Method/Kit:
Confocal Microscopy

Measurement Type Imaging
Description Leica SP5 Inverted Confocal Microscope.
Excitation frequencies used: 405, 488, 633

Target/Analytes with this Method/Kit
Target/Analyte Short Name Description
Cross Sectional Area CSA Cross-Sectional Area of myobundles as observed by image staining.
Cx43+ area Cx43+ Quantification of percent imaged area positive for Cx43
Fiber diameter FD Fiber diameter of individual myofibers in the engineered myobundles
Fraction Nuclei in SAA+ Fibers In SAA/ Total Nuclei Number of nuclei in SAA+ fibers in engineered myobundles observed by immunofluorescence.
Myofiber Length Fiber Length Length of myofibers in engineered myobundles observed by immunofluorescence.
Myofiber Orientation Fiber Alignment Fiber alignment in engineered myobundles observed by immunofluorescence.
MyoG+ Nuclei MyoG Fraction MyoG+ nuclei in myobundles as observed by image staining.
Nuclear Circularity Nuc Circ Characteristics of individual DAPI-stained nuclei, including circularity and lengths of major and minor nuclear axes, were determined using particle analysis in ImageJ-Fiji, in a 0.006 mm2 area in each section.
Nuclear Dispersion ND The Nuclear dispersion was determined in DAPI-stained whole-mount sections by analysis of 0.3 mm2 areas using the Directionality plug-in ImageJ-Fiji by randomly analyzing 0.003 mm2 areas of each tissue after software auto-thresholding with the default method.
Nuclear Elongation NE characteristics of individual DAPI-stained nuclei, including circularity and lengths of major and minor nuclear axes, were determined using particle analysis in ImageJ-Fiji, in a 0.006 mm2 area in each section.
Nuclei Count/ Cross-Sectional Area Nuclei Count Nuclei / Cross-Sectional Area of myobundles as observed by image staining.
Percent striated fibers Striated fibers Number of striated fibers in engineered myobundles observed by immunofluorescence.
SAA+ Area/ Cross-Sectional Area SAA+ SAA+ Area/ Cross-Sectional Area of myobundles as observed by image staining.
SAA+ Area in LS SAA+ Area SAA+ Area in engineered myobundles observed by immunofluorescence.
Sarcomere Dispersion SD The direction and dispersion of sarcomeres in whole-mount sarcomeric α-actinin (SAA) stained cardiobundles were determined using the Directionality plug-in ImageJ-Fiji by randomly analyzing 0.003 mm2 areas of each tissue after software auto-thresholding with the default method
TUNEL+ Nuclei TUNEL+ quantifications of fractions of TUNEL+ nuclei
Cell Morphology Cell Morphology Imaging, typically with transmitted light, to see the cell morphology.

Studies Utilizing this Method/Kit
Study Start Date Study Types Description
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