Measurement of muscle disease by quantitative second-harmonic generation imaging

University of Connecticut Health Center, Department of Genetics and Developmental Biology, Farmington, Connecticut 06030, USA.
Journal of Biomedical Optics (Impact Factor: 2.86). 01/2008; 13(4):044018. DOI: 10.1117/1.2967536
Source: PubMed


Determining the health of muscle cells by in vivo imaging could impact the diagnosis and monitoring of a large number of congenital and acquired muscular or cardiac disorders. However, currently used technologies are hampered by insufficient resolution, lack of specificity, or invasiveness. We have combined intrinsic optical second-harmonic generation from sarcomeric myosin with a novel mathematical treatment of striation pattern analysis, to obtain measures of muscle contractile integrity that correlate strongly with the neuromuscular health of mice suffering from genetic, acquired, and age-related decline in skeletal muscle function. Analysis of biopsies from a pilot group of human volunteers suggests a similar power in quantifying sarcopenic changes in muscle integrity. These results provide the first strong evidence that quantitative image analysis of sarcomere pattern can be correlated with physiological function, and they invite the application of SHG imaging in clinical practice, either in biopsy samples or via microendoscopy.


Available from: Sergey Plotnikov, Nov 12, 2014
  • Source
    • "reveals the overall cell delineation. Recently, muscle disease was quantified with SHG microscopy images [54]. Parameters, such as distribution of lengths of sarcomeres and muscle fiber crosssectional areas, were measured. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Current research on multiphoton autofluorescence microscopy is primarily focused on imaging the signal from reduced nicotinamide adenine dinucleatide (NADH) in tissue. NADH levels in cells are useful reporters of metabolic information, as well as early indicators in precancer and cancer diagnosis. While NADH is typically imaged in the 400-500 nm spectral window, the amino acid tryptophan is the major source of tissue fluorescence in the Ultraviolet range. Here, we briefly review current progress in multiphoton autofluorescence imaging of live tissues and cells, and report our recent findings of in vivo mouse skin imaging based on multiphoton excited tryptophan autofluorescence. This new method enables noninvasive imaging of skin tissue at video-rate and allows for the visualization and identification of cellular components in the epidermis, dermis, and muscle layers. It is also possible to image through small blood vessels in the mouse skin and observe circulating leukocytes in situ .
    IEEE Journal of Selected Topics in Quantum Electronics 07/2010; 16(3-16):516 - 523. DOI:10.1109/JSTQE.2009.2031619 · 2.83 Impact Factor
  • Source
    • "Polarization Second Harmonic Microscopy (PSHM) is an emerging tool that provides a new dimension to Second Harmonic imaging. Measurement and subsequent analysis of polarization dependency of Second Harmonic (SH) allows retrieving quantitative and/or subdiffraction information about molecular architecture of the imaged specimen[5] [6] [7] [8] [9]. Particularly, statistical distribution of myosin helical pitch angle [6] [7] can be estimated in different tissues, or in more practical terms, a biomarker which quantifies a mixture of subdiffraction information related to active SHG structures. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Fetal growth restriction (FGR) has recently shown a strong association with cardiac programming which predisposes to cardiovascular mortality in adulthood. Polarization Second Harmonic Microscopy can quantify molecular architecture changes with high sensitivity in cardiac myofibrils. In this work, we use myosin helical pitch angle as an example to quantify such alterations related to this high risk population. Importantly, this shows a potential use of the technique as an early diagnostic tool and an alternative method to understand pathophysiological processes.
    Proceedings of SPIE - The International Society for Optical Engineering 06/2009; DOI:10.1117/12.831481 · 0.20 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Sarcomeres are the basic contractile units of striated muscle. Our knowledge about sarcomere dynamics has primarily come from in vitro studies of muscle fibres and analysis of optical diffraction patterns obtained from living muscles. 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 microendoscopy to observe second-harmonic frequencies of light generated in the muscle fibres of live mice and humans. Using microendoscopes as small as 350 microm 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.
    Nature 08/2008; 454(7205):784-8. DOI:10.1038/nature07104 · 41.46 Impact Factor
Show more