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Homographically generated light sheets for the microscopy of large specimens.

Published version
Peer-reviewed

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Authors

Russell, Craig T 
Rees, Eric J 
Kaminski, Clemens F 

Abstract

We compare the performance of linear and nonlinear methods for aligning the excitation and detection planes throughout volumes of large specimens in digitally scanned light sheet microscopy. An effective nonlinear method involves the registration of four corner extrema of the imaging volume via a projective transform. We show that this improves the light collection efficiency of the commonly used three-point affine registration by an average of 42% over a typical specimen volume. Accurate illumination/detection registration methods are now pertinent to biological research in view of current trends towards imaging large or expanded samples, at depth, with diffraction limited resolution.

Description

Keywords

Fluorescent Dyes, Imaging, Three-Dimensional, Light, Microscopy

Journal Title

Opt Lett

Conference Name

Journal ISSN

0146-9592
1539-4794

Volume Title

43

Publisher

The Optical Society
Sponsorship
EPSRC (1362132)
Engineering and Physical Sciences Research Council (EP/H018301/1)
Medical Research Council (MR/K02292X/1)
Medical Research Council (MR/K015850/1)
Wellcome Trust (089703/Z/09/Z)
Biotechnology and Biological Sciences Research Council (BB/H023917/1)
Medical Research Council (G0902243)
Engineering and Physical Sciences Research Council (EP/L015889/1)