Molecular and biophysical analysis of apoptosis using a combined quantitative phase imaging and fluorescence resonance energy transfer microscope

J Biophotonics. 2018 Dec;11(12):e201800126. doi: 10.1002/jbio.201800126. Epub 2018 Aug 16.

Abstract

Apoptotic mechanisms are often dysregulated in cancerous phenotypes. Additionally, many anticancer treatments induce apoptosis and necrosis, and the monitoring of this apoptotic activity can allow researchers to identify therapeutic efficiency. Here, we introduce a microscope which combines quantitative phase imaging (QPI) with the ability to detect molecular events via fluorescence (or Förster) resonance energy transfer (FRET). The system was applied to study cells undergoing apoptosis to correlate the onset of apoptotic enzyme activity as observed using a FRET-based apoptosis sensor with whole cell morphological changes analyzed via QPI. The QPI data showed changes in cell disorder strength during the initiation of apoptotic enzymatic activity.

Keywords: Förster resonance energy transfer; apoptosis; disorder strength; quantitative phase imaging.

Publication types

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

MeSH terms

  • Apoptosis*
  • Biophysics / methods*
  • Caspase 3 / metabolism
  • Enzyme Activation
  • Fluorescence Resonance Energy Transfer*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*

Substances

  • Caspase 3