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A novel time-lapse imaging method for studying developing bacterial biofilms

Futo, Momir; Široki, Tin; Koska, Sara; Čorak, Nina; Tušar, Anja; Domazet-Lošo, Mirjana; Domazet-Lošo, Tomislav (2022) A novel time-lapse imaging method for studying developing bacterial biofilms. Scientific Reports, 12 . ISSN 2045-2322

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In nature, bacteria prevailingly reside in the form of biofilms. These elaborately organized surface‑bound assemblages of bacterial cells show numerous features of multicellular organization. We recently showed that biofilm growth is a true developmental process, which resembles developmental processes in multicellular eukaryotes. To study the biofilm growth in a fashion of eukaryotic ontogeny, it is essential to define dynamics and critical transitional phases of this process. The first step in this endeavor is to record the gross morphological changes of biofilm ontogeny under standardized conditions. This visual information is instrumental in guiding the sampling strategy for the later omics analyses of biofilm ontogeny. However, none of the currently available visualizations methods is specifically tailored for recording gross morphology across the whole biofilm development. To address this void, here we present an affordable Arduino‑based approach for time‑lapse visualization of complete biofilm ontogeny using bright field stereomicroscopy with episcopic illumination. The major challenge in recording biofilm development on the air–solid interphase is water condensation, which compromises filming directly through the lid of a Petri dish. To overcome these trade‑offs, we developed an Arduino microcontroller setup which synchronizes a robotic arm, responsible for opening and closing the Petri dish lid, with the activity of a stereomicroscope‑mounted camera and lighting conditions. We placed this setup into a microbiological incubator that maintains temperature and humidity during the biofilm growth. As a proof‑of‑principle, we recorded biofilm development of five Bacillus subtilis strains that show different morphological and developmental dynamics.

Item Type: Article
Uncontrolled Keywords: bacterail biofilms ; time‑lapse visualization ; Arduino ; stereomicroscopy
Subjects: NATURAL SCIENCES > Biology
INTERDISCIPLINARY AREAS OF KNOWLEDGE > Biotechnology in Biomedicine (natural science, biomedicine and healthcare, bioethics area
Divisions: Division of Molecular Biology
Project titleProject leaderProject codeProject type
Filostratigrafija nastanka i gubitka genaDomazet-Lošo, TomislavIP-2016-06-5924HRZZ
Depositing User: Tomislav Domazet-Lošo
Date Deposited: 03 Apr 2023 10:46
DOI: 10.1038/s41598-022-24431-y

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