent Nile red (NR) dye using automated cell and LD segmentation. The 'vesicle segmentation' method yielded valid morphometric results for limited lipid accumulation in smaller LDs (CTL samples) and for high lipid accumulation in larger LDs (LTM samples), and detected LD localization changes. Thus, HCA can be used to analyze the lipid accumulation patterns likely to be encountered in screens for biodiesel production.
authors.affiliation: Laboratory of Biotechnology - Labio, Directory of Metrology Applied to Life Science - Dimav, National Institute of Metrology, Quality and Technology - Inmetro, Duque de Caxias, RJ, Brazil; Agrocampus Ouest, Rennes, France; Université Rennes, Rennes, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Capus
authors.affiliation: Laboratory of Biotechnology - Labio, Directory of Metrology Applied to Life Science - Dimav, National Institute of Metrology, Quality and Technology - Inmetro, Duque de Caxias, RJ, Brazil.
authors.forename: Marianne
authors.initials: M
authors.lastname: Monnerat
authors.affiliation: Laboratory of Biotechnology - Labio, Directory of Metrology Applied to Life Science - Dimav, National Institute of Metrology, Quality and Technology - Inmetro, Duque de Caxias, RJ, Brazil.
authors.forename: Luiz Carlos
authors.initials: LC
authors.lastname: Ribeiro
authors.affiliation: Laboratory of Cellular Ultrastructure Hertha Meyer, Federal University of Rio de Janeiro, Rio de Janeiro, RJ, Brazil; National Institute of Structure Biology and Bioimaging, Rio de Janeiro, RJ, Brazil.
authors.forename: Wanderley
authors.initials: W
authors.lastname: de Souza
authors.affiliation: Laboratory of Biotechnology - Labio, Directory of Metrology Applied to Life Science - Dimav, National Institute of Metrology, Quality and Technology - Inmetro, Duque de Caxias, RJ, Brazil.
authors.forename: Juliana Lopes
authors.initials: JL
authors.lastname: Martins
authors.affiliation: Laboratory of Biotechnology - Labio, Directory of Metrology Applied to Life Science - Dimav, National Institute of Metrology, Quality and Technology - Inmetro, Duque de Caxias, RJ, Brazil; National Institute of Structure Biology and Bioimaging, Rio de Janeiro, RJ, Brazil. Electronic address: cbfilho@inmetro.gov.br.
authors.forename: Celso
authors.initials: C
authors.lastname: Sant'Anna
mesh_terms: D000465:Algorithms; D056804:Biofuels; D003246:Conservation of Energy Resources; D007091:Image Processing, Computer-Assisted / Q000379:methods; D066292:Lipid Droplets / Q000648:ultrastructure; D008055:Lipids / Q000096:biosynthesis*; D012248:Rhodotorula / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D056804:Biofuels; D008055:Lipids
keywords: Biodiesel; Biofuel; High-content image analysis; Lipid droplets; Oleaginous microorganisms