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Updated: May 16, 2026

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

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Published on: April 7, 2014

Dynamic quantitative phase imaging for biological objects using a pixelated phase mask.

Katherine Creath¹, Goldie Goldstein

¹4D Technology Corp, 3280 E Hemisphere Loop, Ste 146, Tucson, AZ 85706, USA ; College of Optical Sciences, The University of Arizona, 1680 E University Blvd, Tucson, AZ 85721, USA ; Optineering, 2247 E La Mirada St, Tucson, AZ 85719, USA.

Biomedical Optics Express

|November 20, 2012

Summary

This study introduces a label-free dynamic quantitative phase imaging microscope for tracking live cell motions and changes. It enables instantaneous measurements of cell dynamics and morphology without needing contrast agents.

Keywords:

(110.3175) Interferometric imaging (120.5050) Phase measurement (170.3890) Medical optics instrumentation (170.6900) Three-dimensional microscopy (180.0180) Microscopy (180.3170) Interference microscopy

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Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Live cell imaging often requires labels or contrast agents, which can interfere with biological processes.
Tracking dynamic motions and morphological changes in cells is crucial for understanding cellular functions.
Existing microscopy techniques may lack the speed or specificity for real-time analysis of cellular dynamics.

Purpose of the Study:

To develop a dynamic quantitative phase imaging microscope for label-free, instantaneous measurement of cellular dynamics.
To enable tracking of dynamic motions and morphological changes within and among live cells.
To provide a method for quantifying changes in cell position and volume over time.

Main Methods:

Utilizing a pixelated phase mask for simultaneous measurement of multiple interference patterns.
Leveraging the polarization properties of light to track dynamic motions.
Obtaining optical path difference (OPD) and optical thickness (OT) data from phase images.
Implementing two processing routines to remove background surface shape for accurate quantification.

Main Results:

Demonstrated instantaneous measurements of dynamic motions in live cells without labels.
Successfully tracked dynamic motions and morphological changes using polarization-based interference patterns.
Quantified changes in cell position and volume over time after background correction.
Presented data from various moving biological organisms and cell cultures.

Conclusions:

The developed dynamic quantitative phase imaging microscope offers a powerful tool for label-free, real-time analysis of cellular dynamics.
This technique allows for non-invasive monitoring of cellular movements and structural alterations.
The method is applicable to a range of biological samples, including live cells and organisms.