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Biofilms are complex communities of microorganisms encased in a self-produced extracellular polysaccharide matrix attached to surfaces. These microbial consortia can include single or multiple species, providing enhanced survival benefits by forming organized, multilayered structures.The formation of biofilms occurs through four key stages: attachment, colonization, development, and dispersal.During attachment, free-swimming planktonic cells adhere to a surface, often facilitated by...
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Bacterial and archaeal cells exhibit remarkable diversity in shape and structure, critical in their adaptability and functionality. Among bacteria, the most commonly observed shapes include cocci and bacilli. Cocci are spherical and may exist singly or in groupings such as pairs (diplococci), chains (streptococci), clusters (staphylococci), or tetrads. Bacilli, in contrast, are rod-shaped and can also occur as single cells, in pairs, or chains, depending on their environmental and genetic...
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Coryneform bacteria are gram-positive, aerobic, nonmotile rods that exhibit irregular, club-shaped, or V-shaped arrangements. Their V-shape results from snapping division, where the inner cell wall layer forms the cross-wall, while the outer layer remains intact until it ruptures on one side, causing the daughter cells to bend away.The primary genera are Corynebacterium and Arthrobacter. Corynebacterium includes diverse species, ranging from saprophytes to pathogens like Corynebacterium...
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Related Experiment Video

Updated: Feb 27, 2026

Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry
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Morphologies and phenotypes in Bacillus subtilis biofilms.

Xiaoling Wang1,2, Shuo Meng3, Jingshi Han3

  • 1School of Mechanical Engineering, University of Science and Technology, Beijing, P. R. China. xiaoling@me.ustb.edu.cn.

Journal of Microbiology (Seoul, Korea)
|July 5, 2017
PubMed
Summary
This summary is machine-generated.

Researchers studied Bacillus subtilis biofilm growth by altering substrate stiffness and nutrient availability. Environmental factors can be manipulated to control biofilm development.

Keywords:
Bacillus subtilisbiofilmfluorescencemorphologyphenotype

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Area of Science:

  • Microbiology
  • Biophysics
  • Biotechnology

Background:

  • Bacillus subtilis biofilms are crucial in various environments.
  • Understanding biofilm formation is key to controlling microbial communities.
  • Environmental factors significantly influence bacterial biofilm development.

Purpose of the Study:

  • To investigate Bacillus subtilis biofilm growth under varied substrate stiffness and nutrient conditions.
  • To develop quantitative methods for characterizing biofilm morphology and cell phenotype distribution.
  • To establish methods for tuning biofilm growth through environmental manipulation.

Main Methods:

  • Utilized advanced optical imaging for spatial and temporal tracking of biofilm growth.
  • Developed a quantitative method using the P2A ratio to assess biofilm rim irregularities.
  • Employed Beer-Lambert's law and cross-sectioning for biofilm thickness estimation.
  • Used a triple-fluorescence-labeled B. subtilis strain to analyze cell phenotype distributions (motility, matrix production, sporulation).

Main Results:

  • Demonstrated that substrate stiffness and nutrient levels significantly impact Bacillus subtilis biofilm growth.
  • Successfully quantified biofilm morphology and rim irregularities using the P2A ratio.
  • Established a reliable method for estimating biofilm thickness from transmission images.
  • Identified distinct distributions of B. subtilis cell phenotypes within biofilms under different conditions.

Conclusions:

  • Bacillus subtilis biofilm growth is highly sensitive to environmental parameters like substrate stiffness and nutrient availability.
  • The developed quantitative methods provide novel tools for characterizing biofilm structure and dynamics.
  • Environmental conditions can be modulated to effectively tune and control biofilm development.