Abstract
Bacteria in nature live in complex communities with multiple cell types and spatially-dependent interactions. Studying cells in well-mixed environments such as shaking culture tubes or flasks cannot capture these spatial dynamics, but cells growing in full-fledged biofilms are difficult to observe in real time. We present here a protocol for observing time-resolved, multi-species interactions at single-cell resolution. The protocol involves growing bacterial cells in a near monolayer in a microfluidic device. As a demonstration, we describe in particular observing the dynamic interactions between E. coli and Acinetobacter baylyi. In this case, the protocol is capable of observing both contact-dependent lysis of E. coli by A. baylyi via the Type VI Secretion System (T6SS) and subsequent functional horizontal gene transfer (HGT) of genes from E. coli to A. baylyi.
Highlights
[Background] Spatially-dependent interactions between different species of bacteria likely occur ubiquitously in nature, but they can be difficult to observe
One example is enhancement of horizontal gene transfer (HGT) by contact-dependent, in situ lysis of a prey cell, which serves as DNA donor, by a naturally competent, predatory, DNA recipient cell
This was only recently observed in Gram-negative bacteria, but it has already been seen in multiple species, and it is thought to be a relatively widespread phenomenon (Borgeaud et al, 2015; Cooper et al, 2017; Veening and Blokesch, 2017; Ringel et al, 2017)
Summary
[Background] Spatially-dependent interactions between different species of bacteria likely occur ubiquitously in nature, but they can be difficult to observe. 1. Glass coverslips, #1.5 (Fisher Scientific, catalog number: 12-530F) 2. 1.5 ml microcentrifuge tubes (Fisher Scientific, catalog number: 05-408-129) 12.
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