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Probe-based confocal laser endomicroscopy (pCLE): a preclinical investigation of the male genital tract

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Abstract

The aim of this study was to assess the potential of probe-based confocal laser endomicroscopy (pCLE) as a new diagnostic imaging technique for the male genital tract. For this purpose, testes, epididymides, and vasa deferentia were obtained during transsexual surgery of healthy patients (n = 10, 26–52 years). Prior to this, testes of rats (n = 10, Sprague–Dawley) and mice (n = 8, wild-type) were examined. Ex vivo tissues were investigated by pCLE after topical fluorescence staining. Images and pCLE real-time video sequences were compared to images acquired by confocal laser scanning microscopy (CLSM); this allowed the identifying of corresponding microstructures. Interestingly, the seminiferous tubules of transsexual humans contained mainly spermatogonia due to long-term estrogen treatment, whereas the seminiferous tubules of the murine and rat spermatogenesis-related cell types were differentiated. Mosaicking improved the inspection potential by wide-angle views. Similarly, the microarchitecture of the epididymis and the vas deferens was successfully visualized in situ and on a cellular level by pCLE. In summary, pCLE allows for real-time identification of relevant microstructures responsible for spermatogenesis under ex vivo conditions. Additionally, pCLE enabled to localize vital spermatozoa in the testis thus opening up new ways to improve sperm retrieval rates during assisted reproduction. Both clinically relevant experiences hold promise to introduce this diagnostic method into a clinical study, and to investigate its potential as a clinical diagnostic procedure to expedite and improve the medical situation.

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Acknowledgments

We thank Mr. Myles Leavy for final proof reading.

Author information

Authors and Affiliations

  1. Department of Urology, Klinikum Grosshadern, University of Munich, Marchioninistr. 15, 81377, Munich, Germany

    Matthias Trottmann, Armin J. Becker, Christian G. Stief & Sabine Kölle

  2. LIFE Centre, Klinikum Grosshadern, University of Munich, Feodor-Lynen-Str. 19, 81377, Munich, Germany

    Ronald Sroka & Herbert Stepp

  3. Department of Urogenital Surgery, Clinics for Surgery Munich-Bogenhausen, Denninger Str. 15, 81679, Munich, Germany

    Bernhard Liedl

  4. Health Sciences Centre, UCD School of Medicine & Medical Science, University College Dublin, Belfield, Dublin 4, Ireland

    Sabine Kölle

Authors
  1. Matthias Trottmann
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  2. Ronald Sroka
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  3. Herbert Stepp
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  4. Bernhard Liedl
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  5. Armin J. Becker
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  6. Christian G. Stief
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  7. Sabine Kölle
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Corresponding author

Correspondence to Matthias Trottmann.

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Ethical statement

This study was approved by the Ethics Committee of the University of Munich (project number 161-12). The use of the dead animals had been approved by the Institutional Animal Care and Use Committee (IACUC).

Conflict of interest

No competing financial interests exist.

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Movie 1

Real-time view of a human testicular tubule mainly containining spermatogonia (staining: AC). (MPG 1132 kb)

Movie 2

Visualization of manipulations in human testicular tubules by pCLE. The contents of a tubulus contortus is pressed out which is recorded in real-time (staining: FITC). (MP4 434 kb)

Movie 3

In situ localization of spermatozoa in the human epididymis (see circular line, staining: FA). (MP4 2336 kb)

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Trottmann, M., Sroka, R., Stepp, H. et al. Probe-based confocal laser endomicroscopy (pCLE): a preclinical investigation of the male genital tract. Lasers Med Sci 31, 57–65 (2016). https://doi.org/10.1007/s10103-015-1828-0

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  • DOI: https://doi.org/10.1007/s10103-015-1828-0

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