Focusing light on infection in four dimensions.
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.
Roux
Pascal
P
Plate-forme d'Imagerie Dynamique (PFID), Institut Pasteur, 25-28 rue du Dr Roux, 75015 Paris, France.
Münter
Sylvia
S
Frischknecht
Freddy
F
Herbomel
Philippe
P
Shorte
Spencer L
SL
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D005453:Fluorescence; D006790:Host-Parasite Interactions; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D007239:Infections / Q000382:microbiology / Q000469:parasitology / Q000821:virology; D008027:Light; D008853:Microscopy / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods