Osteomeasure image analysis system

Manufactured by OsteoMetrics

Sourced in United States

The OsteoMeasure Image Analysis System is a software-based tool designed to analyze digital images of bone and cartilage. The system provides quantitative measurements and data related to the morphological characteristics of these tissues.

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Lab products found in correlation

Mca771g, by Bio-Rad (1 mentions) Anti cd3, by Leica (1 mentions) Histoquest software, by TissueGnostics (1 mentions) Vs 120 whole slide imaging system, by Olympus (1 mentions) Cm1850 cryostat, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Image pro plus version 6, by Media Cybernetics (1 mentions) Xylenol orange, by Merck Group (1 mentions) Calcein, by Merck Group (1 mentions) Rm2155 hard tissue microtome, by Leica (1 mentions) Dm5500 system, by Leica (1 mentions) Rm2255, by Leica (1 mentions) Vs120, by Olympus (1 mentions) Vivact 40, by Scanco (1 mentions) Tetrachrome counter stain, by Polysciences (1 mentions)

9 protocols using osteomeasure image analysis system

Histopathological Analysis of Murine Joints

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Hind paws were prepared and analyzed by using previously described histopathologic techniques [32 (link)-35 (link)]. Staining with H&E allowed a general assessment, and toluidine blue (TB) destaining was performed to determine cartilage matrix loss. Tartrate-resistant acid phosphatase (TRAP) staining was performed to identify osteoclasts. Histomorphometric parameters (area of cartilage destruction, inflammation and erosion, as well as osteoclast numbers) were quantified by using the OsteoMeasure™ image analysis system (OsteoMetrics, Decatur, GA, USA).
Additional immunohistochemistry was done for T cells (anti-CD3; Novocastra Laboratories, Newcastle upon Tyne, UK), B cells (anti-CD45 receptor; BD Biosciences PharMingen, San Diego, CA, USA), macrophages (clone F4/80; AbD Serotec, Puchheim, Germany) and granulocytes (MCA771G; AbD Serotec) as reported previously [33 (link)-35 (link)], followed by quantitative analysis of the inflammatory infiltrate by tissue cytometry using HistoQuest™ software (TissueGnostics, Vienna, Austria) [36 (link),37 (link)].

Hadaschik E.N., Wei X., Leiss H., Heckmann B., Niederreiter B., Steiner G., Ulrich W., Enk A.H., Smolen J.S, & Stummvoll G.H. (2015). Regulatory T cell-deficient scurfy mice develop systemic autoimmune features resembling lupus-like disease. Arthritis Research & Therapy, 17(1), 35.

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Histomorphometric Analysis of Tumor Burden

Cited 2 times

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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).

Tao J., Srinivasan V., Yi X., Zhao Y., Zhang H., Lin X., Zhou X., Boyce B.F., Villalta P.W., Ebetino F.H., Ho K.K., Boeckman RK J.r, & Xing L. (2022). Bone-targeted Bortezomib Inhibits Bortezomib-Resistant Multiple Myeloma in Mice by Providing Higher Levels of Bortezomib in Bone. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 37(4), 629-642.

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Comprehensive Bone Histomorphometry Analysis

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Bone histomorphometry analysis was performed on the L3-L5 vertebrae, as previously described 25 (link), 26 . Femur sections were processed in Periodate- Lysine-Paraformaldehyde Fixative (TIANDZ) for 24h. A Shandon Finesse ME microtome was used to section unstained femur sections that were 4 µm thick before they were examined under fluorescence microscopy prior to calcein double labeling analysis. The average width along with the labeling period (days between injections of 10 μg/g calcein intraperitoneally at 7 days and 2 days before euthanasia) was then used to calculate the mineralizing surface (MS/BS, %), bone formation rate (BFR, μm³μm^-2 per day) and mineral apposition rate (MAR, μm per day).
L3-L5 vertebrae were fixed in 4% paraformaldehyde for 48 h and subjected to 48 hours of exposure to 70% ethanol before being decalcified in 10% EDTA for 14-21 days. The processed samples were finally paraffin-embedded, sectioned at a thickness of 4 µm, and stained by hematoxylin-eosin staining (H&E) and tartaric acid-resistant acid phosphatase staining (TRAP). The bone histomorphometry parameters, including osteoblast surfaces (Ob.S/BS, %), number of osteoclasts (N.Oc/B.Pm, /mm) and osteoclast surfaces (Oc.S/BS, %), were analyzed by an investigator blinded to sample collection and group assignment using an OsteoMeasure Image Analysis System (Osteometrics, Decatur, GA).

