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A novel approach for assessing cardiac fibrosis using label-free second harmonic generation

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Abstract

To determine whether second harmonic generation (SHG) can be used as a novel and improved label-free technique for detection of collagen deposition in the heart. To verify whether SHG will allow accurate quantification of altered collagen deposition in diseased hearts following hypertrophic remodelling. Minimally invasive transverse aortic banding (MTAB) of mouse hearts was used to generate a reproducible model of cardiac hypertrophy. Physiological and functional assessment of hypertrophic development was performed using echocardiography and post-mortem analysis of remodelled hearts. Cardiac fibroblasts were isolated from sham-operated and hypertrophied hearts and proliferation rates compared. Multi-photon laser scanning microscopy was used to capture both two-photon excited autofluorescence (TPEF) and SHG images simultaneously in two channels. TPEF images were subtracted from SHG images and the resulting signal intensities from ventricular tissue sections were calculated. Traditional picrosirius red staining was used to verify the suitability of the SHG application. MTAB surgery induced significant hypertrophic remodelling and increased cardiac fibroblast proliferation. A significant increase in the density of collagen fibres between hypertrophic and control tissues (p < 0.05) was evident using SHG. Similar increases and patterns of staining were observed using parallel traditional picrosirius red staining of collagen. Label-free SHG microscopy provides a new alternative method for quantifying collagen deposition in fibrotic hearts.

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Acknowledgments

This work was supported by the Engineering and Physical Sciences Research Council (Grant No.: EP/I006826/1 and pump-priming award).

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Authors and Affiliations

  1. Institute of Cardiovascular and Medical Sciences, University of Glasgow, Wolfson Link building, Glasgow, G12 8QQ, UK

    Tamara P. Martin

  2. Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Hamnett building, 161 Cathedral Street, Glasgow, G4 ORE, UK

    Greg Norris, Gail McConnell & Susan Currie

Authors
  1. Tamara P. Martin
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  2. Greg Norris
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  3. Gail McConnell
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  4. Susan Currie
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Correspondence to Susan Currie.

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Martin, T.P., Norris, G., McConnell, G. et al. A novel approach for assessing cardiac fibrosis using label-free second harmonic generation. Int J Cardiovasc Imaging 29, 1733–1740 (2013). https://doi.org/10.1007/s10554-013-0270-2

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  • DOI: https://doi.org/10.1007/s10554-013-0270-2

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