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Industrial Microbiology 2016
October 17-18, 2016
Volume 8, Issue 5(Suppl) J Microb Biochem Technol
ISSN:1948-5948 JMBT, an open access journal
conferenceseries .com
October 17-18, 2016 Kuala Lumpur, Malaysia
Industrial & Pharmaceutical MicrobiologyInternational Conference and Summit on
Hemanth Nag Noothalapati Venkata, J Microb Biochem Technol 2016, 8:5(Suppl)
Stable isotope probing coupled Raman microscopy:An efficient way to study single cell biochemistry
Hemanth Nag Noothalapati Venkata
Shimane University, Japan
Lipid droplets have been hypothesized to be intimately associated with intracellular proteins. However, there is little direct
evidence for both spatiotemporal and functional relations between lipid droplets and proteins provided by molecular -level
studies on intact cells. To elucidate the interplay between them at the single cell level, Raman microscopy was coupled with a
very powerful strategy, namely, stable isotope labeling. Here, I present in vivo time lapse Raman imaging, coupled with stable-
isotope (13C) labeling, of single living Schizosaccharomyces pombe cells. Our results show that the proteins newly synthesized
from incorporated 13C-substrate are localized specifically to lipid droplets as the lipid concentration within the cell increases.
Lipids, which help to store energy in a compact form, have variety of roles in biological systems and their metabolism is central
to life. Here, we show that combination of stable isotope probing (SIP), Raman micro-spectroscopy and multivariate curve
resolution analysis can serve as a valuable approach in metabolomics research. We studied ergosterol biosynthesis in single
living fission yeast cells, grown in mixtures of normal (12C) and 13C-glucose as the sole carbon source. By carefully looking
into the biosynthetic pathways and by comparing the observed peak positions with calculation results on isotope-substituted
ergosterol, it is possible to understand how 13 C is incorporated in the conjugated C=C moiety of the molecule. The multivariate
spectral data analysis revealed intrinsic spectra and their relative abundances of all isotopomers.
Biography
Hemanth Nag Noothalapati Venkata has completed his PhD from National Chiao Tung University, Taiwan. During his PhD, he studied spatio-temporal relationship
between proteome and lipid droplet in single fission yeast cells in vivo by Raman microscopy. He has then developed methods to study single cell biochemistry
utilizing carbon isotopes during his Post-doctorate at Ultimate Spectroscopy and Imaging Laboratory, NCTU. Later he moved to Shimane University, Japan as an
Assistant Professor and has been actively working on medical and biological applications of Raman microspectroscopy.