Nanoscopy on-a-chip: super-resolution imaging on the millimeter scale.

Øystein I Helle,Balpreet S Ahluwalia,David A Coucheron,Jean-Claude Tinguely,Cristina I Øie

doi:10.1364/oe.27.006700

Abstract

Optical nanoscopy techniques can image intracellular structures with high specificity at sub-diffraction limited resolution, bridging the resolution gap between optical microscopy and electron microscopy. So far conventional nanoscopy lacks the ability to generate high throughput data, as the imaged region is small. Photonic chip-based nanoscopy has demonstrated the potential for imaging large areas, but at a lateral resolution of 130 nm. However, all the existing super-resolution methods provide a resolution of 100 nm or better. In this work, chip-based nanoscopy is demonstrated with a resolution of 75 nm over an extraordinarily large area of 0.5 mm × 0.5 mm, using a low magnification and high N.A. objective lens. Furthermore, the performance of chip-based nanoscopy is benchmarked by studying the localization precision and illumination homogeneity for different waveguide widths. The advent of large field-of-view chip-based nanoscopy opens up new routes in diagnostics where high throughput is needed for the detection of non-diffuse disease, or rare events such as the early detection of cancer.

Highlights

For a long time, the spatial resolution in optical microscopy was believed to be bound by the diffraction limit
From the early 90s up until now a range of techniques developed that aim to produce images with spatial resolution way beyond that of the diffraction limit. This field is commonly known as super-resolution optical microscopy or optical nanoscopy, and has given biologists tools to observe living intracellular structures with unprecedented high resolution
For conventional immunolabeling at room temperature using antibody-binding of fluorophores, this typically means a few tens of nanometers

Summary

Introduction

The spatial resolution in optical microscopy was believed to be bound by the diffraction limit. From the early 90s up until now a range of techniques developed that aim to produce images with spatial resolution way beyond that of the diffraction limit. This field is commonly known as super-resolution optical microscopy or optical nanoscopy, and has given biologists tools to observe living intracellular structures with unprecedented high resolution. Improvements in the nanoscopy methods have pushed the spatial resolution towards the ultimate limit i.e. the physical size of the fluorescent labels [ 2]. For conventional immunolabeling at room temperature using antibody-binding of fluorophores, this typically means a few tens of nanometers

Methods

Results

Conclusion