Three-sectioning method: A procedure for studying hard tissues and large pieces under light and electron microscopy.

Area of Science:

• Histology
• Materials Science
• Biomedical Engineering

Background:

• Histological studies of hard tissues like tendons and calcified lesions are increasingly important due to advancements in implantable prostheses.
• Serial sectioning is crucial for identifying areas of interest for microscopy in complex tissue samples.

Purpose of the Study:

• To introduce an efficient "three-sectioning method" for localizing and studying areas of interest in hard histological samples.
• To facilitate correlative microscopy across different magnifications and techniques.

Main Methods:

• The proposed method involves three cycles of embedding and sectioning hard tissue samples.
• Initial thick sections (approx. 300 μm) are prepared, followed by re-embedding and sectioning into semithin sections (1-3 μm).
• Further re-embedding and sectioning into ultrathin sections (60-90 nm) are performed for transmission electron microscopy, if needed.

Main Results:

• The method enables the selection and detailed study of specific areas of interest within hard tissues.
• It integrates light microscopy and electron microscopy techniques seamlessly.
• The technique supports correlative microscopy from low to high magnifications.

Conclusions:

• The "three-sectioning method" provides an efficient and systematic approach for histological analysis of hard tissues.
• This technique enhances the application of various microscopy techniques, including scanning and transmission electron microscopy.
• It is a valuable tool for research involving implantable prostheses and other hard tissue studies.