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High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

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Single-shot quantitative phase-fluorescence imaging using cross-grating wavefront microscopy.

Baptiste Marthy¹, Maëlle Bénéfice¹, Guillaume Baffou²

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

Scientific Reports

|January 25, 2024

Summary

This summary is machine-generated.

This study presents a novel optical microscopy method for simultaneous fluorescence and phase imaging using a single camera and a diffraction grating. This technique eliminates image delay and registration issues for biological samples.

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Area of Science:

Optical microscopy
Biophotonics
Image processing

Background:

Simultaneous acquisition of fluorescence and phase images is challenging.
Existing methods often suffer from time delays or require image registration.
Advanced imaging techniques are crucial for detailed biological sample analysis.

Purpose of the Study:

To develop a new optical microscopy technique for simultaneous quantitative fluorescence and phase imaging.
To eliminate the delay and registration issues associated with separate image acquisition.
To demonstrate the technique's applicability on diverse biological samples.

Main Methods:

Utilized a 2-dimensional diffraction grating (cross-grating) with a 2-color camera.
Acquired fluorescence and wavefront images simultaneously from different color channels.
Employed image demodulation to retrieve quantitative phase and fluorescence information.

Main Results:

Successfully demonstrated simultaneous acquisition of fluorescence and phase images.
Achieved accurate image retrieval without time delay or registration.
Validated the method on fluorescent micro-beads, bacteria, and mammalian cells.

Conclusions:

The presented optical microscopy technique offers a robust solution for simultaneous fluorescence and phase imaging.
This method simplifies image acquisition and enhances the analysis of biological specimens.
The technique shows significant potential for various applications in life sciences research.