Molecular and biophysical analysis of apoptosis using a combined quantitative phase imaging and fluorescence...

Area of Science:

• Biophysics
• Cell Biology
• Cancer Research

Background:

• Apoptosis dysregulation is common in cancer.
• Anticancer therapies often induce apoptosis, making its monitoring crucial for evaluating treatment efficacy.

Purpose of the Study:

• To develop and apply a novel microscopy technique for simultaneous monitoring of molecular events and morphological changes during apoptosis.
• To correlate early apoptotic enzyme activity with cellular structural alterations.

Main Methods:

• Integration of quantitative phase imaging (QPI) for label-free morphological analysis.
• Incorporation of fluorescence or Förster resonance energy transfer (FRET) for detecting molecular events, specifically apoptotic enzyme activity.
• Application of the combined system to study cells undergoing apoptosis.

Main Results:

• The developed microscope successfully combined QPI and FRET capabilities.
• Correlated the onset of apoptotic enzyme activity (via FRET) with whole-cell morphological changes (via QPI).
• Observed significant changes in cell disorder strength during the initiation of apoptotic enzymatic activity.

Conclusions:

• The novel QPI-FRET microscope enables simultaneous monitoring of cellular morphology and molecular events during apoptosis.
• This integrated approach provides a powerful tool for assessing anticancer therapeutic efficiency by linking molecular signaling to observable cell changes.