Focused Ion Beam (FIB) is a novel technique that allows easy target cell selection, fast operation, high resolution, 3D imaging and sample manipulation during imaging. The FIB technique of microscopy and nanomachining, that is widely spread in semiconductor technology, is reshuffled to open new horizons in the field oflife sciences at cellular and subcellular level. FIB is a source ofions which can be precisely oriented and focused on the sample, supported also by an electron source. The resolution can be as low as I 5nm. Two different operation modes are available: "cutting" and "etching" operations of very high precision and at the same time the resulting secondary ions and electrons provide follow up sample imaging. FIB tomography ability has been tested to provide information on cell characterisation, cell division time sequence, view of inner structures and investigation of membrane structural properties. We compare the performances and the advantages of different high-resolution microscopy techniques: Soft X-ray Contact Microscopy (SXCM), Focused Ion Beam (FIB) and Transmission Electron Microscopy (TEM). These have been used to image Saccharoniyces cerevisiae yeast cells, a rather well known sample of great biological interest. TEM is an established technique used as a benchmark for comparison.

Date Published: 16 April 2001
PDF: 13 pages
Proc. SPIE 4261, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VIII, (16 April 2001); doi: 10.1117/12.424523

Published in SPIE Proceedings Vol. 4261:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VIII
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson, Editor(s)