Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/52517
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Type: Journal article
Title: Second-harmonic generation and two-photon-excited autofluorescence microscopy of cardiomyocytes: quantification of cell volume and myosin filaments
Author: Wallace, S.
Morrison, J.
Botting, K.
Kee, T.
Citation: Journal of Biomedical Optics, 2008; 13(6):1
Publisher: SPIE-Int Society Optical Engineering
Issue Date: 2008
ISSN: 1083-3668
1560-2281
Statement of
Responsibility: 
Samuel J. Wallace, Janna L. Morrison, Kimberley J. Botting and Tak W. Kee
Abstract: The ability to quantify changes in cardiomyocyte and myosin volume across gestation and in response to intrauterine insults will lead to a better understanding of the link between low birth weight and an increased risk of heart disease in adult life. We present the use of second-harmonic generation (SHG) and two-photon excitation autofluorescence (TPEF) microscopy to image unstained isolated fetal cardiomyocytes. The simultaneous collection of these two images provides a wealth of information on the morphology of cardiomyocytes. The SHG signal provides high-contrast images of myosin filaments and the TPEF signal can be used to clearly visualize cell morphology. A potential issue may arise if SHG microscopy is performed exclusively due to the lack of sensitivity to distinguish between mononucleated and binucleated cardiomyocytes. However, TPEF microscopy has the ability to efficiently separate the two types of cardiomyocytes. In addition, quantitative analysis of the SHG and TPEF images enables quantification of myosin filament level and accurate determination of cell volume. In short, we demonstrate that advanced nonlinear optical microscopy can be used to answer key physiological questions in the early origins of adult health with increased accuracy and speed compared to previously used methods.
Keywords: Fetal Heart; Cells, Cultured; Myocytes, Cardiac; Animals; Sheep; Myosins; Image Interpretation, Computer-Assisted; Microscopy, Fluorescence, Multiphoton; Cell Size; Algorithms; Artificial Intelligence; Pattern Recognition, Automated
Description: Copyright © 2008 Society of Photo-Optical Instrumentation Engineers
RMID: 0020084359
DOI: 10.1117/1.3027970
Grant ID: http://purl.org/au-research/grants/nhmrc/511341
Appears in Collections:Chemistry publications
Environment Institute publications

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