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A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT
Published on: June 12, 2020
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Chemical etching in processing cortical bone specimens for scanning electron microscopy
Terenzio Congiu1, Ugo E Pazzaglia, Petra Basso
1Dipartimento di Morfologia Umana, Università dell'Insubria, 21100, Varese, Italy.
Microscopy Research and Technique
|May 28, 2014
Summary
Chemical etching of bone sections is crucial for imaging. Weak acid or polyphosphate treatments enhance osteon lamellar structure visibility without damaging collagen, unlike strong etchants.
Area of Science:
- Biomaterials Science
- Skeletal Biology
- Materials Science
Background:
- Undecalcified cortical bone sectioning is vital for studying osteon lamellar structure.
- Grinding and polishing create debris that ultrasonication alone cannot remove.
- Chemical etching is used to clean and enhance bone surfaces for microscopy.
Purpose of the Study:
- To investigate the effects of various chemical etching solutions on undecalcified human cortical bone surfaces.
- To compare the impact of different etchants, pH levels, concentrations, and contact times on SEM imaging.
- To determine optimal etching conditions for visualizing the lamellar bone pattern.
Main Methods:
- Cortical bone specimens from human metatarsals were prepared.
- Cut surfaces were treated with various acid and basic etching solutions under controlled conditions.
- Specimen surfaces were analyzed using scanning electron microscopy (SEM).
Main Results:
- Strong acid or basic etching caused significant decalcification and collagen matrix damage, hindering analysis.
- Weak acid etching (citric, acetic acid) enhanced lamellar pattern visibility but caused alterations.
- Polyphosphate treatment effectively decalcified a thin layer, improving fibril visibility without significant distortion.
Conclusions:
- Etching parameters critically influence SEM imaging of bone microstructures.
- Weak acid and polyphosphate etching offer superior results for visualizing lamellar bone compared to strong etchants.
- Careful selection of etchants is essential for accurate morphological and morphometric analysis of bone samples.
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