Double-exposure optical sectioning structured illumination microscopy based on Hilbert transform reconstruction

Xing Zhou; Ming Lei; Dan Dan; Baoli Yao; Jia Qian; Shaohui Yan; Yanlong Yang; Junwei Min; Tong Peng; Tong Ye; Guangde Chen

doi:10.1371/journal.pone.0120892

Double-exposure optical sectioning structured illumination microscopy based on Hilbert transform reconstruction

PLoS One. 2015 Mar 23;10(3):e0120892. doi: 10.1371/journal.pone.0120892. eCollection 2015.

Authors

Xing Zhou¹, Ming Lei², Dan Dan², Baoli Yao², Jia Qian², Shaohui Yan², Yanlong Yang², Junwei Min², Tong Peng², Tong Ye³, Guangde Chen⁴

Affiliations

¹ State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics, Chinese Academy of Sciences, Xi'an, 710119, China; School of Science, Xi'an Jiaotong University, Xi'an, 710049, China.
² State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics, Chinese Academy of Sciences, Xi'an, 710119, China.
³ Department of Bioengineering, Clemson University, Clemson-MUSC Bioengineering Program, Charleston, South Carolina, 29425, United States of America.
⁴ School of Science, Xi'an Jiaotong University, Xi'an, 710049, China.

Abstract

Structured illumination microscopy (SIM) with axially optical sectioning capability has found widespread applications in three-dimensional live cell imaging in recent years, since it combines high sensitivity, short image acquisition time, and high spatial resolution. To obtain one sectioned slice, three raw images with a fixed phase-shift, normally 2π/3, are generally required. In this paper, we report a data processing algorithm based on the one-dimensional Hilbert transform, which needs only two raw images with arbitrary phase-shift for each single slice. The proposed algorithm is different from the previous two-dimensional Hilbert spiral transform algorithm in theory. The presented algorithm has the advantages of simpler data processing procedure, faster computation speed and better reconstructed image quality. The validity of the scheme is verified by imaging biological samples in our developed DMD-based LED-illumination SIM system.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Algorithms
Image Processing, Computer-Assisted / methods*
Imaging, Three-Dimensional / methods*
Microscopy / methods*

Grants and funding

This research is supported by the Natural Science Foundation of China (NSFC) under Grant No. 61275193, 61275191, 61377008, 11404389 and the National Basic Research Program (973 Program) of China under grant No. 2012CB921900. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.