Live-cell fluorescence microscopy to investigate subcellular protein localization and cell morphology changes in bacteria

Robert S. Brzozowski, Maria L. White, Prahathees J. Eswara

Research output: Contribution to journalArticlepeer-review

Abstract

Investigations of factors influencing cell division and cell shape in bacteria are commonly performed in conjunction with high-resolution fluorescence microscopy as observations made at a population level may not truly reflect what occurs at a single cell level. Live-cell timelapse microscopy allows investigators to monitor the changes in cell division or cell morphology which provide valuable insights regarding subcellular localization of proteins and timing of gene expression, as it happens, to potentially aid in answering important biological questions. Here, we describe our protocol to monitor phenotypic changes in Bacillus subtilis and Staphylococcus aureus using a high-resolution deconvolution microscope. The objective of this report is to provide a simple and clear protocol that can be adopted by other investigators interested in conducting fluorescence microscopy experiments to study different biological processes in bacteria as well as other organisms.

Original languageEnglish
Article numbere59905
JournalJournal of Visualized Experiments
Volume2019
Issue number153
DOIs
StatePublished - Nov 2019

Keywords

  • Bacillus subtilis
  • Deconvolution microscopy
  • Fluorescence microscopy
  • FtsZ
  • GFP
  • GpsB
  • Immunology and Infection
  • Issue 153
  • PC190723
  • Protein localization
  • Staphylococcus aureus
  • Timelapse microscopy

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