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Updated: Nov 26, 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

Practical application of microsphere samples for benchmarking a quantitative phase imaging system.

Edward Kwee¹, Alexander Peterson¹, Michael Halter¹

¹Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, Maryland, USA.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|December 11, 2020

Summary

This summary is machine-generated.

Quantitative phase imaging (QPI) offers instrument-independent cell dry mass monitoring. This study introduces microsphere-based reference materials to calibrate QPI systems, ensuring accurate optical pathlength measurements for biological samples.

Keywords:

HEK293 microspheres quantitative phase imaging reference materials

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Last Updated: Nov 26, 2025

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

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Published on: April 7, 2014

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Published on: April 26, 2014

Area of Science:

Biophysics
Optical Metrology

Background:

Quantitative phase imaging (QPI) measures cell dry mass via optical pathlength.
QPI measurements are theoretically instrument-independent.
Standardized reference materials are needed to ensure QPI accuracy across different instruments.

Purpose of the Study:

To evaluate microsphere-based reference materials for QPI system benchmarking.
To establish calibration standards for accurate optical pathlength measurements.
To validate QPI for precise dry mass quantification.

Main Methods:

Investigated seven combinations of microspheres in refractive index-matching media.
Utilized poly(methyl methacrylate) microspheres and mineral oil.
Assessed illumination apertures, signal-to-noise ratios, and focus positions for accurate measurements.

Main Results:

Developed microsphere/media reference materials for QPI benchmarking.
Verified instrument settings for accurate optical pathlength measurements.
Applied the reference material to benchmark QPI dry mass measurements of HEK293 cells.

Conclusions:

Microsphere-based reference materials are effective for QPI system calibration.
This approach ensures the accuracy and reliability of QPI dry mass measurements.
Validated QPI performance for biological cell analysis.