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Updated: Jul 6, 2025

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

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

Published on: November 16, 2019

Phase diversity-based wavefront sensing for fluorescence microscopy.

Courtney Johnson¹, Min Guo^2,3, Magdalena C Schneider¹

¹Janelia Research Campus, Howard Hughes Medical Institute (HHMI), Ashburn, VA, USA.

Biorxiv : the Preprint Server for Biology

|January 3, 2024

Summary

This summary is machine-generated.

We developed a rapid phase diversity method for adaptive optics (AO) in fluorescence microscopy. This technique quickly corrects optical aberrations, significantly improving image quality for biological samples.

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Area of Science:

Microscopy
Biophysics
Optical Engineering

Background:

Fluorescence microscopy is crucial for biological research but limited by optical aberrations.
Aberrations degrade image contrast, resolution, and signal, reducing data quality.
Existing adaptive optics (AO) methods are often too complex or slow for widespread use.

Approach:

Introduced a rapid, sensitive, and robust wavefront sensing scheme based on phase diversity, adapted from astronomy.
Achieved accurate wavefront sensing with < λ/35 RMS error using minimal measurements.
Integrated AO correction without requiring additional hardware beyond a deformable mirror.

Key Points:

Demonstrated >100-fold faster deformable mirror calibration (~100 ms scale) compared to existing methods.
Successfully sensed and corrected severe optical aberrations (RMS wavefront distortion > λ/2).
Restored diffraction-limited imaging performance on extended biological samples.

Conclusions:

The phase diversity method offers a fast, sensitive, and robust solution for AO in microscopy.
This approach overcomes limitations of current AO techniques, making advanced optical correction more accessible.
Enables high-quality imaging of biological specimens previously hindered by optical aberrations.