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Phase Contrast and Differential Interference Contrast Microscopy

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In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...

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Updated: Apr 22, 2026

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope

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Quantitative Phase Microscopy: how to make phase data meaningful.

Goldie Goldstein¹, Katherine Creath²

¹4D Technology Corporation, Tucson AZ 85706.

Proceedings of Spie--The International Society for Optical Engineering

|October 14, 2014

Summary

Quantitative phase microscopy (QPM) enables dynamic cell morphology analysis. Advanced processing allows consistent phase unwrapping and accurate tracking of cellular features like vesicles.

Keywords:

cell dynamics cellular imaging interference microscopy optical thickness measurement phase imaging polarization interferometry

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Last Updated: Apr 22, 2026

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Area of Science:

Biophysics
Cell Biology
Optical Imaging

Background:

Quantitative phase microscopy (QPM) hardware and image processing have advanced, enabling superior phase data acquisition.
Recent research links QPM data and metrics to cell morphology, but challenges remain in phase unwrapping and temporal correction.

Purpose of the Study:

To develop consistent phase unwrapping and temporal correction methods for QPM data.
To enable accurate characterization and dynamic tracking of biological samples, such as myocytes and myoblasts.

Main Methods:

Utilized advanced image processing techniques for quantitative phase microscopy.
Implemented an object characterization algorithm for automatic removal of background shape, mean value fluctuations, and residual tilts.
Developed a temporal unwrapping procedure to correct for temporal discrepancies.

Main Results:

Achieved consistent phase unwrapping and accurate dimensional measurements.
Enabled dynamic tracking of cell size, texture, and optical volume.
Successfully identified intracellular objects like vesicles within myoblasts, even when co-located.

Conclusions:

QPM offers a label-free method for characterizing living cells and their morphology in dynamic environments.
Consistent and automated data processing is critical for QPM to be widely adopted and integrated with other imaging modalities.
Connecting QPM measurements to biological function is key for broader scientific community impact.