Application of Single-Cell Bioluminescent Imaging to Monitor Circadian Rhythms of Individual Plant Cells

Methods Mol Biol. 2020:2081:231-242. doi: 10.1007/978-1-4939-9940-8_17.

Abstract

The bioluminescent reporter system is a powerful tool for the long-term monitoring of gene expression because of its noninvasive nature. Furthermore, in combination with high-sensitive imaging technology, spatiotemporal analysis on regulation and heterogeneity in gene expression is possible. We developed a single-cell bioluminescent imaging system for plants through a transient gene transfection by particle bombardment. By applying this system to a duckweed species, we succeeded in monitoring circadian rhythms of individual cells in an intact plant for over a week. Here we describe methods for gene transfection by particle bombardment and single-cell bioluminescence monitoring by a high-sensitive camera. This technique provides a platform for characterizing gene expression patterns of individual cells in the same tissue.

Keywords: Bioluminescence; Circadian rhythm; Duckweed; Gene expression; Single cell.

MeSH terms

  • Araceae / physiology
  • Circadian Rhythm / physiology*
  • DNA
  • Data Analysis
  • Luminescent Measurements / methods*
  • Molecular Imaging* / methods
  • Single-Cell Analysis* / methods
  • Transfection / methods

Substances

  • DNA