White Cell Basics: Maturation

The mature forms of the myeloid series (neutrophils,

eosinophils, basophils), all have lobed (segmented) nuclei.
The degree of lobation increases as the cells mature.

John Santangelo
The earliest recognizable myeloid cell is the myeloblast (10-20um dia)
with a large round to oval nucleus. There is fine diffuse immature
chromatin (without clumping) and a prominent nucleolus.
The cytoplasm is basophilic without granules. Although you might see
a small golgi area adjacent to the nucleus, granules are not usually
visible by light microscopy. You should not see blast cells in the
peripheral blood.
The promyelocyte (10-20um) is slightly larger than a blast. Its nucleus,
although similar to a myeloblast shows slight chromatin condensation
and less prominent nucleoli.
The cytoplasm contains striking azurophilic granules or primary
granules. These granules contain myeloperoxidase, acid phosphatase,
and esterase enzymes. Normally no promyelocytes are seen in the
peripheral blood.
At the point in development when secondary granules can be
recognized, the cell becomes a myelocyte.
Myelocytes (10-18um) are slightly smaller than promyelocytes and have
eccentric round-oval nuclei, often flattened along one side.
The chromatin is fine, but shows evidence of condensation. Nucleoli
might be seen in early stages but not in the late myelocyte.
Primary azurophilic granules are still present, but secondary granules
predominate. Secondary granules (neut, eos, or baso) first appear
adjacent to the nucleus. In neutrophils this is the "dawn" of neutrophilia.
Myelocytes are not normally found in the peripheral blood.
Metamyelocytes (10-18um) are slightly smaller than myelocytes. They
have kidney shaped indented nuclei and relatively dense chromatin,
especially along the nuclear membrane.
The cytoplasm is faintly pink with almost no blue background. Numerous
secondary granules (neutro, eos, or baso) clearly outnumber primary
granules.
Zero to one percent of the peripheral blood white cells may be
metamyelocytes (juveniles).
Bands, slightly smaller than juveniles, are marked by a U-shaped or
deeply indented nucleus.
Opposite sides or lobes are of roughly equal size or diameter. There
is no nuclear constriction > than 1/2 the lobe diameter.
The chromatin is heavily clumped and secondary or specific
granules either neutrophilic or basophilic predominate.
Normal band counts vary but are usually in the range of 0-6%.
Segmented (segs) or polymorphonuclear (PMN) leukocytes (average 14um
dia) are distinguished by definite lobation with thin thread-like filaments of
chromatin joining the 2-5 lobes. The chromatin of the segmented neutrophil
is coarsely clumped and the cytoplasm is pink due to large numbers of
secondary granules.
In practice when examining peripheral blood, neutrophils are the only
leukocytes to be divided into myelocyte, juvenile, band, and PMN stages.
Eosinophils and basophils of all stages are lumped together in most
instances.
Normally approximately 45-75% of the peripheral blood white cells are
segmented neutrophils.
 Myeloblast Promyelocyte Myelocyte

Metamyelocyte Band Segmented

 RBC Basics: Morphology
• Although most erythroid and myeloid cells
decrease in size as they mature, the
cytoplasm becomes relatively increased in
volume, compared to the nucleus.

• The cytoplasm becomes less basophilic and

increasingly eosinophilic.
 Erythroblast or pronormoblasts
• The pronormoblast (E1) is the earilest
recognizable red cell precursor.
• They are 15- 20µ in diameter with fine
granular chromatin and 0-2 nucleoli. The
cytoplasm is deeply basophilic without
granules.
 Basophilic normoblasts
• The chromatin is slightly coarser and the cell
slightly smaller than the pronormoblast.
Nucleoli may be indistinct.
• The cytoplasm remains deeply basophilic.
 Basophilic normoblasts
• The chromatin becomes increasingly coarse
compared to the E2 stage.
• The cytoplasm remains basophilic, but has a
lighter tinctorial quality than E2.
 Polychromatophilic normoblasts
• Hemoglobin production becomes evident from the muddy
mix of the blue staining RNA and the red staining
hemoglobin resulting in the greyish cytoplasm of the E4
polychromatophilic normoblast.
• The chromatin is moderately condensed.
 Orthochromatic normoblasts
• The E5 orthochromatic normoblast is smaller than
the E4 stage.
• In the E5 orthochromatic normoblast hemoglobin is
abundant while ribosomes decrease in number.
• The nuclear chromatin is pyknotic and ready to be
extruded.
 Polychromasia
• Polychromatophilic red cells are larger than mature
erythrocytes and still contain ribosomes and
mitochondria, giving the cell a blue tint.
• When polychromatic RBCs are stained with a
supravital dye (new methylene blue or cresyl violet)
ribosomes clump to form a blue stained granulars
called reticulum. These are called reticulocytes.
 Mature red cells
• Normal mature erythrocytes are biconcave discs
(approximately 7 um in diameter; 2.5 um thick at the
periphery; 1.0 um thick at the center).
• This results in an area of central pallor (about 1/3
the dia.) seen in mature RBCs stained with Wright's-
Giemsa.

Erythrocytes
 Megakaryocytes
• Megakaryocytes undergo endomitosis - a process
whereby DNA is duplicated without cell division.
• Megakaryocytes are the largest normally occurring
cells in the marrow (35-160um)

Platelet
Precursors
A Cyte for sore eyes