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Non-invasive Optical Imaging of the Lymphatic Vasculature of a Mouse
Published on: March 8, 2013
Quantification of microvessels in canine lymph nodes
Zbynĕk Tonar1, Gunter F Egger, Kirsti Witter
1Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague, Karlovarska 48, 301 66 Pilsen, Czech Republic. tonar@ntc.zcu.cz
Microscopy Research and Technique
|July 11, 2008
Summary
A new unbiased method for quantifying tumor microvessels improves reproducibility and reduces observer variance. This systematic sampling approach offers a more reliable alternative to traditional hot spot counting in canine lymph nodes.
Area of Science:
- Veterinary Pathology
- Oncology
- Stereology
Background:
- Microvessel quantification in tumors often relies on subjective hot spot analysis.
- This traditional method suffers from poor reproducibility and significant interobserver variability.
- Variability stems from inconsistent sampling of image fields for analysis.
Purpose of the Study:
- To evaluate an unbiased stereological method for quantifying microvessels in canine lymph nodes.
- To compare this unbiased method with traditional hot spot counting.
- To assess the inter- and intraobserver variability of the unbiased method.
Main Methods:
- Systematic uniform random sampling of 50 image fields per lymph node.
- Immunohistochemistry (von Willebrand factor) and lectin histochemistry (wheat germ agglutinin) for endothelium detection.
- Application of an unbiased counting frame and stereological counting rules.
Main Results:
- The unbiased method yielded microvessel density estimates ranging from 100.5 to 442.6/mm(2).
- Hot spot counting produced higher, more variable estimates (264 to 771/mm(2)).
- The unbiased method demonstrated good reproducibility with low inter- and intraobserver variance.
Conclusions:
- Systematic uniform random sampling provides a reproducible and less variable method for microvessel quantification.
- This unbiased approach is suitable for analyzing archival tissue and excluding artifacts.
- While effective, the unbiased method is more time-consuming than hot spot analysis.
