title: Synchrotron-FTIR microspectroscopy enables the distinction of lipid accumulation in thraustochytrid strains through analysis of individual live cells.
abstract: The superior characteristics of high photon flux and diffraction-limited spatial resolution achieved by synchrotron-FTIR microspectroscopy allowed molecular characterization of individual live thraustochytrids. Principal component analysis revealed distinct separation of the single live cell spectra into their corresponding strains, comprised of new Australasian thraustochytrids (AMCQS5-5 and S7) and standard cultures (AH-2 and S31). Unsupervised hierarchical cluster analysis (UHCA) indicated close similarities between S7 and AH-7 strains, with AMCQS5-5 being distinctly different. UHCA correlation conformed well to the fatty acid profiles, indicating the type of fatty acids as a critical factor in chemotaxonomic discrimination of these thraustochytrids and also revealing the distinctively high polyunsaturated fatty acid content as key identity of AMCQS5-5. Partial least squares discriminant analysis using cross-validation approach between two replicate datasets was demonstrated to be a powerful classification method leading to models of high robustness and 100% predictive accuracy for strain identification. The results emphasized the exceptional S-FTIR capability to perform real-time in vivo measurement of single live cells directly within their original medium, providing unique information on cell variability among the population of each isolate and evidence of spontaneous lipid peroxidation that could lead to deeper understanding of lipid production and oxidation in thraustochytrids for single-cell oil development.
authors.affiliation: Centre for Chemistry and Biotechnology (CCB), School of Life and Environmental Sciences, Deakin University, Pigdons Road, Waurn Ponds, Victoria 3216, Australia. Electronic address: p.vongsvivut@deakin.edu.au.
authors.forename: Jitraporn
authors.initials: J
authors.lastname: Vongsvivut
authors.affiliation: Centre for Biospectroscopy, School of Chemistry, Monash University, Wellington Road, Clayton, Victoria 3800, Australia.
authors.forename: Philip
authors.initials: P
authors.lastname: Heraud
authors.affiliation: Centre for Chemistry and Biotechnology (CCB), School of Life and Environmental Sciences, Deakin University, Pigdons Road, Waurn Ponds, Victoria 3216, Australia.
authors.forename: Adarsha
authors.initials: A
authors.lastname: Gupta
authors.affiliation: Centre for Chemistry and Biotechnology (CCB), School of Life and Environmental Sciences, Deakin University, Pigdons Road, Waurn Ponds, Victoria 3216, Australia.
authors.forename: Tamilselvi
authors.initials: T
authors.lastname: Thyagarajan
authors.affiliation: Centre for Chemistry and Biotechnology (CCB), School of Life and Environmental Sciences, Deakin University, Pigdons Road, Waurn Ponds, Victoria 3216, Australia.
authors.forename: Munish
authors.initials: M
authors.lastname: Puri
authors.affiliation: Centre for Biospectroscopy, School of Chemistry, Monash University, Wellington Road, Clayton, Victoria 3800, Australia.
authors.forename: Don
authors.initials: D
authors.lastname: McNaughton
authors.affiliation: Centre for Chemistry and Biotechnology (CCB), School of Life and Environmental Sciences, Deakin University, Pigdons Road, Waurn Ponds, Victoria 3216, Australia. Electronic address: cbarrow@deakin.edu.au.
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Barrow
mesh_terms: D056808:Biostatistics; D005227:Fatty Acids / Q000032:analysis*; D059010:Single-Cell Analysis / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D058009:Stramenopiles / Q000737:chemistry* / Q000145:classification*
chemical_list: D005227:Fatty Acids
keywords: Thraustochytrids; lipids; multivariate data analysis.; polyunsaturated fatty acids; synchrotron Fourier transform infrared (S-FTIR) microspectroscopy