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Updated: Jun 27, 2025

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

Quantitative phase imaging by gradient retardance optical microscopy.

Jinming Zhang¹, Mirsaeid Sarollahi¹, Shirley Luckhart^2,3

¹Department of Physics, University of Idaho, 875 Perimeter Drive, Moscow, ID, 83844, USA.

Scientific Reports

|April 28, 2024

Summary

This summary is machine-generated.

Gradient Retardance Optical Microscopy (GROM) enables quantitative phase imaging (QPI) for both thin and thick biological samples. This innovation enhances imaging contrast and cellular analysis in scattering environments.

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Area of Science:

Biophysics
Optical Microscopy
Cellular Imaging

Background:

Quantitative phase imaging (QPI) is crucial for measuring cellular metabolism metrics like dry mass and density.
QPI is limited in optically thick specimens due to scattering, which reduces contrast and increases background noise.

Purpose of the Study:

To introduce Gradient Retardance Optical Microscopy (GROM) for QPI of both thin and thick biological samples.
To adapt standard Differential Interference Contrast (DIC) microscopes into QPI platforms.

Main Methods:

GROM utilizes structured illumination interferometry principles.
A liquid crystal retarder is integrated into the illumination path of a DIC microscope for independent phase-shifting of sheared beams.

Main Results:

GROM successfully performed QPI on diverse specimens, including microbes, red blood cells, and optically thick plant roots (up to 300 μm).
The method functions without the need for sample fixation or clearing.
GROM simplifies configurations, reduces costs, and avoids energy losses in parallel imaging modalities.

Conclusions:

GROM expands the applicability of QPI to challenging, optically thick biological samples.
This technique offers a cost-effective and simplified approach to advanced bioimaging.
GROM enhances the study of cellular metabolism and structure in complex biological tissues.