Characterization of bacterial spores using Nano-Secondary Ion Mass Spectrometry (NanoSIMS)

ANYL 315

Sutapa Ghosal, ghosal2@llnl.gov1, Stewart J. Fallon1, Terrance Leighton2, Katie Wheeler2, I. D. Hutcheon, N/A1, and Peter K. Weber1. (1) Lawrence Livermore National Laboratory, L-231, P.O. Box 808, 7000 East Ave., Livermore, CA 94550, (2) Children’s Hospital Oakland Research Institute
Bacterial spores are elementally zoned at the nanometer scale. This zonation may be controlled by spore physiology, physical factors, and elemental diffusion. Here we present a recently developed nanometer-scale secondary ion mass spectrometry (NanoSIMS) technique that allows the direct visualization and quantification of elemental concentration gradients within spores. By using NanoSIMS depth profile analysis together with sample preparation techniques such as focused ion beam (FIB) sectioning, we are able to probe the three dimensional elemental distribution within individual Bacillus thuringiensis israelensis (Bti) spores with nanometer scale resolution (~10 nm depth and 50 nm lateral). Our results show the expected distributions for physiologically controlled elements (Ca, P) and provide baseline data other elements (e.g., Li, F, Cl, S). We also demonstrate cation and anion mobility in spores under hydrous conditions. Our results suggest a permeation mechanism by which elements diffuse into and out of the spore along hydration pathways on rather short time scales. Additional studies are in progress to define the rates and mechanisms controlling ion mobility in spores.

This work was performed under the auspices of the U.S. Department of Energy by the University of California, Lawrence Livermore National Laboratory under Contract W-7405-Eng-4.