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High speed sCMOS-based oblique plane microscopy applied to the study of calcium dynamics in cardiac myocytes

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Title: High speed sCMOS-based oblique plane microscopy applied to the study of calcium dynamics in cardiac myocytes
Authors: Sikkel, MB
Kumar, S
Maioli, V
Rowlands, C
Gordon, F
Harding, SE
Lyon, A
MacLeod, KT
Dunsby, CW
Item Type: Journal Article
Abstract: blique plane microscopy (OPM) is a form of light sheet microscopy that uses a single high numerical aperture microscope objective for both fluorescence excitation and collection. In this paper, measurements of the relative collection efficiency of OPM are presented. An OPM system incorporating two sCMOS cameras is then introduced that enables single isolated cardiac myocytes to be studied continuously for 22 seconds in two dimensions at 667 frames per second with 960 × 200 pixels and for 30 seconds with 960 × 200 × 20 voxels at 25 volumes per second. In both cases OPM is able to record in two spectral channels, enabling intracellular calcium to be studied via the probe Fluo-4 AM simultaneously with the sarcolemma and transverse tubule network via the membrane dye Cellmask Orange. The OPM system was then applied to determine the spatial origin of spontaneous calcium waves for the first time and to measure the cell transverse tubule structure at their point of origin. Further results are presented to demonstrate that the OPM system can also be used to study calcium spark parameters depending on their relationship to the transverse tubule structure.
Issue Date: 1-Mar-2016
Date of Acceptance: 16-Sep-2015
URI: http://hdl.handle.net/10044/1/26357
DOI: 10.1002/jbio.201500193
ISSN: 1864-0648
Publisher: Wiley-VCH Verlag
Start Page: 311
End Page: 323
Journal / Book Title: Journal of Biophotonics
Volume: 9
Issue: 3
Copyright Statement: © 2015 The Author. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and repro-duction in any medium, provided the original work is properly cited.
Sponsor/Funder: Engineering & Physical Science Research Council (EPSRC)
The Royal Society
British Heart Foundation
British Heart Foundation
Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
British Heart Foundation
British Heart Foundation
British Heart Foundation
Funder's Grant Number: EP/H03238X/1
N/A
RG/12/18/30088
RE/08/002/23906
092852/Z/10/Z
BB/I023801/1
FS/11/67/28954
PG/09/032/27241
SP/16/2/32004
Keywords: Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemical Research Methods
Biophysics
Optics
Biochemistry & Molecular Biology
high-speed imaging
light-sheet microscopy
calcium dynamics
3D imaging
Oblique Plane Microscopy
SARCOPLASMIC-RETICULUM CALCIUM
CA2+ SPARKS
FLUORESCENCE MICROSCOPY
REDUCED SYNCHRONY
T-TUBULES
RELEASE
WAVES
3D imaging
Oblique Plane Microscopy
calcium dynamics
high-speed imaging
light-sheet microscopy
Animals
Calcium Signaling
Imaging, Three-Dimensional
Metals
Microscopy
Myocytes, Cardiac
Optical Phenomena
Oxides
Rats
Semiconductors
Time Factors
Myocytes, Cardiac
Animals
Rats
Oxides
Metals
Imaging, Three-Dimensional
Microscopy
Calcium Signaling
Semiconductors
Time Factors
Optical Phenomena
Optoelectronics & Photonics
0205 Optical Physics
0304 Medicinal and Biomolecular Chemistry
1004 Medical Biotechnology
Publication Status: Published
Online Publication Date: 2015-10-21
Appears in Collections:Physics
Photonics
National Heart and Lung Institute
Faculty of Medicine
Faculty of Natural Sciences