Bacteria Growth

Introduction

Understanding bacteria is crucial given their widespread presence in various environments, vital roles in ecosystems, and substantial impact on human health. Traditional methods like agar plate culture and limiting dilution are limited: agar plates favour fast-growing microorganisms, while the limiting dilution method is labor-intensive and slow. LiveDrop instruments revolutionise bacterial studies by enabling individual sorting in picodroplets, overcoming these challenges. Diversity and complexity of the isolated microbes (Terekhov et al 2020, Mahler2020, Watterson et al 2020).

Fundamentals

‍The study of bacteria remains a vast field with a significant portion still unexplored; it is estimated that 99% of bacterial species are yet to be identified. Given their substantial impact on ecosystems and human health, expanding our understanding of these microbes is imperative. Traditionally, bacteria are cultured on agar plates to facilitate their study during growth. However, several challenges hinder bacterial research, including the overgrowth of fast-growing strains, which complicates the study of slower-growing strains and their properties.

LiveDrop Technology

Utilising picodroplets, LiveDrop instruments enable the encapsulation and separation of bacteria. Bacteria can be sorted individually or in groups (see Figure 1). Sorting bacteria individually facilitates the study of single strain physiology or its response to various compounds. Sorting them in groups enables scientists to gather data on interactions between different strains.

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Figure 1: Illustration of bacteria sorting with Modaflow.Bacteria encapsulation in picodroplets using ModaFlow. Various conditions possible: one droplet, one bacterium, or one droplet with multiple bacteria.

Results

Bacteria sorted into droplets have the capability to grow and form new colonies. As depicted in Figure 2, visual observation of bacteria is feasible for several days post-sorting. Even after 96 hours, droplets containing viable bacterial colonies remain visible. Additionally, droplets can be transferred to agar plates to extend the observation period.

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Figure 2: The E. coli strain is compartmentalised into 30 to 40 pL droplets using the ModaFlow instrument, collected in a 1.5 ml tube, and incubated without agitation at 37°C. To monitor bacterial growth in droplets, 10 mL of the emulsion is transferred into a DVD device.(a) Image of the emulsion immediately after droplet production. The red circle indicates a droplet containing 2 bacteria, and the blue circle indicates another droplet containing 2 bacteria. Images (b), (c), and (d) depict bacterial growth after 24, 48, and 96 hours of incubation, respectively.

Conclusion

LiveDrop's instruments, by isolating bacteria in picodroplets, facilitate extended observation of bacterial cultures spanning multiple days. This allows for visual examination of bacterial behaviours, providing researchers with a convenient means to investigate strains characterised by slow growth rates.

The team

LiveDrop colleagues

FAN

OneFlow

Microbiology