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Proceedings Paper

Functional imaging of muscle cells by second harmonic generation
Author(s): Valentina Nucciotti; Leonardo Sacconi; Marco Linari; Vincenzo Lombardi; Gabriella Piazzesi; Nicoletta Piroddi; Corrado Poggesi; Chiara Tesi; Francesco Vanzi; Francesco S. Pavone
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Paper Abstract

The intrinsically ordered arrays of proteins (mainly actin and myosin) constituting the myofibrils within muscle cells are at the basis of a strong Second Harmonic Generation (SHG) from muscle fibers and isolated myofibrils. We have characterized the SHG signal with regard to its polarization and potential source within the muscle cell. The lateral resolution that can be achieved through SHG imaging of muscle strongly depends on sample depth. In fact, a comparison between intact muscle fibers and single myofibrils demonstrates that, whereas in both cases the alternation of dark I bands and bright A bands is visible, the contours of these bands are much better resolved in myofibrils than in fibers. Further, imaging of myofibrils revealed the presence of a darker zone in the centre of the A band. These effects of scattering by tissue on the image resolution were also studied with regard to the polarization of the SHG signal. The polarization-dependence of SHG intensity represents a powerful tool for the investigation of the structural dynamics occurring in the emitting proteins during the active cycle of muscle contraction. The prospective to perform functional studies requires a complete characterization of the effects of scattering and possibly multiple emitting populations on the measured SHG signal. Also, SHG is extremely sensitive to the degree of order present in the filament array, offering an interesting potential in the development of non-invasive tools for the diagnosis of degenerative diseases affecting skeletal muscles.

Paper Details

Date Published: 23 February 2006
PDF: 9 pages
Proc. SPIE 6089, Multiphoton Microscopy in the Biomedical Sciences VI, 60891I (23 February 2006); doi: 10.1117/12.646004
Show Author Affiliations
Valentina Nucciotti, Univ. of Florence (Italy)
Leonardo Sacconi, European Lab. for Non-linear Spectroscopy (Italy)
Marco Linari, Univ. of Florence (Italy)
Vincenzo Lombardi, Univ. of Florence (Italy)
Gabriella Piazzesi, Univ. of Florence (Italy)
Nicoletta Piroddi, Univ. of Florence (Italy)
Corrado Poggesi, Univ. of Florence (Italy)
Chiara Tesi, Univ. of Florence (Italy)
Francesco Vanzi, Univ. of Florence (Italy)
European Lab. for Non-linear Spectroscopy (Italy)
Francesco S. Pavone, European Lab. for Non-linear Spectroscopy (Italy)


Published in SPIE Proceedings Vol. 6089:
Multiphoton Microscopy in the Biomedical Sciences VI
Ammasi Periasamy; Peter T. C. So, Editor(s)

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