A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin,...
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Transcript of A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin,...
![Page 1: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/1.jpg)
A Proposed Analysis of Collagen Morphology Using
Electron & Soft X-ray Microscopy
Amy H. Brazin, NASA ApprenticeRussell Garman, Ph.D. Candidate
Stefan Judex, Ph.D.~~~~~~~
SUNY Stony Brook Dept. of Biomedical Engineering
August 1, 2005
![Page 2: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/2.jpg)
Application to NASA Research
Analyzing collagen in bone under the following conditions:
Weightlessness-> astronauts experience this in space
Mechanical Stimuli-> the countering force of bone loss due to weightlessness
![Page 3: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/3.jpg)
Major structural protein that act as cables to strengthen body tissue
90% of body is Collagen Type I
Triple helix structure
Collagenous matrix correlates to bone fracture toughness
Wang, X., Bank, R. et al. “The Role of Collagen in Determining Bone Mechanical Properties.” Journal of Orthopedic Research. 19(2001) 1021-1026
Role of Collagen in the Body
![Page 4: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/4.jpg)
Understanding Collagen Morphology
The following may elude to the elasticity and fracture toughness of the bone:
Diameter and density of fibersOrientation
Fibril diameters and distributions are associated with:
Tissue growth and maturationTissue response to physical stress
Frank, C., Bray, D. “Electron Microscopic Quantification of Collagen Fibril Diameters in the Rabbit Medial Collateral Ligament: A Baseline for Comparison.” Connect Tissue Res.
19:11-25. 1989.
![Page 5: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/5.jpg)
Current Task at Hand
To develop a protocol to:
Prepare calcified and decalcified bones
Cut ultrathin sections using a microtome
Establish standard resolution features for microscopy work
![Page 6: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/6.jpg)
Sample Preparation for AnalysisFixation
Decalcified BonesUndecalcified Bones
Post Fixation
Dehydration
Infiltration
Embedding
Sectioning, 100nm
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![Page 7: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/7.jpg)
Analyzing Collagen
Transmission Electron MicroscopyHigh resolutions allow the quantification of:
Collagen fibrils, subfibrils and microfibrilsDiameter distribution
Soft X-ray MicroscopyDifferentiates the mineral quantities in bone as well as the crystalline properties of the bone matrices
Frank, C., Bray, D. “Electron Microscopic Quantification of Collagen Fibril Diameters in the Rabbit Medial Collateral Ligament: A Baseline for Comparison.” Connect Tissue Res.
19:11-25. 1989.
![Page 8: A Proposed Analysis of Collagen Morphology Using Electron & Soft X-ray Microscopy Amy H. Brazin, NASA Apprentice Russell Garman, Ph.D. Candidate Stefan.](https://reader036.fdocuments.in/reader036/viewer/2022083008/56649f005503460f94c15f8b/html5/thumbnails/8.jpg)
Acknowledgements
Special thanks to the following people, whom without, this research would not be possible:
Hoyan LamAdiba AliNASA SHARP