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Phase Contrast and Differential Interference Contrast Microscopy

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Updated: Jun 12, 2026

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

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

Published on: April 7, 2014

Sample-less calibration of the differential interference contrast microscope.

Shalin B Mehta¹, Colin J R Sheppard

¹1Optical Bioimaging Lab, Division of Bioengineering, National University of Singapore,Block-E3A, #7-10, 7 Engineering Drive 1, Singapore 117574.

|May 22, 2010

Summary

This summary is machine-generated.

Accurate calibration of differential interference contrast (DIC) microscope shear and bias is crucial for image analysis. This study introduces a simple, specimen-free method using interference fringes for precise DIC parameter measurement.

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Performing Spectroscopy on Plasmonic Nanoparticles with Transmission-Based Nomarski-Type Differential Interference Contrast Microscopy

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Published on: June 5, 2019

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Last Updated: Jun 12, 2026

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

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Published on: April 7, 2014

Performing Spectroscopy on Plasmonic Nanoparticles with Transmission-Based Nomarski-Type Differential Interference Contrast Microscopy

Performing Spectroscopy on Plasmonic Nanoparticles with Transmission-Based Nomarski-Type Differential Interference Contrast Microscopy

Published on: June 5, 2019

Area of Science:

Optical microscopy
Image analysis
Metrology

Background:

Differential interference contrast (DIC) microscopy is essential for visualizing unstained biological specimens.
Accurate measurement of DIC microscope parameters, shear and bias, is critical for quantitative image analysis.
Existing methods for DIC calibration often rely on approximate measurements or complex setups.

Purpose of the Study:

To present a novel, accurate, and simple method for measuring DIC microscope shear and bias.
To eliminate the need for calibrated specimens in DIC parameter determination.
To provide a clear explanation of the underlying optical principles involving Nomarski prisms.

Main Methods:

Analysis of interference fringes in the back focal plane of the objective lens.
Image analysis techniques applied to the fringe patterns.
Theoretical description of Nomarski prism properties and their role in interference.

Main Results:

A precise and reliable method for measuring DIC shear and bias was developed.
The method demonstrated accuracy without requiring external calibrated specimens.
The technique simplifies the calibration process for DIC microscopes.

Conclusions:

The presented method offers an accurate and straightforward approach to DIC microscope calibration.
This technique enhances the reliability of quantitative image analysis in DIC microscopy.
The findings contribute to improved optical metrology in microscopy.