Combining Fluorescence and Bioluminescence Microscopy to Study the Series of Events from Cellular Signal Transduction to Gene Expression

Curr Protoc Cell Biol. 2017 Dec 11:77:4.35.1-4.35.16. doi: 10.1002/cpcb.35.

Abstract

The molecular interactions and translocation of signal transduction factors in individual cells can be imaged by fluorescence microscopy. Alternatively, downstream promoter activity in single cells can be imaged by bioluminescence microscopy. However, the same stimuli can lead to different gene expression responses in individual cells. For this reason, it is desirable to simultaneously image signal transduction and gene expression events in the same cells. Here, we describe a method that combines fluorescence and bioluminescence microscopy to image protein kinase C (PKC) translocation from the cytosol to the plasma membrane and the expression of nuclear factor kappa-light polypeptide B (NF-κB)-regulated genes. © 2017 by John Wiley & Sons, Inc.

Keywords: bioluminescence; fluorescence; gene expression; imaging; signal transduction; single-cell analysis.

MeSH terms

  • Equipment Design
  • Gene Expression
  • Gene Expression Regulation*
  • HeLa Cells
  • Humans
  • Luminescence
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • NF-kappa B / metabolism
  • Optical Imaging / instrumentation
  • Optical Imaging / methods*
  • Protein Kinase C / analysis
  • Protein Kinase C / metabolism
  • Protein Transport
  • Signal Transduction*
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods*

Substances

  • NF-kappa B
  • Protein Kinase C