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Label-free high-resolution white light quantitative phase nanoscopy system.

Shilpa Tayal¹, Shubham Tiwari¹, Dalip Singh Mehta¹

¹Bio-photonics and Green Photonics Laboratory, Department of Physics, Indian Institute of Technology Delhi, New Delhi, India.

Journal of Biophotonics

|January 5, 2023

Summary

We developed a high-resolution white light quantitative phase nanoscopy (WLQPN) system. This advanced imaging technique visualizes nanoparticles and subcellular structures with improved resolution for biological specimens.

Keywords:

deconvolution high-resolution imaging phase nanoscopy phase-shifting white light

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Area of Science:

Optical microscopy
Nanotechnology
Biophysics

Background:

Traditional phase imaging methods often face limitations in achieving super-resolution.
Fourier analysis methods in phase imaging can restrict resolution below the diffraction limit.
Visualizing nanoscale biological structures requires advanced imaging techniques.

Purpose of the Study:

To present a high-resolution white light quantitative phase nanoscopy (WLQPN) system.
To achieve super-resolution in phase imaging for biological specimens.
To overcome the resolution limitations of conventional Fourier analysis methods.

Main Methods:

Utilized a five-phase shifting technique for initial phase map recovery, preserving full detector resolution.
Employed Richardson-Lucy total variation deconvolution on the complex field for super-resolution phase imaging.
Tested the system on a USAF resolution chart, polystyrene beads, and Escherichia coli bacteria.

Main Results:

The WLQPN system successfully visualized 200 nm polystyrene beads.
Subcellular features of Escherichia coli bacteria were clearly observed.
Demonstrated significant resolution improvement compared to standard methods.

Conclusions:

The developed WLQPN system provides high-resolution phase imaging capabilities.
The combination of phase shifting and deconvolution achieves super-resolution for nanoscale biological imaging.
This technique offers a valuable tool for visualizing nanoparticles and subcellular details.