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Related Concept Videos

Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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Correction: ISSLS Prize in Basic Science 2026: Early markers of mechanical modulation in whole bovine intervertebral discs loaded in a multiaxial bioreactor.

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Staining and High-Resolution Imaging of Three-Dimensional Organoid and Spheroid Models
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Confocal imaging protocols for live/dead staining in three-dimensional carriers.

Benjamin Gantenbein-Ritter1, Christoph M Sprecher, Samantha Chan

  • 1ARTORG, Center for Biomedical Engineering Research, Institute for Surgical Technology and Biomechanics, University of Bern, Bern, Switzerland. benjamin.gantenbein@artorg.unibe.ch

Methods in Molecular Biology (Clifton, N.J.)
|April 7, 2011
PubMed
Summary

This study introduces new staining protocols for live cell imaging in thick 3D tissue engineering constructs. These methods enable confocal laser scanning microscopy for evaluating cell survival in complex scaffolds like fibrin gels and alginate beads.

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Area of Science:

  • Tissue Engineering
  • Biomaterials Science
  • Cell Biology

Background:

  • Confocal laser scanning microscopy (CLSM) is vital for live cell imaging in 3D constructs.
  • Current CLSM applications are limited to thin samples (<20 μm), hindering analysis of thicker tissues or scaffolds.
  • There's a need for improved protocols to utilize CLSM's full potential in evaluating cell viability within larger 3D environments.

Purpose of the Study:

  • To develop and present novel staining protocols for effective live cell imaging in thick 3D tissue engineering samples.
  • To enable the application of confocal laser scanning microscopy for assessing cell survival in tissue constructs and biomaterial scaffolds.
  • To overcome the limitations of current methods that restrict CLSM to analyzing only thin sample slices.

Main Methods:

  • Development of specialized staining techniques for 3D tissue samples, including intervertebral disc tissue.
  • Adaptation of staining protocols for various biomaterial scaffolds, such as fibrin gels and alginate beads.
  • Application of confocal laser scanning microscopy to visualize and evaluate cell viability within these stained 3D constructs.

Main Results:

  • Successful implementation of new protocols allowing deep tissue penetration for live cell imaging.
  • Demonstration of CLSM's capability to analyze cell survival in 3D scaffolds with thicknesses up to several millimeters.
  • Validation of the protocols across different 3D tissue and scaffold types, showing consistent efficacy.

Conclusions:

  • The presented staining protocols significantly enhance the utility of confocal laser scanning microscopy for 3D tissue engineering.
  • These advancements facilitate more accurate and comprehensive evaluation of cell survival in complex, thick biological constructs.
  • The developed methods open new avenues for research in tissue regeneration and biomaterial development by enabling detailed 3D cellular analysis.