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Updated: Jun 10, 2025

Quantitative Optical Microscopy: Measurement of Cellular Biophysical Features with a Standard Optical Microscope
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Quantitative phase microscopies: accuracy comparison.

Patrick C Chaumet1, Pierre Bon2, Guillaume Maire1

  • 1Institut Fresnel, CNRS, Aix Marseille Univ, Centrale Med, Marseille, France.

Light, Science & Applications
|October 11, 2024
PubMed
Summary
This summary is machine-generated.

Quantitative phase microscopies (QPMs) offer label-free bio-imaging insights. This review compares 8 QPM techniques, detailing their precision and accuracy, aiding selection for specific applications.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Quantitative Imaging

Background:

  • Quantitative phase microscopies (QPMs) are crucial for bio-imaging, providing label-free data on mass distribution and transport.
  • QPMs complement fluorescence imaging by avoiding photobleaching and phototoxicity.
  • Selecting the optimal QPM technique for specific applications can be challenging due to the variety available.

Purpose of the Study:

  • To provide a comprehensive tutorial review comparing the main QPM techniques.
  • To focus on the accuracy of QPMs, specifically measurement precision and trueness.
  • To assist researchers in choosing the most suitable QPM for their needs.

Main Methods:

  • Comparison of 8 QPM techniques: digital holographic microscopy (DHM), cross-grating wavefront microscopy (CGM), diffraction phase microscopy (DPM), differential phase-contrast (DPC) microscopy, phase-shifting interferometry (PSI) imaging, Fourier phase microscopy (FPM), spatial light interference microscopy (SLIM), and transport-of-intensity equation (TIE) imaging.
  • Utilized a custom numerical toolbox based on discrete dipole approximation (IF-DDA) to model electromagnetic fields.
  • Upgraded the toolbox to simulate various QPMs and incorporate shot noise effects.

Main Results:

  • Digital holographic microscopy (DHM) and phase-shifting interferometry (PSI) are largely artifact-free, primarily affected by coherent noise.
  • Cross-grating wavefront microscopy (CGM), differential phase-contrast (DPC) microscopy, diffraction phase microscopy (DPM), and transport-of-intensity equation (TIE) imaging show a precision-trueness trade-off adjustable via experimental parameters.
  • Fourier phase microscopy (FPM) and spatial light interference microscopy (SLIM) exhibit inherent artifacts, limiting their quantitative accuracy, especially for large biological specimens like eukaryotic cells.

Conclusions:

  • The study provides a quantitative comparison of major QPM techniques based on precision and trueness.
  • Results guide the selection of QPMs by highlighting their strengths and limitations regarding accuracy and artifacts.
  • The developed numerical toolbox facilitates the modeling and analysis of QPM performance.