Calcium Salt Stain

Calcium Salt Stain (Sliver nitrate staining method)


In the field of histology, accurate staining techniques are essential for identifying and studying tissue structures. One such method, the Metal Replacement Method, offers a reliable way to highlight specific elements within histological sections. This article delves into the main components, test procedure, result interpretation, and precautions associated with the Metal Replacement Method. If you’re a professional in the field of pathology or simply curious about this staining technique, read on to explore its intricacies and importance.

Main Components of Calcium Salt Stain

The Metal Substitution Strategy includes the utilization of explicit reagents to accomplish the ideal outcomes. The fundamental parts include:

  • Silver Nitrate Arrangement: This contains silver nitrate as the essential fixing.
  • Sodium Thiosulfate Arrangement: This contains sodium thiosulfate as the essential fixing.

Test Procedure

To achieve successful staining with the Metal Replacement Method, follow these steps:

  • Dewaxing and Washing: Regularly dewax the tissue segments and wash them with refined water.
  • Silver Nitrate Treatment: Drop silver nitrate arrangement on the tissue segments or drench them in the arrangement. Open them to solid daylight for 15 to an hour, or utilize bright (UV) light for 5 to 10 minutes. Then, flush the areas with refined water for 3 minutes.
  • Sodium Thiosulfate Treatment: Drench the segments in sodium thiosulfate answer for 2 minutes, trailed by washing with running water for 5 minutes.
  • Hematoxylin Staining: Spot the segments in Hematoxylin (Harris) for 4 to 6 minutes, and marginally wash them with water.
  • Separation: Separate the segments in 1% hydrochloride liquor answer for a couple of moments, and flush with running water.
  • Eosin Staining: Stain the segments in Eosin stain (Fluid) briefly, and marginally flush with water.
  • Dehydration and Sealing: Conventional dehydration and transparent process, followed by sealing with neutral resin.

Result Interpretation

The Metal Replacement Method produces distinctive staining outcomes. Calcium salt Stain within the histological section appears brown-black to deep black after the process is completed.


To ensure accurate and safe usage of the Metal Replacement Method, adhere to the following precautions:

  • Use Neutral Buffered Formaldehyde Solution: Always fix calcium salt with neutral buffered formaldehyde solution instead of an acidic fixative to prevent the dissolution of a portion of the salt.
  • Consider Counterstaining: If needed, consider counterstaining the sections with Van Gieson solution instead of H-E solution.
  • Professional Use and Interpretation: This staining reagent should be handled and interpreted by trained professionals. Carefully read the instruction manual before use, adhere to the validity period, and implement personal hygiene protection.
  • Proper Waste Disposal: Dispose of waste following the guidelines of the hospital or environmental protection department.
  • Check Packaging Information: Ensure verify the production date, production batch number, and expiration date on the outer packaging.


  • Chinese Medical Association. Clinical Technical Operating Specifications – Pathology Volumes [M]. People’s Military Medical Publishing House.
  • Ling Qibo. Practical Pathology Special Staining and Histochemistry Technology [M]. Guangdong Higher Education Press.

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