3D Printing Bacteria to Study Motility and Growth in Complex 3D Porous Media

R. Kōnane Bay, Anna M. Hancock, Arabella S. Dill-Macky, Hao Nghi Luu, Sujit S. Datta

Research output: Contribution to journalArticlepeer-review

Abstract

Bacteria are ubiquitous in complex three-dimensional (3D) porous environments, such as biological tissues and gels, and subsurface soils and sediments. However, the majority of previous work has focused on studies of cells in bulk liquids or at flat surfaces, which do not fully recapitulate the complexity of many natural bacterial habitats. Here, this gap in knowledge is addressed by describing the development of a method to 3D-print dense colonies of bacteria into jammed granular hydrogel matrices. These matrices have tunable pore sizes and mechanical properties; they physically confine the cells, thus supporting them in 3D. They are optically transparent, allowing for direct visualization of bacterial spreading through their surroundings using imaging. As a proof of this principle, here, the capability of this protocol is demonstrated by 3D printing and imaging non-motile and motile Vibro cholerae, as well as non-motile Escherichia coli, in jammed granular hydrogel matrices with varying interstitial pore sizes.

Original languageEnglish (US)
Article numbere66166
JournalJournal of Visualized Experiments
Volume2024
Issue number203
DOIs
StatePublished - 2024

All Science Journal Classification (ASJC) codes

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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