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Related Concept Videos

Scanning Electron Microscopy01:07

Scanning Electron Microscopy

A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a two-dimensional manner. Typically, areas between ~1 centimeter to 5 micrometers in width can be imaged. SEM can be used to image bacteria, viruses, tissues as well as larger samples like insects. Conventional SEM gives a magnification ranging from 20X to 30,000X and spatial resolution of 50 to 100 nanometers.
Fundamental Principles
Accelerated...
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
Gross Anatomy of Bone01:17

Gross Anatomy of Bone

The two main features of a long bone are the diaphysis and the epiphysis.
The diaphysis is the tubular shaft that runs between the proximal and distal ends of the bone. The walls of the diaphysis are composed of dense and hard compact bone made of numerous osteons — the functional unit of the compact bone. The hollow region in the diaphysis is called the medullary cavity, which harbors the bone marrow. In infants and children, this marrow cavity is filled with red marrow, whereas in adults, it...
Overview of Electron Microscopy01:25

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
Bone Structure01:55

Bone Structure

Within the skeletal system, the structure of a bone, or osseous tissue, can be exemplified in a long bone, like the femur, where there are two types of osseous tissue: cortical and cancellous.

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Multimodal Approach to Assess Bone Regeneration and Scaffold Performance
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Scanning electron microscopy of bone.

Alan Boyde1

  • 1Biophysics Section, Oral Growth and Development, Dental Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, UK. a.boyde@qmul.ac.uk

Methods in Molecular Biology (Clifton, N.J.)
|December 2, 2011
PubMed
Summary
This summary is machine-generated.

This chapter details Scanning Electron Microscopy (SEM) methods for bone imaging. It highlights Backscattered Electron (BSE) imaging and sample preparation techniques for detailed bone and cell visualization.

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Area of Science:

  • Bone biology and histology
  • Microscopy techniques
  • Materials science

Background:

  • Scanning Electron Microscopy (SEM) is crucial for visualizing bone microstructures.
  • Effective sample preparation is essential for high-resolution imaging of bone and cells.
  • Various SEM imaging modes offer complementary information about bone tissue.

Purpose of the Study:

  • To describe detailed methods for Scanning Electron Microscopical (SEM) imaging of bone and bone cells.
  • To provide guidance on sample preparation for optimal imaging results.
  • To highlight the utility of different SEM modes and related techniques.

Main Methods:

  • Sample preparation including fixation, drying, maceration (using alkaline bacterial pronase, hypochlorite, hydrogen peroxide, sodium/potassium hydroxide), and coating.
  • Imaging techniques such as Backscattered Electron (BSE), Secondary Electron (SE), Energy Dispersive X-ray (EDX), Cathodoluminescence (CL), and Environmental SEM (ESEM).
  • Resin embedding (including PMMA) and casting for 3D imaging, correlated imaging with confocal microscopy, microradiography, and microtomography.

Main Results:

  • Backscattered Electron (BSE) imaging is identified as the most useful SEM mode for bone research.
  • Detailed protocols are provided for preparing unembedded and resin-embedded bone samples, including maceration techniques.
  • Recommendations are given for resin embedding suitable for BSE imaging and for creating spatial casts.
  • Environmental SEM (ESEM) is effective for overcoming charging issues in cancellous bone samples.

Conclusions:

  • Optimized SEM methods, particularly BSE imaging, enable detailed 3D visualization of bone and bone cells.
  • Effective sample preparation and choice of imaging mode are critical for successful bone SEM analysis.
  • Correlating SEM with other imaging modalities enhances the comprehensive understanding of bone structure and mineralization.