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Related Experiment Videos

High temporal and spatial resolution studies of bone cells using real-time confocal reflection microscopy

A Boyde1, P Vesely, C Gray

  • 1Department of Anatomy and Developmental Biology, University College London, England.

Scanning
|September 1, 1994
PubMed
Summary

High-speed confocal microscopy reveals rapid organelle movement within bone cells like osteoclasts and osteoblasts. Different microscope techniques were evaluated for studying live bone cells over extended periods.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Understanding the dynamic behavior of bone cells (osteoclasts and osteoblasts) is crucial for bone physiology and pathology.
  • High-speed microscopy techniques are needed to capture rapid intracellular events in living bone cells.
  • Confocal microscopy offers optical sectioning capabilities essential for detailed cellular analysis.

Purpose of the Study:

  • To evaluate and compare different high-speed scanning confocal microscopy techniques for studying live bone-derived cells.
  • To document the real-time intracellular dynamics, specifically organelle movement, within osteoclasts and osteoblasts.
  • To assess the suitability of these advanced microscopy methods for long-term live-cell imaging.

Main Methods:

  • Utilized three types of high-speed scanning confocal microscopes: Noran Tandem Scanning Microscope (TSM), Noran Odyssey video rate laser confocal scanning microscope (VRCSLM), and Lasertec 1LM11 line scan instrument.

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  • Studied chick and rat bone-derived osteoclasts and osteoblasts at 37°C in sealed chambers.
  • Employed techniques including reflection confocal imaging, phase contrast, and interference reflection modes, with rapid focus adjustment and time-lapse movie sequences.

    • Main Results:

    • All microscopy methods demonstrated rapid net movement of organelles within bone cells.
    • Video rate laser confocal scanning microscopes (VRCSLMs) effectively documented rapid, real-time oscillatory motion.
    • Interference reflection mode provided strong contrasts in confocal instruments, and TSM allowed simultaneous use of phase contrast.

    Conclusions:

    • High-speed scanning confocal microscopy, particularly VRCSLMs, is well-suited for observing rapid intracellular dynamics in live bone cells.
    • The evaluated methods allow for extended observation of bone cells (up to 5 days) with minimal apparent irradiation damage.
    • These advanced imaging techniques provide valuable insights into the real-time behavior of osteoclasts and osteoblasts.