Rapid 3D image scanning microscopy with multi-spot excitation and double-helix point spread function detection

Refocusing after Scanning using Helical phase engineering (RESCH) microscopy has previously been demonstrated to provide volumetric information from a single 2D scan. However, the practical application of this method is challenging due to its limited image acquisition speed and spatial resolution. H...

Full description

Saved in:

Bibliographic Details
Published in:	Optics express Vol. 26; no. 18; p. 23585
Main Authors:	Li, Siwei, Wu, Jingjing, Li, Heng, Lin, Danying, Yu, Bin, Qu, Junle
Format:	Journal Article
Language:	English
Published:	United States 03.09.2018
ISSN:	1094-4087, 1094-4087
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Refocusing after Scanning using Helical phase engineering (RESCH) microscopy has previously been demonstrated to provide volumetric information from a single 2D scan. However, the practical application of this method is challenging due to its limited image acquisition speed and spatial resolution. Here, we report on a combination of RESCH and multifocal structured illumination microscopy (MSIM) to improve the image acquisition speed and spatial resolution. A phase mask is introduced to modulate the conventional point spread function (PSF) to the double-helix PSF (DH-PSF), which provides volumetric information, and meanwhile, sparse multifocal illumination patterns are generated by a digital micromirror device (DMD) for parallel 3D subdiffractive imaging information acquisition. We also present a strategy for processing the collected raw data with a Richardson-Lucy deconvolution and pixel reassignment algorithm to improve the spatial resolution of the depth estimation and imaging performance. The proposed 3D image scanning microscopy can record 3D specimen information and the corresponding depth information from a single multi-spot 2D planar scan, which ensures faster data acquisition, larger field of view, and higher spatial resolution than RESCH. Finally, we demonstrate the capability of our system with actual experiments.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1094-4087 1094-4087
DOI:	10.1364/OE.26.023585