Isotropic 3D Super-resolution Imaging with a Self-bending Point Spread Function

Shu Jia,Xiaowei Zhuang,Joshua C Vaughan

doi:10.1364/fio.2013.ftu2f.2

Abstract

We introduce a self-bending point spread function (SB-PSF) based on Airy beams for three-dimensional (3D) super-resolution fluorescence imaging. We designed a side-lobe-free SB-PSF for the incoherent fluorescence emission and implemented a two-channel detection scheme for the SB-PSF to enable unambiguous 3D localization of fluorescent molecules. The lack of diffraction and the propagation-dependent lateral bending make the SB-PSF ideal for precise 3D localization of molecules over a large imaging depth. Using this SB-PSF, we demonstrated super-resolution imaging with isotropic localization precisions of 10-15 nm in all three dimensions over a 3 μm imaging depth without sample scanning.

Highlights

We report an approach to localize individual fluorophores with both isotropic 3D localization precision and substantial improvement in the imaging depth based on a self-bending point spread function (SB-PSF) derived from Airy beams[2, 3]
Airy beams can be generated based on the consideration that a 2D exponentially truncated Airy function Ai(x/a0, y/a0) is the Fourier transform of a Gaussian beam modulated by a cubic spatial phase, where (x, y) and are the spatial coordinates and corresponding wavevector components, respectively[2, 3]
Fluorescence emissions from individual molecules can in principle be converted into Airy beams if we introduce the cubic spatial phase with a spatial light modulator (SLM) placed at the Fourier plane in the detection path of the microscope (Figure 1a and Supplementary Figures 1 and 2)

Summary

Introduction

We report an approach to localize individual fluorophores with both isotropic 3D localization precision and substantial improvement in the imaging depth based on a SB-PSF derived from Airy beams[2, 3]. Fluorescence emissions from individual molecules can in principle be converted into Airy beams if we introduce the cubic spatial phase with a spatial light modulator (SLM) placed at the Fourier plane in the detection path of the microscope (Figure 1a and Supplementary Figures 1 and 2).

Results

Conclusion