Shen G., Ren H., Shang Q., Zhang Z., Zhao W., Yu X., Tang J., Yang Z., Liang D, & Jiang X. (2020). miR-128 plays a critical role in murine osteoclastogenesis and estrogen deficiency-induced bone loss. Theranostics, 10(10), 4334-4348.

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Quantifying Bone Formation Dynamics

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Calcein (10 mg kg⁻¹) and xylenol orange (90 mg kg⁻¹) (both from Sigma‐Aldrich, St. Louis, MO, USA) were injected subcutaneously on days 13 and 3 before sacrifice, respectively. After termination, femurs were fixed in 4% paraformaldehyde solution, followed by dehydration using gradient ethanol, vitrification by xylene and embedded in methyl methacrylate. The bones were cut coronally into 10 µm thick sections with the RM2155 hard tissue microtome (Leica, Wetzlar, Germany). The double labeling images of bone slices were captured using a fluorescent microscope (Leica DM5500 system, Germany). Cortical bone formation was analyzed in four randomly selected visual fields in the midshaft of femur. Mineral apposition rate (MAR) was analyzed by OsteoMeasure Image Analysis System (Osteometrics, Decatur, GA, USA).

Wang H., Lin S., Feng L., Huang B., Lu X., Yang Z., Jiang Z., Li Y., Zhang X., Wang M., Wang B., Kong L., Pan Q., Bai S., Li Y., Yang Y., Lee W.Y., Currie P.D., Lin C., Jiang Y., Chen J., Tortorella M.D., Li H, & Li G. (2023). Low‐Dose Staphylococcal Enterotoxin C2 Mutant Maintains Bone Homeostasis via Regulating Crosstalk between Bone Formation and Host T‐Cell Effector Immunity. Advanced Science, 10(28), 2300989.

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Femur Decalcification and Osteoclast Analysis

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The right femurs were decalcified at room temperature in 10% buffered ethylenediaminetetraacetic acid for 2 weeks. Then, the bones were dehydrated using gradient ethanol, vitrified by xylene, and embedded in paraffin. 5 µm thick sections were prepared with a rotary microtome (RM2255; Leica Microsystems, Germany), then followed by TRAP staining and counterstained with Fast Green. For the analysis of osteoclasts, five fields of view in the distal femur were randomly selected for each bone slice. The Oc. N/BS, mm⁻¹ was analyzed using an OsteoMeasure Image Analysis System (Osteometrics, Decatur, GA, USA).

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Evaluation of Bone Marrow Adiposity

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For evaluation of bone marrow adiposity in proximal tibia, longitudinal sections (5 µm thick) were cut on a Jung Reichart microtome as described [60 (link)]. Measurements were performed in unstained sections under ultraviolet illumination in a sampling site located in the metaphysis, immediately below the growth plate. All measurements were performed with the OsteoMeasure Image Analysis system (OsteoMetrics, Inc., Decatur, GA, USA). Bone marrow adiposity (adipocyte area/tissue area, %), adipocyte number (number/tissue area, #/mm²), and adipocyte size (µm²) were determined as described [61 (link)].

Turner R.T., Wong C.P., Fosse K.M., Branscum A.J, & Iwaniec U.T. (2021). Caloric Restriction and Hypothalamic Leptin Gene Therapy Have Differential Effects on Energy Partitioning in Adult Female Rats. International Journal of Molecular Sciences, 22(13), 6789.

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Comprehensive Analysis of Murine Joint Structure

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For μCT, knee joints were dissected free of soft tissue, fixed overnight in 10% buffered formalin, and scanned at high resolution (10.5 μm) on a VivaCT40 μCT scanner (Scanco Medical) using 300ms integration time, 55 kVp energy, and 145 μA intensity. 3D images were generated using a constant threshold of 275 for all samples. For histology and histomorphometric analyses, knee and ankle joints were fixed in 10% buffered formalin, decalcified in 10% EDTA, embedded in paraffin, and sectioned at 4 μm thickness for 3 levels (50 μm apart). Sections were stained with H&E for routine histology and for TRAP activity to identify osteoclasts. The stained sections were digitized using a whole slide imaging system (Olympus VS120). Osteoclast numbers/mm bone surface, bone volume/tissue volume, inflamed area/tissue area and eroded surface/bone surface were analyzed on one section from each level using an osteomeasure image analysis system (Osteometrics)^{8 (link)}.

