Reticulocyte Stain

• Specifications:6x20mL, 2x100mL (Box)
• Expected use: Planned for staining of reticulocyte and hemoglobin H consideration body in entire blood
• Store temperature: 5℃～30℃

Introduction

Reticulocyte Stain serves as a critical indicator of red blood cell health and production within the body. Positioned between the stages of young, developing red blood cells and fully mature ones, reticulocytes offer valuable insights into the body’s hematopoietic activity. Reticulocyte staining, a specialized laboratory technique, facilitates the visualization and quantification of these transitional cells. In this article, we will delve into the fundamental principles, procedural steps, and the importance of reticulocyte staining in the realm of hematology.

Principle of Reticulocyte Stain

The underlying principle of reticulocyte staining lies in the unique composition of these cells. Reticulocytes contain basophilic RNA within their cytoplasm, a characteristic absent in fully mature red blood cells. By employing a specially formulated reticulocyte staining solution, these cells become distinguishable through microscopic examination. The staining solution generates a distinct mesh-like pattern that can appear as either light blue or dark blue, enabling easy identification of reticulocytes.

Components of the Staining Solution

The reticulocyte staining solution predominantly incorporates a vital component known as “New Methylene Blue.” This element interacts with the basophilic RNA present in the reticulocytes’ cytoplasm, leading to the formation of the characteristic mesh structure visible upon microscopic analysis.

Sample Requirement and Test Procedure

For accurate outcomes, it is imperative to utilize either fresh whole blood or blood treated with EDTA·K2 anticoagulant. The staining process entails two key procedures: reticulocyte staining and hemoglobin H inclusion body staining.

Reticulocyte Staining Procedure:

• Combine the reticulocyte staining solution and whole blood in a 1:1 ratio.
• Allow the mixture to stand at room temperature for a minimum of 20 minutes.
• Create a blood smear from the mixture, followed by allowing it to dry. Subsequently, observe the slide under a microscope for result interpretation.

Hemoglobin H Inclusion Body Staining Procedure:

• Mix the reticulocyte staining solution and whole blood in a 2:1 ratio.
• Introduce a plug to the mixture and place it within a 37℃ water bath for a duration of 4 to 18 hours.
• Prepare a blood smear from the mixture, permit it to dry, and then examine it under a microscope to interpret the findings.

Result Interpretation

• Reticulocyte Staining: Reticulocytes display a cytoplasm featuring a mesh structure in shades of light blue or dark blue. By analyzing 1000 red blood cells using an oil microscope, the percentage of reticulocytes can be calculated.
• Hemoglobin H Inclusion Body Staining: Circular dark green and blue structures of varying sizes become visible within red blood cells. These structures, referred to as hemoglobin H inclusion bodies, are irregularly distributed within the red blood cells. Similar to reticulocyte staining, the percentage of red blood cells positive for HbH inclusion bodies can be determined by analyzing 1000 red blood cells under an oil microscope.

Precautions and References

Several precautions need to be followed during the staining process to ensure precise outcomes and adhere to safety protocols. Moreover, this technique finds mention in established clinical laboratory manuals, such as “National Clinical Laboratory Procedures” by the State Health and Family Planning Commission of the People’s Republic of China Medical Administration Bureau and “Gradwohl’s Clinical Laboratory Methods And Diagnosis, 8th ed.”

Conclusion

Reticulocyte staining plays a pivotal role in understanding red blood cell composition and health, aiding in the diagnosis and monitoring of various blood disorders.

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