Focusing light on infection in four dimensions

• Published in: Cellular microbiology Vol. 6; no. 4; pp. 333 - 343
• Main Authors: Roux, Pascal, Münter, Sylvia, Frischknecht, Freddy, Herbomel, Philippe, Shorte, Spencer L
• Format: Journal Article
• Language: English
• Published: England 01.04.2004
• Subjects: Animals; Cell Line; Cells, Cultured; Fluorescence; Host-Parasite Interactions; Humans; Image Processing, Computer-Assisted - methods; Imaging, Three-Dimensional - instrumentation; Imaging, Three-Dimensional - methods; Infection - microbiology; Infection - parasitology; Infection - virology; Light; Microscopy - methods; Microscopy, Confocal - methods; Microscopy, Fluorescence - methods
• ISSN: 1462-5814; 1462-5822
• DOI: 10.1111/j.1462-5822.2004.00374.x

The fusion of cell biology with microbiology has bred a new discipline, cellular microbiology, in which the primary aim is to understand host-pathogen interactions at a tissue, cellular and molecular level. In this context, we require techniques allowing us to probe infection in situ and extrapolate quantitative information on its spatiotemporal dynamics. To these ends, fluorescent light-based imaging techniques offer a powerful tool, and the state-of-the-art is defined by paradigms using so-called multidimensional (multi-D) imaging microscopy. Multi-D imaging aims to visualize and quantify biological events through time and space and, more specifically, refers to combinations of: three (3D, volume), four (4D, time) and five (5D, multiwavelength)-dimensional recordings. Successful multi-D imaging depends upon understanding the available technologies and their limitations. This is especially true in the field of microbiology where visualization of infectious/pathogenic activities inside living host systems presents particular technical challenges. Thus, as multi-D imaging rapidly becomes a common bench tool to the cellular microbiologist, this review provides the new user with some of the necessary technical insight required to get the best from these methods.