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Special Stains

Several special stains are performed by this laboratory. Please contact the lab if stains other than those listed here are required. Consultation is available on selection of appropriate staining protocols for research projects.

  • H&E: Routine stain for cellular detail, best on paraffin sections
  • Toluidine Blue O: Stain utilized for optimal demonstration of mineralized bone and osteoid seams, osteoblasts, osteoclasts.
  • Alkaline Phosphatase: Enzyme histochemical stain for osteoblasts. This stain is diminished in formalin fixed tissues.
  • Tartrate Resistant Acid Phosphatase (TRAP): Enzyme histochemical stain specific for osteoclasts.
  • Von Kossa: Stain best for demonstration of mineralization in bone and other tissues. Also used to demonstrate areas of mineralization in cell cultures.
  • Safranin O: Stain which demonstrates presence of proteoglycans.
  • Gomori’s trichrome: Stain used to visualize collagen in tissues
  • EVG (Elastin/Van Gieson: Demonstrates presence of elastic fibers in tissues
  • Prussian Blue: Demonstrates presence of iron in tissues
  • Goldners Trichrome: Stain utilized for differentiation of mineralized and non-mineralized areas in bone.
  • Lee’s Methylene Blue: H&E “lookalike” stain used for GMA sections.
  • Alcian Blue: Stains acid mucins in tissues.
  • Alizarin red/Alcian blue staining of whole tissues: Stain for differentiation of mineralized vs. unmineralized bone in whole tissues
  • In vivo fluorescent bone labeling: Calcein, xylenol orange and alizarin red labeling in vivo for bone mineralization front.
  • Calcein: In vivo labeling of bone mineralization fronts suitable for all species. Preparation of Calcein for injection is available upon request.
  • Oil Red O (frozen sections only): Stain which demonstrates the present of lipid droplets in tissues. Suitable only for tissues that have been sectioned frozen and not processed through any solvents.
  • Osmium Staining: Marrow adipose can be quantified by using this heavy metal to infuse lipid in situ. Lipid can then be visualized and quantified using μCT. This process uses the entire bone and requires decalcification.
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    Special stains are prepared on site to suit individual researcher’s needs.

    Photo by Robert A. Lisak
  • Photo

    Tartrate Resistant Acid Phosphatase
    425X magnification showing areas of tartrate resistant acid phosphatase activity in proximal tibia of the mouse. Osteoclasts are the cells responsible for the resorption of bone.

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    Safranin O
    250X magnification of mouse knee joint stained with Safranin O (red) demonstrating the distribution of proteoglycans in articular cartilage. Undemineralized mouse knee was embedded in methylmethacrylate and sectioned to a thickness of 5μm prior to staining.

  • Photo

    In vivo fluorescent bone labeling
    425 X magnification demonstrating calcein pulse-labeling of mineralization fronts in the trabecular bone of the proximal tibia of the mouse. Pulse labeling bone mineralization fronts allows quantitative measurement of mineralization rates.