1 July 2009 Three-dimensional cardiac architecture determined by two-photon microtomy
Author Affiliations +
J. of Biomedical Optics, 14(4), 044029 (2009). doi:10.1117/1.3200939
Cardiac architecture is inherently three-dimensional, yet most characterizations rely on two-dimensional histological slices or dissociated cells, which remove the native geometry of the heart. We previously developed a method for labeling intact heart sections without dissociation and imaging large volumes while preserving their three-dimensional structure. We further refine this method to permit quantitative analysis of imaged sections. After data acquisition, these sections are assembled using image-processing tools, and qualitative and quantitative information is extracted. By examining the reconstructed cardiac blocks, one can observe end-to-end adjacent cardiac myocytes (cardiac strands) changing cross-sectional geometries, merging and separating from other strands. Quantitatively, representative cross-sectional areas typically used for determining hypertrophy omit the three-dimensional component; we show that taking orientation into account can significantly alter the analysis. Using fast-Fourier transform analysis, we analyze the gross organization of cardiac strands in three dimensions. By characterizing cardiac structure in three dimensions, we are able to determine that the α crystallin mutation leads to hypertrophy with cross-sectional area increases, but not necessarily via changes in fiber orientation distribution.
Hayden Huang, Catherine MacGillivray, Hyuk-Sang Kwon, Jan Lammerding, Jeffrey Robbins, Richard T. Lee, Peter T. C. So, "Three-dimensional cardiac architecture determined by two-photon microtomy," Journal of Biomedical Optics 14(4), 044029 (1 July 2009). https://doi.org/10.1117/1.3200939




3D image processing


Two photon excitation microscopy

Data acquisition

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