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Confocal laser scanning microscopy—a powerful tool in bone research

Konfokale Laser-Scanning-Mikroskopie – eine leistungsfähige Methode in der Knochenforschung

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Summary

The confocal laser scanning microscope (CLSM) enables the collection of images picturing selected planes in depth of thick samples, thus giving 3D information while keeping the sample intact. In this article we give an overview of our CLSM applications in bone research: (i) the characterization of osteoblasts and osteoclasts properties in cell biology, (ii) the visualization of the three dimensional (3D) osteocyte lacunar canalicular network in undemineralized plastic-embedded bone samples, (iii) the observation of tetracycline labels in bone biopsy samples from patients in combination with information on the mineralization density from quantitative backscatter electron imaging, which enables the time course of mineral accumulation in newly formed bone to be followed, (iv) the precise measurement of the thickness of thin ground bone sections, a prerequisite for the mapping of local mechanical properties by scanning acoustic microscopy.

Zusammenfassung

Das konfokale Laser-Scanning-Mikroskop (CLSM) ermöglicht die Serienaufnahme von Bildern auch aus tiefer gelegenen Schichten einer Probe und damit 3‑D-Informationen, ohne dabei die Probe zu zerstören. Im vorliegenden Artikel geben die Autoren einen Überblick über ihre Anwendungen des CLSM in der Knochenforschung: (i) die Charakterisierung von Eigenschaften der Osteoblasten und Osteoklasten in der Zellbiologie; (ii) die dreidimensionale (3-D-)Darstellung und Vermessung des Osteozytenlakunen-Canaliculi-Netzwerks von in Kunststoff eingebetteten nichtdemineralisierten Knochenproben; (iii) die Identifikation von Tetrazyklinmarkierungen in Knochenbiopsien von Patienten, die – in Kombination mit Informationen über die lokale Kalziumkonzentration aus quantitativer Rückstreuelektronenmikroskopie – Rückschlüsse über den zeitabhängigen Anstieg der Kalziumkonzentration in neu angebautem Knochen erlaubt; (iv) die präzise Messung der Dicke von Knochendünnschliffen. Letztere Information ist die Voraussetzung für die Ermittlung bestimmter lokaler mechanischer Eigenschaften mit Ultraschallmikroskopie.

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Acknowledgements

This paper is dedicated to ao. Univ.-Prof. Dr. Adolf Ellinger, who—in his role as a consultant for microscopy—supported the strategic development plan for the foundation of the Ludwig Boltzmann Institute of Osteology (LBIO) in the late 1980s. It was his advice to acquire a CLSM as basic equipment for LBIO, at that time an innovative instrument. In the meantime, the CLSM has become a standard equipment for bone research and has been applied to a number of research questions in bone biology and material science by LBIO scientists.

The authors also thank their colleagues, in particular those who were engaged with development of the staining and imaging protocol at the outset of CLSM imaging at the LBIO.

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Authors and Affiliations

  1. Ludwig Boltzmann Institute of Osteology at the Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1st Medical Department, Hanusch Hospital, Vienna, Austria

    Stéphane Blouin, Andreas Roschger, Franz Varga, Barbara Misof, Silvia Spitzer, Paul Roschger & Klaus Klaushofer

  2. Department of Biomaterials, Max Planck Institute of Colloids and Interfaces, Postdam, Germany

    Andreas Roschger

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  1. Stéphane Blouin
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  2. Andreas Roschger
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  3. Franz Varga
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  4. Barbara Misof
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  5. Silvia Spitzer
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  6. Paul Roschger
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  7. Klaus Klaushofer
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Correspondence to Stéphane Blouin.

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Conflict of interest

S. Blouin, A. Roschger, F. Varga, B. Misof, S. Spitzer, P. Roschger, and K. Klaushofer declare that they have no competing interests.

Additional information

This article is dedicated to Professor Ellinger who recognized early the potential of CLSM and encouraged our institute to purchase such a device. In the meantime, CLSM is used in a wide range of applications at the Ludwig Boltzmann Institute of Osteology.

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Blouin, S., Roschger, A., Varga, F. et al. Confocal laser scanning microscopy—a powerful tool in bone research. Wien Med Wochenschr 168, 314–321 (2018). https://doi.org/10.1007/s10354-018-0639-x

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  • DOI: https://doi.org/10.1007/s10354-018-0639-x

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