Most macromolecules, such as lipids, ribosomes, proteins, plasmids, metabolites, and RNA species move in varying diffusive states with different velocities in a crowded environment ( Mika and Poolman, 2011). The rapid development and implementation of fluorescence microscopy techniques has led to the realization that, even in tiny bacterial cells, key molecular processes can be highly orchestrated and organized to temporarily or statically occur at specific subcellular sites ( Rudner and Losick, 2010 Amster-Choder, 2011 Campos and Jacobs-Wagner, 2013). It includes a critical evaluation of their caveats as well as potential solutions. Here, an overview will be given of fluorescent techniques that can be used to reveal specific RNA molecules inside fixed and living single bacterial cells.
Many of these approaches are useful for the large eukaryotic cells but their adaptation to study RNA, specifically mRNA molecules, in bacterial cells progressed relatively slow. Since then, a plethora of techniques to image RNA molecules with the aid of fluorescence microscopy has emerged. The first methods that allowed observing single mRNA molecules in individual cells were introduced by Bertrand et al. To be able to visualize the abundance and spatiotemporal features of RNAs in bacterial cells would permit obtaining a pivotal understanding of many mechanisms underlying bacterial cell biology.
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