Open AccessDeconvolution approach for 3D scanning microscopy with helical phase engineering
Author(s) -
Clemens Roider,
Rainer Heintzmann,
Rafael Piestun,
Alexander Jesacher
Publication year - 2016
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.24.015456
Subject(s) - deconvolution , microscopy , optics , point spread function , resolution (logic) , pixel , optical sectioning , laser scanning , image resolution , phase (matter) , light sheet fluorescence microscopy , materials science , image processing , microscope , digital holographic microscopy , confocal microscopy , scanning confocal electron microscopy , computer science , artificial intelligence , physics , laser , image (mathematics) , quantum mechanics
RESCH (refocusing after scanning using helical phase engineering) microscopy is a scanning technique using engineered point spread functions which provides volumetric information. We present a strategy for processing the collected raw data with a multi-view maximum likelihood deconvolution algorithm, which inherently comprises the resolution gain of pixel-reassignment microscopy. The method, which we term MD-RESCH (for multi-view deconvolved RESCH), achieves in our current implementation a 20% resolution advantage along all three axes compared to RESCH and confocal microscopy. Along the axial direction, the resolution is comparable to that of image scanning microscopy. However, because the method inherently reconstructs a volume from a single 2D scan, a significantly higher optical sectioning becomes directly visible to the user, which would otherwise require collecting multiple 2D scans taken at a series of axial positions. Further, we introduce the use of a single-helical detection PSF to obtain an increased post-acquisition refocusing range. We present data from numerical simulations as well as experiments to confirm the validity of our approach.