Journal of Microbial & Biochemical Technology

Imaging yeast cell wall architecture and studying the effect of antifungal drug Terbinafine at the single cell level by Raman microscopy

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:

Fungal cell wall is a dynamic organelle that plays a vital role, particularly in cellular growth, elongation and division. Any modification or disruption of the wall leads to lysis and cell death, hence serving an excellent target for anti-fungal drugs. Chemically, the cell wall architecture in fungi is complex comprising mainly polysaccharides (glucan, mannan and chitin) and a small proportion of glycoproteins. Traditionally, electron microscopy and biochemical extraction methods were employed while recently immunocytochemical analysis is used to understand its structure. The former lacks chemical specificity requiring genetically modified cells to study different structures in detail while the later involves development of fluorescent monoclonal antibodies specific to glycosidic linkages among cell wall polysaccharides. Hence we aim to develop a label-free method based on confocal Raman microscopy to visualize distribution of various polysaccharide components of fungal cell and spore wall. Fission yeast Schizosaccharomyces pombe is used as a model to demonstrate our method. First, space-resolved Raman spectra from lipid droplets, cytoplasm and cell wall were obtained to identify marker bands for individual components followed by imaging. Then, by employing multivariate curve resolution (MCR) analysis, we successfully separated Raman spectra of several pure bio-macromolecular components. We then studied pharmacokinetics and pharmacodynamics of antifungal drug terbinafine at the single cell level in yeast model. We believe that our method will help in understanding the complex fungal spore wall architecture and eventually lead to advancements in drug discovery and development in the future.

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 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