Home

Browse
Communities
& Collections

Issue Date
Author
Title
Subject

Sign on to:

My Account
authorized users

Edit Profile

Help
 Please use this identifier to cite or link to this item: http://hdl.handle.net/1807/17464

 Title: Nonlinear Multicontrast Microscopy for Structural and Dynamic Investigations of Myocytes Authors: Greenhalgh, Catherine Ann Advisor: Barzda, Virginijus Department: Physics Keywords: nonlinear microscopybiological imagingharmonic generationmulti-contrast microscopy Issue Date: 16-Jul-2009 Abstract: Abstract: Nonlinear multicontrast microscopy is established in this study as an important tool for understanding biological structure and function of muscle cells. Second harmonic generation, third harmonic generation and multi-photon excitation fluorescence are acquired simultaneously in order to establish the origin of nonlinear signal generation in myocytes, and investigate myocyte structure and functionality during muscle contraction. Using structural cross-correlation image analysis, an algorithm developed specifically for this research, for the first time, third harmonic generation is shown to originate from the mitochondria in myocytes. The second harmonic, which is generated from the anisotropic bands of the sarcomeres, is further shown to be dependent on the crystalline order of the sarcomeres, thereby providing a potential diagnostic tool to evaluate disorder in muscle cells. The combination of the second and third harmonic provides complementary information that can be used to further elucidate the basic principles of muscle contraction. Time-lapse nonlinear microscopic imaging showed structural and functional dynamics in the myocytes. The second harmonic contrast revealed nonsynchronized nanocontractions of sarcomeres in relaxed, non-contracting, cardiomyocytes and Drosophila muscle samples, providing insight into the asynchronous behaviour of individual sarcomeres. Furthermore, macrocontracting samples were found to exhibit a synchronization of nanocontractions, providing new evidence for how muscles contract. Dynamic image correlation analysis, another algorithm developed specifically for this investigation, is used to reveal networks of mitochondria, which show fluctuations of multi-photon excitation fluorescence and third harmonic generation signals. The intensity fluctuations in the networks reveal both slow and fast dynamics; phase shifts of the slow dynamics between different networks are observed. Fast dynamics appear only in the inner networks, suggesting functional difference between interfibrillar and subsarcolemma mitochondria. The groundwork for studying bioenergetics of mitochondria in cardiomyocytes with nonlinear multimodal microscopy is fully developed in this work. The origin of the nonlinear signals and the development of the image analysis techniques provide a solid foundation to further study of muscle contractility and bioenergetics. URI: http://hdl.handle.net/1807/17464 Appears in Collections: DoctoralDepartment of Physics - Doctoral theses

Files in This Item:

File Description SizeFormat
Greenhalgh_Catherine_A_200903_PhD_video1.mpgSupplementary media file 13.85 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video2.mpgSupplementary media file 2775.03 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video3.mpgSupplementary media file 31.29 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video4.mpgSupplementary media file 4401.95 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video5.mpgSupplementary media file 53.44 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video6.mpgSupplementary media file 6832.21 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video7.mpgSupplementary media file 75.91 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video8.mpgSupplementary media file 85.86 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video9.mpgSupplementary media file 95.88 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video10.mpgSupplementary media file 102.94 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video11.mpgSupplementary media file 11411.42 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video12.mpgSupplementary media file 122.67 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video13.mpgSupplementary media file 13736.57 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video14.mpgSupplementary media file 14687.49 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video15.mpgSupplementary media file 151.31 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video16.mpgSupplementary media file 16853.43 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video17.mpgSupplementary media file 17522.79 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video18.mpgSupplementary media file 18356.03 kBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video19.mpgSupplementary media file 191.02 MBMPEG
View/Open
Greenhalgh_Catherine_A_200903_PhD_video20.mpgSupplementary media file 20864.01 kBMPEG
View/Open