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Related Experiment Videos

A two step stain for normal and leukemic monocytes using two different dyes applied in sequence.

L Kass1

  • 1Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio.

Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission
|March 1, 1992
PubMed
Summary
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A new two-step staining method effectively identifies monocytes in blood and bone marrow. This technique uses C.I. basic blue 54 and C.I. basic blue 141 for distinct purple cytoplasm and red nuclei staining in monocytes.

Area of Science:

  • Hematology
  • Cell Biology
  • Histotechnology

Background:

  • Accurate identification of monocytes is crucial for diagnosing various hematological disorders.
  • Existing staining methods may lack specificity or require multiple steps.
  • A need exists for a simple, reliable staining technique for monocyte enumeration.

Purpose of the Study:

  • To develop and validate a novel two-step staining procedure for monocytes.
  • To differentiate monocytes from other blood and bone marrow cells using specific cytological features.
  • To provide a reproducible staining method for routine diagnostic use.

Main Methods:

  • A two-step staining procedure was employed using unfixed cells.
  • Cells were initially exposed to a methanolic solution of C.I. basic blue 54.

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  • Subsequently, an aqueous alkaline buffered solution of C.I. basic blue 141 was added for combined staining.

    • Main Results:

    • Monocytes exhibited intense deep purple staining of the cytoplasm.
    • Monocyte nuclei were frequently stained red.
    • Other normal and abnormal blood and bone marrow cells did not show similar specific staining patterns.
    • The staining procedure was completed within 10 minutes.

    Conclusions:

    • The developed two-step staining method provides specific and distinct staining for monocytes.
    • This technique offers a reliable and efficient way to identify monocytes in blood and bone marrow specimens.
    • The procedure has potential applications in hematological diagnostics and research.