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Minimally invasive high-speed imaging of sarcomere contractile dynamics in mice and humans

Abstract

Sarcomeres are the basic contractile units of striated muscle. Our knowledge about sarcomere dynamics has primarily come from in vitro studies of muscle fibres1 and analysis of optical diffraction patterns obtained from living muscles2,3. Both approaches involve highly invasive procedures and neither allows examination of individual sarcomeres in live subjects. Here we report direct visualization of individual sarcomeres and their dynamical length variations using minimally invasive optical microendoscopy4 to observe second-harmonic frequencies of light generated in the muscle fibres5,6 of live mice and humans. Using microendoscopes as small as 350 μm in diameter, we imaged individual sarcomeres in both passive and activated muscle. Our measurements permit in vivo characterization of sarcomere length changes that occur with alterations in body posture and visualization of local variations in sarcomere length not apparent in aggregate length determinations. High-speed data acquisition enabled observation of sarcomere contractile dynamics with millisecond-scale resolution. These experiments point the way to in vivo imaging studies demonstrating how sarcomere performance varies with physical conditioning and physiological state, as well as imaging diagnostics revealing how neuromuscular diseases affect contractile dynamics.

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Figure 1: Minimally invasive microendoscopy system.
Figure 2: Static imaging of individual sarcomeres.
Figure 3: Dynamic imaging of mouse sarcomeres.
Figure 4: Dynamic imaging of human sarcomeres.

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Acknowledgements

We thank M. Cromie and B. Flusberg for technical assistance, M. Cromie and A. Lewis for assistance with the human subject protocol, D. Profitt for expert machining, and Y. Goldman, R. Lieber, F. Zajac and T. Sanger for discussions. This work was supported by the Coulter Foundation (S.L.D. and M.J.S.), a Stanford Bio-X Interdisciplinary Initiatives award (S.L.D. and M.J.S.), NINDS R01NS050533 (M.J.S.), the Stanford-NIH Medical Scientist Training Program (M.E.L.), the Stanford-NIH Biophysics Training Grant (R.P.J.B.), and an equipment donation from Prairie Technologies, Inc. (M.J.S.).

Author Contributions All authors designed the experiments and interpreted data. M.E.L. and R.P.J.B collected data. M.E.L. performed the analysis. All authors discussed the results and contributed to the text.

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Correspondence to Mark J. Schnitzer.

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This research involved the use of equipment that was a gift to Stanford University from Prairie Technologies. Stanford University has filed patent applications based on part of the work documented in this manuscript.

Supplementary information

Supplementary Information 1

This file contains Supplementary Figure 1 and Legend, Supplementary Video 1 Legend, and Supplementary Methods with additional references. (PDF 766 kb)

Supplementary Video

This file contains Supplementary Video 1 which presents a three-dimensional stack of images acquired by second-harmonic microendoscopy from the lateral gastrocnemius muscle of a living mouse. (MOV 5779 kb)

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Llewellyn, M., Barretto, R., Delp, S. et al. Minimally invasive high-speed imaging of sarcomere contractile dynamics in mice and humans. Nature 454, 784–788 (2008). https://doi.org/10.1038/nature07104

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