Journal of Microbial & Biochemical Technology

Stable isotope probing coupled Raman microscopy: An efficient way to study single cell biochemistry

International Conference and Summit on Industrial & Pharmaceutical Microbiology

October 17-18, 2016 Kuala Lumpur, Malaysia

Hemanth Nag Noothalapati Venkata

Shimane University, Japan

Scientific Tracks Abstracts: J Microb Biochem Technol

Abstract:

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 stableisotope (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 13C 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.

Email: nvhnag@life.shimane-u.ac.jp