High-speed imaging of amoeboid movements using light-sheet microscopy

PLoS One. 2012;7(12):e50846. doi: 10.1371/journal.pone.0050846. Epub 2012 Dec 5.

Abstract

Light-sheet microscopy has been developed as a powerful tool for live imaging in biological studies. The efficient illumination of specimens using light-sheet microscopy makes it highly amenable to high-speed imaging. We therefore applied this technology to the observation of amoeboid movements, which are too rapid to capture with conventional microscopy. To simplify the setup of the optical system, we utilized the illumination optics from a conventional confocal laser scanning microscope. Using this set-up we achieved high-speed imaging of amoeboid movements. Three-dimensional images were captured at the recording rate of 40 frames/s and clearly outlined the fine structures of fluorescent-labeled amoeboid cellular membranes. The quality of images obtained by our system was sufficient for subsequent quantitative analysis for dynamics of amoeboid movements. This study demonstrates the application of light-sheet microscopy for high-speed imaging of biological specimens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amoeba / physiology*
  • Cell Surface Extensions / physiology
  • Imaging, Three-Dimensional / methods*
  • Light*
  • Microscopy / methods*
  • Movement / physiology*
  • Time Factors

Grant support

This work was supported by the Human Frontier Science Program (HFSP, http://www.hfsp.org/), the Ministry of Education, Culture, Sports, Science and Technology (MEXT, http://www.mext.go.jp/english/), the Japan Society for the Promotion of Science (JSPS, http://www.jsps.go.jp/english/index.html) and CREST (http://www.jst.go.jp/kisoken/crest/en/index.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.