Binary imaging analysis for comprehensive quantitative histomorphometry of peripheral nerve.
Quantitative histomorphometry is the current gold standard for objective measurement of nerve architecture and its components. Many methods still in use rely heavily upon manual techniques that are prohibitively time consuming, predisposing to operator fatigue, sampling error, and overall limited reproducibility. More recently, investigators have attempted to combine the speed of automated morphometry with the accuracy of manual and semi-automated methods. Systematic refinements in binary imaging analysis techniques combined with an algorithmic approach allow for more exhaustive characterization of nerve parameters in the surgically relevant injury paradigms of regeneration following crush, transection, and nerve gap injuries. The binary imaging method introduced here uses multiple bitplanes to achieve reproducible, high throughput quantitative assessment of peripheral nerve. Number of myelinated axons, myelinated fiber diameter, myelin thickness, fiber distributions, myelinated fiber density, and neural debris can be quantitatively evaluated with stratification of raw data by nerve component. Results of this semi-automated method are validated by comparing values against those obtained with manual techniques. The use of this approach results in more rapid, accurate, and complete assessment of myelinated axons than manual techniques.
Division of Plastic and Reconstructive Surgery, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8238, Saint Louis, MO 63110, United States. hunterd@wudosis.wustl.edu
Daniel A
DA
Hunter
Arash
A
Moradzadeh
Elizabeth L
EL
Whitlock
Michael J
MJ
Brenner
Terence M
TM
Myckatyn
Cindy H
CH
Wei
Thomas H H
TH
Tung
Susan E
SE
Mackinnon
D000465:Algorithms; D000818:Animals; D001369:Axons / Q000502:physiology / Q000648:ultrastructure*; D019044:Image Cytometry / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D009186:Myelin Sheath / Q000502:physiology / Q000648:ultrastructure; D009413:Nerve Fibers, Myelinated / Q000502:physiology / Q000648:ultrastructure*; D010363:Pattern Recognition, Automated / Q000379:methods*; D010525:Peripheral Nerves / Q000166:cytology* / Q000502:physiology; D010784:Photomicrography; D051381:Rats; D011917:Rats, Inbred Lew; D012583:Schwann Cells / Q000166:cytology / Q000502:physiology; D012984:Software; D013194:Staining and Labeling / Q000379:methods