Capturing cell morphology dynamics with high temporal resolution using single-shot quantitative phase gradient...

Area of Science:

• Biomedical Optics
• Cell Biology
• Microscopy

Background:

• Label-free quantitative phase imaging (QPI) offers non-perturbing measurement of cellular dynamics.
• Developing faster and more sensitive instrumentation is crucial for capturing rapid cellular events.
• Single-shot quantitative phase gradient microscopy (ss-QPGM) enables simultaneous acquisition of polarization components for phase reconstruction.

Purpose of the Study:

• To characterize a fast, single-shot quantitative phase gradient microscopy (ss-QPGM) approach.
• To record morphological changes in single-cell phase images.
• To correlate these morphological changes with biochemical indicators of cell death using fluorescence imaging.

Main Methods:

• Implemented a computationally efficient least squares algorithm for real-time, video-rate imaging (up to 30 fps).
• Utilized ss-QPGM to acquire phase images and concurrently acquired fluorescence images of biochemically labeled cells.
• Validated phase measurement accuracy using a USAF1951 pattern phase target and calibrated sample thicknesses.

Main Results:

• Achieved a spatial resolution of 0.8 µm and accurate phase retrieval (R² > 0.99).
• Demonstrated real-time, video-rate (30 fps) ss-QPGM for live-cell imaging.
• Observed dynamic intracellular and morphological changes during nutrient deprivation and drug-induced cell death, correlating with necrosis.

Conclusions:

• Label-free ss-QPGM provides high-temporal resolution and high spatial fidelity for live-cell imaging.
• The technique effectively monitors dynamic cellular changes during induced cell death.
• ss-QPGM shows significant promise for studying live-cell dynamics with minimal perturbation.