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.
Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Natarajan
N
Velmurugan
Department of Forest Science and Technology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonju, Republic of Korea.
Yesupatham
Y
Sathishkumar
Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Sung Sun
SS
Yim
Department of Forest Science and Technology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonju, Republic of Korea.
Yang Soo
YS
Lee
Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea; Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, USA.
Min S
MS
Park
Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Ji Won
JW
Yang
Department of Chemical and Biomolecular Engineering (BK21 Program), KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea; Institute for the BioCentury, KAIST, Daejeon, Republic of Korea. Electronic address: kjjeong@kaist.ac.kr.
Ki Jun
KJ
Jeong
D001896:Boron Compounds; D005227:Fatty Acids / Q000032:analysis; D005410:Flame Ionization; D005434:Flow Cytometry; D066292:Lipid Droplets / Q000502:physiology*; D050356:Lipid Metabolism; D010078:Oxazines; D058009:Stramenopiles / Q000737:chemistry* / Q000502:physiology* / Q000648:ultrastructure
C000594973:BODIPY 505-515; D001896:Boron Compounds; D005227:Fatty Acids; D010078:Oxazines; C044808:nile red
Aurantiochytrium sp.; BODIPY 505/515; Fluorescence-activated cell sorting; Nile Red; TEM analysis