Super-resolution fluorescence microscopy by stepwise optical saturation

Article

Abstract

Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the super-resolution microscopy is not feasible in many applications. In this paper, we propose and demonstrate a saturation-based super-resolution fluorescence microscopy technique that can be easily implemented and requires neither additional hardware nor complex post-processing. The method is based on the principle of stepwise optical saturation (SOS), where M steps of raw fluorescence images are linearly combined to generate an image with a 𝑀−−√-fold increase in resolution compared with conventional diffraction-limited images. For example, linearly combining (scaling and subtracting) two images obtained at regular powers extends the resolution by a factor of 1.4 beyond the diffraction limit. The resolution improvement in SOS microscopy is theoretically infinite but practically is limited by the signal-to-noise ratio. We perform simulations and experimentally demonstrate super-resolution microscopy with both one-photon (confocal) and multiphoton excitation fluorescence. We show that with the multiphoton modality, the SOS microscopy can provide super-resolution imaging deep in scattering samples.

Attributes

Attribute NameValues
Creator
  • Yide Zhang

  • Prakash Nallathamby

  • Genevieve D. Vigil

  • Aamir A. Khan

  • Devon E. Mason

  • Joel D. Boerckel

  • Ryan K. Roeder

  • Scott S. Howard

Journal or Work Title
  • Biomedical Optics Express

Volume
  • 9

Issue
  • 4

First Page
  • 1613

Last Page
  • 1619

Publication Date
  • 2018-04

Publisher
  • OSA - The Optical Society

Date Created
  • 2018-10-31

Language
  • English

Departments and Units
Record Visibility and Access Public
Content License
  • All rights reserved

Digital Object Identifier

doi:10.1364/BOE.9.001613

This DOI is the best way to cite this article.