Quantitative dynamic cellular imaging based on 3D unwrapped optically computed phase microscopy.

Area of Science:

• Biophysics
• Optical Imaging
• Microscopy

Background:

• Continuous observation of dynamic phenomena is crucial for understanding biological processes.
• Quantitative phase microscopy (QPM) offers label-free imaging of cellular dynamics.
• Phase wrapping artifacts in QPM hinder accurate quantification of sample dynamics.

Purpose of the Study:

• To develop and validate a novel 3D phase unwrapping method for QPM.
• To address phase wrapping artifacts that limit the quantification of dynamic phenomena.
• To enable accurate characterization of nanoscale motion and live cell dynamics using QPM.

Main Methods:

• Development of a 3D phase unwrapping algorithm exploiting spatiotemporal data correlation.
• Validation using simulated data to assess algorithm performance.
• Experimental validation using a piezo transducer (PZT) for nanoscale motion and live cell detachment imaging.

Main Results:

• The 3D phase unwrapping method successfully removes phase wrapping artifacts.
• Quantitative phase imaging accurately characterizes sample dynamics.
• Calculated displacements of PZT-actuated samples match known motion.
• Observed increasing optical path length in detaching cells due to dry mass concentration.

Conclusions:

• The developed 3D phase unwrapping method significantly improves quantitative phase imaging.
• This technique enables accurate, artifact-free characterization of dynamic phenomena at the nanoscale.
• The method provides valuable insights into cellular processes like detachment through precise dry mass and optical path length measurements.