Bioresource Technology, Vol.161, 149-154, 2014
Study of cellular development and intracellular lipid bodies accumulation in the thraustochytrid Aurantiochytrium sp KRS101
Transmission electron, confocal microscopy and FACS in conjunction with two different lipophilic fluorescent dyes, BODIPY 505/515 and Nile Red were used to describe the cellular development and lipid bodies formation in Aurantiochytrium sp. KRS101. TEM results revealed that multi-cellular spores were appeared in sporangium during early-exponential phase, and spores were matured in mid-exponential phase followed by release of spores from sporangium in late-exponential phase. TEM and FACS analyses proved that lipid bodies appeared, developed and degenerated in mid-exponential, early- and late-stationary phases, respectively. The staining results in FACS indicate that BODIPY 505/515 is more effective for the vital staining of intracellular lipid bodies than Nile Red. FACS based single cell sorting also showed healthy growth for BODIPY 505/515 stained cells than Nile Red stained cells. In addition, a quantitative baseline was established either for cell growth and/or lipid accumulation based on cell count, fatty acid contents/composition, and sectional/confocal images of KRS101. (C) 2014 Elsevier Ltd. All rights reserved.
Keywords:Aurantiochytrium sp.;TEM analysis;BODIPY 505/515;Nile Red;Fluorescence-activated cell sorting