Hematology: Supravital Stains and Erythropoiesis

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Questions and Answers

Which of the following stains is NOT used for the demonstration of reticulocytes?

  • Brilliant Cresyl Blue
  • Eosin Y (correct)
  • Giemsa Stain (correct)
  • New Methylene Blue

What is the N:C ratio of a rubriblast?

  • 4:1 (correct)
  • 1:1
  • 3:1
  • 8:1

Which of the following statements best describes the cytoplasm of a rubriblast?

  • Small amount with dark blue hue due to ribosomes (correct)
  • Homogeneous with high eosinophilia
  • Bright pink due to hemoglobin concentration
  • Inhomogeneous with a dark blue color

Which feature does NOT characterize the maturation of erythroid precursors?

<p>Cytoplasm shifting from pink to blue (A)</p> Signup and view all the answers

Basophilia in cells is mainly attributed to which factor?

<p>High levels of ribosomal RNA (B)</p> Signup and view all the answers

What happens to the nuclear diameter during the maturation of reticulocyte precursors?

<p>Decreases significantly (D)</p> Signup and view all the answers

What is the visual assessment used to determine the N:C ratio?

<p>Area occupied by the nucleus compared to the cytoplasm (C)</p> Signup and view all the answers

Which term describes the pinkness of a cell due to the accumulation of basic components?

<p>Eosinophilia (A), Acidophilia (C)</p> Signup and view all the answers

Flashcards

Reticulocyte Staining

Reticulocytes are identified by stains like New Methylene Blue and Brilliant Cresyl Blue, which reveal the reticular structures in their cytoplasm.

Polychromatophilic Erythrocytes

Immature red blood cells, stained with a mixture of colors, appearing mixed or variable colors, indicating remaining RNA.

Diffuse Basophilic Erythrocytes

Immature red blood cells displaying a uniform, widespread blue color due to significant RNA.

N:C Ratio

The ratio of nucleus size to cell size, used to assess the maturation stage of blood cells.

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N:C Ratio (1:1)

Nucleus and cytoplasm are roughly equal in size.

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N:C Ratio (1:5)

Nucleus is smaller than the cytoplasm.

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N:C Ratio (3:1)

Nucleus is significantly larger than the cytoplasm.

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Basophilia

The blueness of a cell component, caused by attraction to basic stains like methylene blue (due to RNA).

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Eosinophilia

The pinkness of a cell component, caused by attraction to acidic stains like eosin (due to hemoglobin).

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Rubriblast (Pronormoblast)

The earliest recognizable red blood cell precursor, characterized by a large round nucleus and a small amount of dark blue cytoplasm.

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Rubriblast size

12-19 μm, in diameter

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Rubriblast nucleus shape

Round or slightly oval, central or slightly eccentric

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Rubriblast nucleus:cytoplasm ratio

About 4:1 (Turgeon)

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Study Notes

Supravital Stains for Reticulocytes

  • Supravital stains are used to identify reticulocytes in a Wright-stained smear.
  • New methylene blue and brilliant cresyl blue are examples of supravital stains.

Erythroid Precursor Maturation

  • As erythroid precursors mature, the overall and nuclear cell diameters decrease.
  • Chromatin condenses and clumps.
  • Nucleoli disappear.
  • Cytoplasm changes from blue to gray-blue then to salmon pink.

N:C Ratio

  • N:C ratio is a morphological feature to identify and stage RBC and WBC precursors.
  • It's a visual estimate of the nucleus area relative to the cytoplasm area.
  • If areas are equal N:C ratio is 1:1
  • If nucleus is less than 50% of the cell, N:C ratio is lower (e.g., 1:5).
  • If nucleus is more than 50% of the cell, N:C ratio is higher (e.g., 3:1).

Basophilia

  • Basophilia refers to the blueness of a particular part of the cell.
  • It's due to acidic components attracting basic stains like methylene blue.
  • Degree of cytoplasmic basophilia correlates with ribosomal RNA quantity.

Eosinophilia/Acidophilia

  • Eosinophilia/Acidophilia is the pinkness of a particular part of a cell.
  • It's linked to the accumulation of basic components that attract acid stains like eosin.
  • As RBC matures, eosinophilia correlates with hemoglobin accumulation, which decreases basophilia.

Rubriblast (Pronormoblast) Characteristics

  • Size: 12-19 μm
  • Nucleus: round to slightly oval, thin nuclear membrane, centrally or slightly eccentric.
  • Nucleoli: 1-2
  • Cytoplasm: small, dark blue, homogenous (due to ribosome concentration).
  • N:C ratio: 4:1 (Turgeon)
  • This cell (rubriblast) gives rise to prorubricytes.
  • Rodak (8:1): very far away, not yet wed.
  • Earliest recognizable precursor using light microscopy; not the youngest precursor.

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