Nanoscale imaging of host-pathogen interactions

Thursday, October 11, 2012 — Poster Session III
10:00 a.m. – Noon	Natcher Conference Center, Building 45	NCI	IMAG-22

* FARE Award Winner

Authors

• K. Narayan
• G.E. Murphy
• T.P. Do
• L. Hartnell
• B. Lowekamp
• S. Subramaniam

Abstract

Understanding the hierarchical organization of molecules, multi-protein assemblies, organelles and networks within the interior of a eukaryotic cell is a challenge of fundamental interest in cell biology. We have been developing a battery of tools to provide nanoscale 3D structural and chemical information of biological specimens to complement advances in fluorescence microscopy that allow cellular imaging at sub-micron resolutions. Examples we present here include (i) Use of Ion Abrasion Scanning Electron Microscopy, IA-SEM, also known as Focused Ion Beam, FIB-SEM, to obtain 3D images of resin-embedded biological specimens by iteratively recording SEM images of the specimen freshly exposed by milling or “slicing” by the abrasive focused ion beam. (ii) Imaging T cell-HIV interactions in 3D with correlative imaging using live confocal microscopy coupled with IA-SEM. (iii) Mapping the chemical composition of unstained cells using a combination of secondary ion mass spectrometry and focused ion beam milling, and finally (iv) Nanometer resolution 3D compositional mapping of portions of unstained cells by atom probe tomography, a novel method where controlled emissions of ions can be induced from specimens milled to the requisite needle shape by the focused ion beam.

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