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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/19091

Title: On the interpretation of second harmonic generation intensity profiles of striated muscle
Authors: Paesen, Rik
Smolders, Sophie
Wens, Inez
Notelaers, Kristof
de Hoyos Vega, Manolo José
Bito, Virginie
Op't Eijnde, Bert
Hansen, Dominique
Ameloot, Marcel
Issue Date: 2015
Abstract: Recently, a supramolecular model was developed for predicting striated skeletal muscle intensity profiles obtained by label-free second harmonic generation (SHG) microscopy. This model allows for a quantitative determination of the length of the thick filament antiparallel range or M-band (M), and results in M ¼ 0.12 μm for single-band intensity profiles when fixing the A-band length (A) to A ¼ 1.6 μm, a value originating from electron microscopy (EM) observations. Using simulations and experimental data sets, we showed that the objective numerical aperture (NA) and the refractive index (RI) mismatch (Δn ¼ n2ω − nω) between the illumination wave (ω) and the second harmonic wave (2ω) severely affect the simulated sarcomere intensity profiles. Therefore, our recovered filament lengths did not match with those observed by EM. For an RI mismatch of Δn ¼ 0.02 and a moderate illumination NA of 0.8, analysis of single-band SHG intensity profiles with freely adjustable A- and M-band sizes yielded A ¼ 1.40 0.04 μm and M ¼ 0.07 0.05 μm for skeletal muscle. These lower than expected values were rationalized in terms of the myosin density distribution along the myosin thick filament axis. Our data provided new and practical insights for the application of the supramolecular model to study SHG intensity profiles in striated muscle.
Notes: Ameloot, M (reprint author), Hasselt Univ, Biomed Res Inst, Agoralaan Bldg C, B-3590 Diepenbeek, Belgium. marcel.ameloot@uhasselt.be
URI: http://hdl.handle.net/1942/19091
Link to publication: https://www.researchgate.net/publication/281144233_On_the_interpretation_of_second_harmonic_generation_intensity_profiles_of_striated_muscle
DOI: 10.1117/1.JBO.20.8.086010
ISI #: 000362516700021
ISSN: 1083-3668
Category: A1
Type: Journal Contribution
Validation: ecoom, 2016
Appears in Collections: Research publications

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