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Next-generation endomyocardial biopsy: the potential of confocal and super-resolution microscopy

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Abstract

Confocal laser scanning microscopy and super-resolution microscopy provide high-contrast and high-resolution fluorescent imaging, which has great potential to increase the diagnostic yield of endomyocardial biopsy (EMB). EMB is currently the gold standard for identification of cardiac allograft rejection, myocarditis, and infiltrative and storage diseases. However, standard analysis is dominated by low-contrast bright-field light and electron microscopy (EM); this lack of contrast makes quantification of pathological features difficult. For example, assessment of cardiac allograft rejection relies on subjective grading of H&E histology, which may lead to diagnostic variability between pathologists. This issue could be solved by utilising the high contrast provided by fluorescence methods such as confocal to quantitatively assess the degree of lymphocytic infiltrate. For infiltrative diseases such as amyloidosis, the nanometre resolution provided by EM can be diagnostic in identifying disease-causing fibrils. The recent advent of super-resolution imaging, particularly direct stochastic optical reconstruction microscopy (dSTORM), provides high-contrast imaging at resolution approaching that of EM. Moreover, dSTORM utilises conventional fluorescence dyes allowing for the same structures to be routinely imaged at the cellular scale and then at the nanoscale. The key benefit of these technologies is that the high contrast facilitates quantitative digital analysis and thereby provides a means to robustly assess critical pathological features. Ultimately, this technology has the ability to provide greater accuracy and precision to EMB assessment, which could result in better outcomes for patients.

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  1. ImageJ can be download from http://imagej.nih.gov/ij/ or http://fiji.sc/Fiji.

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Acknowledgments

We thank Jacqui Ross and Nicky Kingston for assistance with microscopy, Auckland City Hospital staff for assistance in obtaining tissue and transplant recipients, and donor families for donating tissue. Research funding was provided by the Auckland Medical Research Foundation and the Health Research Council of New Zealand.

Ethical standard

Human cardiac tissue was obtained with the written informed consent of heart transplant recipients and from families of organ donors as approved by the New Zealand Health and Disability Ethics Committee (NTY/05/08/050).

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Authors and Affiliations

  1. Department of Physiology, University of Auckland, Auckland, New Zealand

    David J. Crossman, Yu Feng Hou & Christian Soeller

  2. Green Lane Cardiovascular Service, Auckland City Hospital, Auckland, New Zealand

    Peter N. Ruygrok

  3. Biomedical Physics, University of Exeter, Exeter, UK

    Christian Soeller

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  1. David J. Crossman
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  2. Peter N. Ruygrok
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  3. Yu Feng Hou
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  4. Christian Soeller
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Correspondence to David J. Crossman.

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Crossman, D.J., Ruygrok, P.N., Hou, Y.F. et al. Next-generation endomyocardial biopsy: the potential of confocal and super-resolution microscopy. Heart Fail Rev 20, 203–214 (2015). https://doi.org/10.1007/s10741-014-9455-6

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