Bone Decalcification Services 

Trusted Solutions for Preparing Hard Tissue for Histology, IHC, and Molecular Studies

🔬 Introduction 

Decalcification of bone tissue is a crucial step in the preparation of skeletal samples for histological examination. This process removes calcium ions from mineralized tissues, softening them sufficiently to allow thin sectioning on a microtome without damaging blades or compromising structural integrity.

The choice of decalcification agent can significantly affect:

• Tissue morphology and cellular detail

• Compatibility with staining techniques (H&E, IHC, ISH)

• Integrity of DNA, RNA, and protein targets

At iHisto, we offer both rapid acid-based and gentle chelating decalcification methods — tailored to your sample type, timeline, and downstream goals.

⚗️ Common Decalcification Methods

EDTA (iHisto Recommended)

Chelating method | Best for IHC & molecular studies

• Description: Ethylenediaminetetraacetic acid (EDTA) binds calcium ions at physiological pH (7.0–8.0) without damaging cellular components.

• Advantages:
  Best preservation of tissue architecture
  Compatible with immunohistochemistry, RNA in situ hybridization, and enzyme histochemistry
  Minimal tissue shrinkage or epitope loss

• Disadvantages:
  Slow: May require 1–3 weeks for large or dense bones

🔹 Recommended by iHisto for all sensitive downstream assays.

Formic Acid

Moderate-speed acid method | General-purpose use

• Description: A mild acid that removes calcium by both chelation and acid reaction.

• Advantages:
  Suitable for both small and large samples
  Compatible with most routine stains
  Faster than EDTA (1–4 days for small bones)

• Disadvantages:
  May cause mild tissue shrinkage
  Some IHC markers may be affected

Nitric Acid

Rapid acid method | For urgent turnaround

• Description: A strong mineral acid used when speed is prioritized over detail.

• Advantages:
  Extremely fast-acting (hours to 1 day)
  Useful for clinical or urgent preclinical samples

• Disadvantages:
  Poor preservation of fine histology
  May damage nuclei, membranes, and antigens

Hydrochloric Acid (HCl)

Strong acid method | Limited use

• Description: Another rapid decalcifier, used in some commercial solutions.

• Advantages:
  Effective for dense bones when time is limited

• Disadvantages:
  Can cause severe distortion, especially in fragile or highly cellular tissue
  Not recommended for IHC or ISH

Citric Acid

Mild organic acid | Niche applications

• Description: A weak acid sometimes used for gentle decalcification.

• Advantages:
  Gentle on tissue, with better morphology than strong acids

• Disadvantages:
  Slower than formic or nitric acid
  Not commonly used for standard protocols

🧪 iHisto’s Decalcification Services Include:

• Sample trimming and cassette transfer

• Controlled decalcification timing with endpoint testing

• EDTA, formic, nitric, HCl, or custom buffer options

• Compatible with human, rodent, NHP, and orthopedic implant samples

• FFPE embedding or OCT cryo-embedding post-decalcification

• Full downstream workflow: sectioning, staining, TRAP, IHC, and digital imaging

✅ Why Choose iHisto

• 🧬 EDTA-based protocols for biomarker preservation

• 🔬 Custom buffer selection based on study goals

• 🧠 Experienced with bone, joint, spinal, and implant specimens

• 📷 Digital slide scanning and optional pathology review

• 🧊 Cold-chain sample intake and biobank compatibility

❓ FAQs

• How long does decalcification take?
  Depends on bone size and agent: EDTA (1–3 weeks), formic acid (1–4 days), nitric acid (4–24 hours).

• Do you provide sectioning and staining after decalcification?
  Yes — we offer H&E, trichrome, Safranin O, TRAP, IHC, and digital slide services.

• Can you process implant-associated or metal-adjacent bone?
  Yes. We support evaluation around orthopedic hardware and biomaterials.

• Can I choose the decalcification method?
  Absolutely. Let us know your priority (speed vs. preservation), and we’ll recommend the best option.

📩 Request a Quote or Consultation 

Whether you're working with rodent femurs, spinal bone, or human biopsies, iHisto delivers precise, customized decalcification workflows tailored to your research or diagnostic needs.

👉 Contact us or email info@ihisto.io to get started.