Rapid, High-Throughput Tracking of Bacterial Motility in 3D via Phase-Contrast Holographic Video Microscopy

Fook Chiong Cheong, Chui Ching Wong, Yunfeng Gao, Mui Hoon Nai, Yidan Cui, Sungsu Park, Linda Kenney, Chwee Teck Lim

Research output: Contribution to journalArticle

13 Scopus citations


Tracking fast-swimming bacteria in three dimensions can be extremely challenging with current optical techniques and a microscopic approach that can rapidly acquire volumetric information is required. Here, we introduce phase-contrast holographic video microscopy as a solution for the simultaneous tracking of multiple fast moving cells in three dimensions. This technique uses interference patterns formed between the scattered and the incident field to infer the three-dimensional (3D) position and size of bacteria. Using this optical approach, motility dynamics of multiple bacteria in three dimensions, such as speed and turn angles, can be obtained within minutes. We demonstrated the feasibility of this method by effectively tracking multiple bacteria species, including Escherichia coli, Agrobacterium tumefaciens, and Pseudomonas aeruginosa. In addition, we combined our fast 3D imaging technique with a microfluidic device to present an example of a drug/chemical assay to study effects on bacterial motility.

Original languageEnglish (US)
Pages (from-to)1248-1256
Number of pages9
JournalBiophysical Journal
Issue number5
StatePublished - Mar 10 2015
Externally publishedYes


ASJC Scopus subject areas

  • Biophysics

Cite this