Combining confocal and BSE SEM imaging for bone block surfaces
1 Anatomy Dept., University College London, UK
2 Centre for Oral Growth and Development, Barts and The London School of Medicine and Dentistry, Queen Mary, University of London, Turner St., London, UK
3 Faculty of Mechanical Engineering, Brno University of Technology, Brno, Czech Republic
Abstract
The present report presents a method for the correlation of qualitative and quantitative BSE SEM imaging with confocal scanning light microscopy (CSLM) imaging modes applied to bone samples embedded in PMMA. The SEM has a proper digital scan generator: we leave the BSE image unchanged, and match the CSLM image to it, because the CSLM scan mechanism is not digital, though the signal is digitised. Our overlapping program uses a linear transformation matrix which projects one system to the other, calculated by finding three corresponding points in BSE and CSLM pictures.BSE images are empty where cells and osteoid are present. Fluorescence mode CSLM fills in these gaps. The combination images enhance our understanding of what is going on - and re-establish the need for good cellular preservation.
Keywords
- Bone
- scanning electron microscopy
- confocal optical microscopy
- block face microscopy
- osteoid
- osteoblasts
- osteocytes
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