Accurate evaluation of size and refractive index for spherical objects in quantitative phase imaging
Optics Express
|May 3, 2018
Summary
This study introduces a new algorithm for quantitative phase imaging (QPI) to accurately measure the refractive index (RI) of spherical objects like cells, overcoming limitations of previous methods.
Area of Science:
- Optical physics
- Biophysics
- Microscopy
Background:
- Quantitative phase imaging (QPI) is crucial for label-free cell analysis.
- Accurate measurement of refractive index (RI) and size of spherical objects in QPI is challenging.
- Existing methods often neglect light-scattering effects, limiting RI accuracy.
Purpose of the Study:
- To develop and validate a novel QPI fitting algorithm for precise RI and size decoupling.
- To improve the accuracy of RI measurements for spherical objects in QPI.
- To provide practical guidelines for analyzing spherical objects using QPI.
Main Methods:
- Development of a novel QPI fitting algorithm.
- Integration of Mie theory and a semi-analytical, corrected Rytov approach.
- In silico and experimental validation using diverse spherical objects and noise conditions.
Main Results:
- The algorithm reliably uncouples RI and size from QPI data.
- Demonstrated validity across various objects including cells, droplets, and beads.
- Performance assessed under different noise conditions.
Conclusions:
- The novel algorithm offers a robust solution for accurate RI determination in QPI.
- The approach overcomes limitations of geometrical methods by incorporating light-scattering.
- Provides practical insights for enhanced spherical object analysis in QPI.
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