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Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...

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Related Experiment Video

Updated: Apr 28, 2026

Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment

Whole-cell Super-Resolution Imaging via DNA-PAINT on a Spinning Disk Confocal with Optical Photon Reassignment

Published on: January 6, 2026

Spectroscopic DNA-PAINT for simultaneous multiplexed super-resolution microscopy.

Md Abul Shahid¹, Kripa Patel¹, David A Miller²

¹Molecular Analytics and Photonics (MAP) Lab, Department of Textile Engineering, Chemistry and Science, North Carolina State University, Raleigh, NC 27606.

Biorxiv : the Preprint Server for Biology

|April 27, 2026

Summary

This summary is machine-generated.

Spectroscopic DNA-PAINT (sDNA-PAINT) enables simultaneous super-resolution imaging of multiple targets with high precision. This advanced technique improves spectral fidelity and photon budgets for complex biological samples.

Keywords:

DNA-PAINT multiplexing single-molecule spectroscopy super-resolution microscopy

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Last Updated: Apr 28, 2026

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Area of Science:

Biophysics
Optical Microscopy
Nanotechnology

Background:

Simultaneous multiplexed super-resolution imaging is crucial for understanding molecular interactions but faces challenges with spectral overlap and precision.
Current single-molecule localization microscopy (SMLM) methods have limitations in distinguishing multiple fluorophores simultaneously.

Purpose of the Study:

To introduce and validate spectroscopic DNA-PAINT (sDNA-PAINT) for high-fidelity, simultaneous multiplexed super-resolution imaging.
To enhance spectral precision and photon budgets in SMLM for improved multiplexing capabilities.

Main Methods:

Integration of DNA-PAINT with spectroscopic SMLM (sSMLM) to create the sDNA-PAINT framework.
Utilized DNA Origami Nanorulers for *in vitro* and fixed-cell imaging experiments.
Tested sDNA-PAINT with various fluorophore families (Rhodamine, Cyanine, Oxazine) and compared performance against conventional SMLM conditions.

Main Results:

sDNA-PAINT significantly improved spectral precision and photon budgets compared to conventional methods.
Demonstrated reliable single-molecule discrimination even for spectrally overlapping dyes.
Achieved accurate spatial reconstruction and high classification accuracy in dual-target cellular imaging.

Conclusions:

sDNA-PAINT enables high-fidelity simultaneous multiplexed super-resolution imaging within a single acquisition.
This framework offers a pathway toward high-throughput simultaneous multiplexed super-resolution interaction imaging.
sDNA-PAINT overcomes spectral limitations, advancing the field of super-resolution microscopy.