Sub-microscopic analysis of t-tubule geometry in living cardiac ventricular myocytes using a shape-based analysis method

Autor: Eva A. Rog-Zielinska, Peter Kohl, Cherrie H.T. Kong, Clive H. Orchard, Mark B. Cannell
Rok vydání: 2017
Předmět:
0301 basic medicine
Heart Ventricles
Bristol Heart Institute
Geometry
Image processing
030204 cardiovascular system & hematology
Fluorescent dyes
03 medical and health sciences
chemistry.chemical_compound
Mice
SS
surface sarcolemma
0302 clinical medicine
Cardiac myocyte
FWHM
full-width at half maximum
Image Processing
Computer-Assisted
Animals
Myocytes
Cardiac
V:SA
volume to surface area ratio
Molecular Biology
EM
electron microscopy
Microscopy
Confocal
Chemistry
Confocal imaging
E-C
excitation-contraction
Coupling (electronics)
Calcein
Full width at half maximum
Sarcoplasmic Reticulum
030104 developmental biology
Tubule
Electron tomography
Volume (thermodynamics)
Surface-area-to-volume ratio
IAS
T-tubules
TTs
transverse-axial tubules
Original Article
Rabbits
Cardiology and Cardiovascular Medicine
ET
electron microscopic tomography
DTT
t-tubule diameter
Zdroj: Journal of Molecular and Cellular Cardiology
Kong, C H T, Rog-Zielinska, E A, Kohl, P, Orchard, C & Cannell, M B 2017, ' Sub-microscopic analysis of t-tubule geometry in living cardiac ventricular myocytes using a shape-based analysis method ', Journal of Molecular and Cellular Cardiology, vol. 108, pp. 1-7 . https://doi.org/10.1016/j.yjmcc.2017.05.003
ISSN: 1095-8584
Popis: Transverse-axial tubules (TTs) are key structures involved in cardiac excitation-contraction coupling and can become deranged in disease. Although optical measurement of TTs is frequently employed to assess TT abundance and regularity, TT dimensions are generally below the diffraction limit of optical microscopy so determination of tubule size is problematic. TT diameter was measured by labeling both local surface membrane area and volume with fluorescent probes (FM4-64 and calcein, respectively), correcting image asymmetry by image processing and using the relationship between surface area and volume for a geometric primitive. This method shows that TTs have a mean (± SEM) diameter of 356 ± 18 nm in rabbit and 169 ± 15 nm in mouse (p
Highlights • Living cardiomyocytes were dual-labeled with fluorescent surface and volume probes. • A novel 3D image processing strategy enabled calculation of t-tubule diameter. • The method shows rabbit and mouse t-tubules have quite different morphologies. • Mean diameters of rabbit and mouse t-tubules were 360 and 170 nm, respectively. • Estimated electrical space constants are sufficient to ensure electrical uniformity.
Databáze: OpenAIRE
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