Multimodal segmentation of dynamic subcellular features using quantitative phase imaging and FRET-based sensors...

Area of Science:

• Cellular biology
• Biophysics
• Quantitative imaging

Background:

• Cellular mechanotransduction research relies on understanding responses to mechanical stimuli.
• Fluorescence microscopy is valuable for molecular sensitivity, but quantitative phase imaging (QPI) for morphology visualization is underutilized.
• Previous work combined QPI with Förster resonance energy transfer (FRET) constructs, necessitating further analysis techniques.

Purpose of the Study:

• To develop novel analysis techniques for QPI to characterize individual cell responses to mechanical stimuli.
• To integrate QPI with FRET-based sensors for multimodal imaging of cellular responses.
• To investigate cellular responses to hypo-osmotic stimulus, focusing on calcium ion flux and intracellular mass reorganization.

Main Methods:

• Developed a QPI-guided cellular segmentation algorithm for characterizing cell responses.
• Employed a multimodal imaging instrument combining QPI and FRET-based calcium ion sensors.
• Examined cellular responses to hypo-osmotic stimulus, mapping intracellular mass reorganization and calcium flux.

Main Results:

• The multimodal imaging approach enabled discrimination of cell responses by localized region.
• Distinct cellular behaviors were observed between different regions and compared to a control group.
• Novel analysis techniques identified cell expansion and region-specific responses in both QPI and FRET modalities.

Conclusions:

• The combination of QPI and FRET sensors provides a powerful tool for studying cellular responses to environmental stimuli.
• QPI offers new avenues for mechanotransduction research by complementing molecularly sensitive FRET sensors.
• This approach allows for detailed characterization of cell expansion and localized responses.