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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 | Source: | JOURNAL OF BIOMEDICAL OPTICS, 20 (8) | 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 | Keywords: | striated muscle; second harmonic generation; refractive index; supramolecular model | Document URI: | http://hdl.handle.net/1942/19091 | Link to publication/dataset: | https://www.researchgate.net/publication/281144233_On_the_interpretation_of_second_harmonic_generation_intensity_profiles_of_striated_muscle | ISSN: | 1083-3668 | e-ISSN: | 1560-2281 | DOI: | 10.1117/1.JBO.20.8.086010 | ISI #: | 000362516700021 | Rights: | © 2015 Society of Photo-Optical Instrumentation Engineers (SPIE) | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2016 |
Appears in Collections: | Research publications |
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Paesen et al JBO 2015.pdf | Published version | 883.96 kB | Adobe PDF | View/Open |
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