Legs, including femora and tibiae, were fixed in 10% buffered formalin, decalcified in 14% EDTA, and embedded in Tissue-Tek. Frozen sections (8μm thick) were cut using a Leica CM1850 cryostat (Leica, German). 1) Sections were mounted with a mounting medium containing DAPI (Vector Lab., Burlingame, CA, USA) for GFP+ tumor burden examination. 2) Adjacent sections were stained with H&E for general histology. Histomorphometric analyses were performed on sections cut at 3 representative levels in each sample using CMSR SOPs (43 (link),44 (link)). In brief, sections were numbered blindly and were converted to digital images using an Olympus VS120 whole slide imaging system (Olympus, Center Valley, PA). GFP+ areas were analyzed using Image Pro-Plus version 6.0 software (Media Cybernetics, Rockville, MD, USA). The percentage of trabecular bone area over total tissue area was analyzed using OsteoMeasure Image Analysis System (OsteoMetrics, Decatur, GA, USA).
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Histomorphometric Analysis of Tumor Burden
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Corresponding Organization : University of Rochester Medical Center
Other organizations : University of Rochester, University of Minnesota Medical Center, BioVinc (United States)
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Variable analysis
independent variables
- None explicitly mentioned
- GFP+ tumor burden
- Percentage of trabecular bone area over total tissue area
- Legs, including femora and tibiae, were fixed in 10% buffered formalin
- Samples were decalcified in 14% EDTA
- Samples were embedded in Tissue-Tek
- Sections were cut at 8μm thickness using a Leica CM1850 cryostat
- Sections were mounted with a mounting medium containing DAPI
- Adjacent sections were stained with H&E for general histology
- Histomorphometric analyses were performed on sections cut at 3 representative levels in each sample
- Sections were numbered blindly and converted to digital images using an Olympus VS120 whole slide imaging system
- GFP+ areas were analyzed using Image Pro-Plus version 6.0 software
- The percentage of trabecular bone area over total tissue area was analyzed using OsteoMeasure Image Analysis System
- Positive control: None mentioned
- Negative control: None mentioned
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