Three-dimensional imaging flow cytometry through light-sheet fluorescence microscopy

Cytometry A. 2017 Feb;91(2):144-151. doi: 10.1002/cyto.a.23046. Epub 2017 Jan 11.

Abstract

Flow cytometry is the tool of choice for high-speed acquisition and analysis of large cell populations, with the tradeoff of lacking intracellular spatial information. Although in the last decades flow cytometry systems that can actually acquire two-dimensional spatial information were developed, some of the limitations remained though, namely constrains related to sample size and lack of depth or dynamic information. The combination of fluidics and light-sheet illumination has the potential to address these limitations. By having cells travelling with the flowing sheath one can, in a controlled fashion, force them at constant speed through the light-sheet enabling the synchronized acquisition of several optical sections, that is, three-dimensional imaging. This approach has already been used for imaging cellular spheroids, plankton, and zebra-fish embryos. In this review, we discuss the known solutions and standing challenges of performing three-dimensional high-throughput imaging of multicellular biological models using fluidics, while retaining cell and organelle-level resolution. © 2017 International Society for Advancement of Cytometry.

Keywords: high throughput microscopy; imaging flow cytometry; light sheet fluorescence microscopy.

Publication types

  • Review
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Flow Cytometry / methods*
  • High-Throughput Screening Assays
  • Imaging, Three-Dimensional / methods*
  • Microscopy, Fluorescence / methods*
  • Plankton / ultrastructure
  • Spheroids, Cellular / ultrastructure
  • Zebrafish