Work

Fast, super resolution imaging via Bessel-beam stimulated emission depletion microscopy

Year: 2014

Type: article

Abstract: A substantial advantage of stimulated emission depletion (STED) microscopy over other super-resolution methods is that images can be acquired in real-time without any post-processing. However imaging speed and photodamage are two major concerns for STED imaging of whole cells. Here we propose a new microscopy method we have termed Bessel-Beam STED (or BB-STED) that overcomes both of these limitations of conventional STED microscopy. In the proposed method, rather than exciting a single STED spot in the sample, an entire line of the sample is illuminated. This line-scanning technique dramatically increases the speed of STED. In addition, plane-illumination by scanning of the line across the focal plane of a detection objective limits the light to a thin layer of the sample and thus significantly reduces photobleaching and photodamage above and below the focal plane compared to epi-illumination. Using the organic dye Atto647N as an example, we calculated the STED power required to break the diffraction limit. The results presented here will be used to guide future experimental designs.

Source: Optics Express

DOI: 10.1364/oe.22.012398

Authors: P. Zhang, P. M. Goodwin, J. H. Werner

Institutions: Los Alamos National Laboratory, Center for Integrated Nanotechnologies

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,270

Funders: Basic Energy Sciences, Laboratory Directed Research and Development, Los Alamos National Laboratory