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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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Related Experiment Video

Updated: Sep 23, 2025

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

Time-lapse imaging using dual-color coded quantitative differential phase contrast microscopy.

Ying-Ju Chen^1,2, Yu-Zi Lin², Sunil Vyas²

¹National Taiwan University, Department of Biomedical Engineering, Taiwan.

Journal of Biomedical Optics

|May 17, 2022

Summary

This summary is machine-generated.

Quantitative differential phase contrast (qDPC) microscopy now achieves high-contrast live cell imaging with just two measurements. This advancement significantly reduces imaging time and motion artifacts, enabling detailed observation of cellular dynamics.

Keywords:

phase contrast quantitative phase imaging time-lapse imaging

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Last Updated: Sep 23, 2025

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Simultaneously Capturing Real-time Images in Two Emission Channels Using a Dual Camera Emission Splitting System: Applications to Cell Adhesion

Simultaneously Capturing Real-time Images in Two Emission Channels Using a Dual Camera Emission Splitting System: Applications to Cell Adhesion

Published on: September 4, 2013

Area of Science:

Biomedical Optics
Microscopy Techniques
Cellular Imaging

Background:

Quantitative differential phase contrast (qDPC) microscopy uses asymmetric illumination and multiple intensity measurements to enhance phase contrast.
Live cell imaging in qDPC is challenged by motion artifacts and lengthy acquisition times due to numerous intensity measurements.
Reducing the number of intensity measurements is crucial for improving live imaging quality and speed in qDPC.

Purpose of the Study:

To achieve high-contrast, isotropic qDPC images using only two intensity measurements.
To enable time-lapse imaging of biological samples with enhanced speed and reduced artifacts.
To demonstrate the efficacy of a novel pupil design for efficient qDPC phase retrieval.

Main Methods:

A dual-color linear-gradient pupil was designed to provide asymmetric illumination and achieve a circularly symmetric transfer function.
This dual-color pupil design reduces the required number of frames for phase retrieval, enabling faster image acquisition.
The system's performance was validated using standard microlens arrays and time-lapse imaging of rat astrocytes.

Main Results:

Isotropic phase contrast response was achieved with only two-axis measurements using the dual-color linear-gradient pupil.
The reduced frame acquisition significantly increased imaging speed compared to conventional qDPC systems.
Time-lapse imaging of rat astrocytes revealed detailed morphology and dynamic changes, including apoptosis and migration.

Conclusions:

Dual-color linear-gradient pupils offer superior performance in qDPC compared to half-circle and vortex pupils.
Achieving isotropic phase transfer function with minimal measurements (two-axis) enhances qDPC applicability.
The developed qDPC method facilitates faster, high-quality live cell imaging, enabling detailed observation of cellular processes.