Sun W., Meednu N., Rosenberg A., Rangel-Moreno J., Wang V., Glanzman J., Owen T., Zhou X., Zhang H., Boyce B.F., Anolik J.H, & Xing L. (2018). B cells inhibit bone formation in rheumatoid arthritis by suppressing osteoblast differentiation. Nature Communications, 9, 5127.

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Histomorphometric Analysis of Vertebral Bone

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After micro-CT scanning, L3-L5 vertebrae were decalcified for 3 wk using 10% EDTA at 4°C, processed, and embedded in paraffin. 3-μm-thick sections at the center of the vertebrae with fewest trabeculae, selected from a series of continuously cut sections to ensure the measurement of all the sections was comparable, were blindly quantified for histomorphometric parameters, including the structural trabecular bone parameters, BV/TV (%), Tb.Th (μm), Tb.N (#/mm), Tb.Sp (μm), and OB surfaces on H&E-stained sections, and OC parameters on TRAP-stained sections using an OsteoMeasure Image Analysis System (Osteometrics, Decatur, GA) ^{50 (link),51 (link)} following the recommendations of the ASBMR Histomorphometry Nomenclature Committee ^{53 (link)}. T12 to L2 vertebrae were processed as LR white plastic-embedded blocks, as we reported previously ^{50 (link),51 (link)}. 3 μm-thick plastic sections were cut using a carbide steel knife on a Shandon Microtome. Sections at the center of the vertebrae were collected to evaluate the dynamic parameters of bone formation using an OsteoMeasure Image Analysis System, as we reported previously ^{50 (link),51 (link)}, following the recommendations of the ASBMR Histomorphometry Nomenclature Committee ^{53 (link)}.

Yao Z., Ayoub A., Srinivasan V., Wu J., Tang C., Duan R., Milosavljevic A., Ebetino F., Frontier A, & Boyce B. (2024). Hydroxychloroquine and a low activity bisphosphonate conjugate prevent and reverse ovariectomy-induced bone loss in mice through dual antiresorptive and anabolic effects. Research Square.

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Histomorphometric Analysis of Bone Mineralization

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At 4 days and 1 day prior to sacrifice, mice were injected subcutaneously with calcein (10 mg/kg) to label mineralizing bone. Distal femora were processed for static and dynamic histomorphometry as previously described. (18 (link)) Sections were stained according to the Von Kossa method with a tetrachrome counter stain (Polysciences, Warrington, PA) for assessment of cell-based measurements. Unstained sections were used for assessment of bone area and dynamic histomorphometry. Bone area/tissue area (%), trabecular number (mm^-1), trabecular thickness (μm), trabecular spacing (μm), mineralizing perimeter/bone perimeter (double-labeled perimeter + ½ single-labeled perimeter, %), mineral apposition rate (μm/day), bone formation rate/bone perimeter (μm²/μm/y), bone formation rate/bone area (%/year) and osteoclast perimeter/bone perimeter (%) were determined using the Osteomeasure image analysis system (OsteoMetrics Inc., Atlanta, GA) as described. (18 (link)) Additionally, mineralizing perimeter/bone perimeter (%), mineral apposition rate (μm/day) and bone formation rate/bone perimeter (μm²/μm/y) were determined on the endocortex of the femoral diaphysis. All data are reported using standardized units and nomenclature for bone histomorphometry. (27 (link))

Hawse J.R., Pitel K.S., Cicek M., Philbrick K.A., Gingery A., Peters K.D., Syed F.A., Ingle J.N., Suman V.J., Iwaniec U.T., Turner R.T., Spelsberg T.C, & Subramaniam M. (2014). TGFβ Inducible Early Gene-1 Plays an Important Role in Mediating Estrogen Signaling in the Skeleton. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 29(5), 1206-1216.

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