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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: Jun 9, 2025

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

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Published on: April 7, 2014

Quantitative Measurements of Red Blood Cell Indices Using Spectroscopic Differential Phase-Contrast Microscopy.

Taegyun Moon¹, Andrew Heegeon Yang¹, Seungri Song¹

¹Department of Mechanical Engineering, Yonsei University, Seodaemoon-gu, Seoul 03722, Republic of Korea.

Chemical & Biomedical Imaging

|October 30, 2024

Summary

This summary is machine-generated.

Spectroscopic differential phase-contrast (sDPC) microscopy accurately measures red blood cell (RBC) indices using minimal sample volume. This advanced imaging technique offers a precise alternative to conventional methods for diagnosing blood disorders.

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Last Updated: Jun 9, 2025

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

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Immunostaining-Based Detection of Dynamic Alterations in Red Blood Cell Proteins

Immunostaining-Based Detection of Dynamic Alterations in Red Blood Cell Proteins

Published on: March 17, 2023

Area of Science:

Biomedical Optics
Hematology
Computational Imaging

Background:

Red blood cell (RBC) indices are crucial for diagnosing blood diseases.
Conventional analyzers require large blood volumes and provide only averaged data.
Existing quantitative phase imaging (QPI) methods have limitations like complex setups and speckle noise.

Purpose of the Study:

To introduce spectroscopic differential phase-contrast (sDPC) microscopy for measuring RBC indices.
To overcome limitations of conventional methods and coherent QPI.
To enable accurate, single-cell level analysis of RBC morphology and chemistry.

Main Methods:

Utilized sDPC microscopy, a computational imaging technique.
Generated color-dependent phase images with high resolution and reduced speckle.
Applied computational algorithms to extract RBC indices from spectroscopic phase images.

Main Results:

Achieved high-accuracy measurements of multiple RBC indices (e.g., MCHC, MCV, MCH, RDW, HCT, Hb, RBC count).
Demonstrated errors less than 7% compared to a clinical flow cytometry analyzer.
Validated clinical utility by comparing sDPC results with a standard analyzer in control and anemic groups.

Conclusions:

sDPC microscopy is a viable platform for accurate RBC index measurement.
The method offers advantages in spatial resolution and noise reduction over coherent QPI.
sDPC shows potential for improved diagnostics in hematology, especially for single-cell analysis.