Tag Archives: Granulocyte

Toxic Granules in Neutrophils

23 Mar IMG_1153 - Toxic Granules

IMG_1153 - Toxic GranulesGranulocytes have two types of granules primary and secondary. Primary granules are azurophilic, most numerous and prominent at the promyelocyte stage (see morphology of myeloid precursors) and diminish in number with further maturation. As the granulocyte matures the staining characters of the primary granules changes. They initially become violet and then became inapparent because they fail to take up stain. The secondary granules appear in the myelocytes and persist for the rest of the ice of the granulocyte. Neutrophils have fine pink secondary granules.

In conditions of intense stimulation of neutrophil maturation the primay granules may continue to take up stain in mature neutrophil because of a higher concentration of acid mucosubstances. These are called as toxic granules and the change called toxic change. The toxic granules are so called because they were first described in patients with gram negative sepsis and endotoxemia but may be found under conditions of intense stimulation of neutrophil production. The may be seen in

  1. Infection
  2. Inflammatory diseases
  3. Pregnancy
  4. Use of haemopoietic growth factors (G-CSF or GM-CSF)

Myelocyte and Neutrophils – Band and Segmented

18 Mar Myelo Band Neutro Final

Myelo Band Neutro Final

Neutrophilic Cells. The images shows myelocyte (1), band neutrophils (2), two lobed neutrophil (3) and neutrophils (4)

The image about shows morphology of cells of the neutrophilic series. The first cell to show commitment to a particular granulcoytic series (eosinophilic, basophilic or eosinophilic) is the promyelocyte. The promyelocytes of the three series can however only be differentiated by electron microscopy. The earliest cell showing features of neutrophilic differentiation on staining with Romanowsky staining is the myelocyte. Myelocytes may be neutrophilic, eosinophilic and basophilic. Myelocyte is a round cell with a round to oval nucleus that may be eccentrically placed. The cytoplasm shows two types of granules.

  1. Primary granules: The primary granules are azurophilic (reddish-purple or burgundy color) and are remnants of the granules of  the promyelocyte stage. The myelocyte does not synthesize primary granules. As the cell matures the number of primary granules declines and they disappear by the cells matures to a polymorphonuclear neutrophil.
  2. Secondary granules: The the size and staining of secondary granules are specific to the type of granulocytes. In neutrophils these granules are fine and pink staining

As the myelocyte matures the nucleus becomes more indented and finally becomes lobed.

The image above captures these changes

  1. Cells labels (1) are myelocytes. These show a pink cytoplasm with few primary (azurophilic) and many secondary (fine pink) granules and have a oval eccentrically placed nucleus with clumped chromatin without a nucleolus.
  2. The myelocyte develops and indentation of the nucleus and matures to a metamyelocyte. The metamyelocyte matures to a band neutrophil (cells labeled 2). which is called so because the nucleus is band shaped. With maturation the nucleus develops lobulation. The differentiation between a band neutrophil and a two lobed neutrophil is arbitrary and of little practical importance. A band cell becomes a two lobed neutrophil when its nucleus develops a constriction that is more than half to two third of the nuclear width (cell labeled 3).
  3. A neutrophil usually has 2-5 nuclear lobes (cells labeled 4)

Neutrophil and Lymphocyte

20 May 20110405_IMG_0051

Image

The image above shows a lymphocyte (left) and a polymprphonuclear neutrophil (right) in the peripheral blood. The lymphocyte is a cell about 10μ in size with a dense purple nucleus and agranular, slightly basophilic (blue) cytoplasm. About 10% of the lymphocytes are larger and have more abundant cytoplasm which shows granules (large granular lymphocytes). This is a small lymphocyte. A polymorphonuelear neutrophil is 12-15μ a cell with a segmented nucleus (not visible in this picture because the nucleus is folded on itself) that has clumped chromatin and an eosinophilic (pink) cytoplasm and pink granules. The contrast between nuclear shape, chromatin, cytoplasmic staining and granulation of the two cells is evident.

Morphology of Myeloid Precurssors

4 Jun 060411_0115_Morphologyo3.jpg

The earliest morphologically distinct myeloid cell is a myeloblast. Cell with myeloid commitment at stages of differentiation between the pleuripotant stem cell and myeloblasts have been identified but these lack morphological characters of a myeloid lineage. Myeloid cells share a common precursors with the cells of the monocyte-macrophage system in the form of CFU-GM. CFU-GM matures into CFU-G, a precursor for myeloid cells and CFU-M, a precursor for cells of the monocyte-macrophage series. There are five morphological distinct stages of myeloid precursors, myeloblast, promyelocyte, myelocyte, metamyelocyte and the band form. Only the first three are capable of cell division. Myeloid cells in the peripheral blood are classified into neutrophils, eosinophils and basophils depending on the staining characteristics of the cytoplasmic granules. Morphological evidence of commitment one of the myeloid lines is seen at the myelocyte stage. Electron microscopically commitment can be seen at the promyelocyte stage.

Figure 1. Myeloblast, Myelocyte and Metamyelocyte

Myeloblast (Figure 1, 3 and 4): Cells 10-20μ in size with a large slightly round nucleus with fine chromatin showing no clumping. Fine chromatin gives the nucleus a sieve-like or a finely granular appearance. The nucleus shows 2-5 pale sky-blue nucleoli. The nuclear membrane is exceedingly thin. Lymphoblasts nucleus may be differentiated from myeloblasts by the coarser chromatin, fewer nucleoli and the clumping of chromatin near the nuclear membranes. The cytoplasm of a myeloblast is basophilic but the basophilia is less marked than the lymphoblast or pronormoblast. Of the three types of leukaemic lymphoblasts the L1 (small cells with few nucleoli and a thin rim of cytoplasm) and L3 (strongly basophilic cytoplasm with prominent vacuolation) can be easily differentiated from myeloblasts morphologically. Differentiation of myeloblasts from L2 blasts on morphological grounds alone may not be possible. Perinuclear clearing is a feature of pronormoblasts and is not seen in the myeloblast. By definition myeloblasts have no granules but some classify cells with a few granules as myeloblasts particularly in the presence of abnormal myelopoiesis. In addition to granulation leukaemic myeloblasts may show nuclear furrows and Auer rods. Three stages in the maturation of myeloblasts are recognized

  1. Type I: No granules
  2. Type II: Fewer than 20 granules, Auer rods may be seen
  3. Type III: More than 20 granules seen without a Golgi apparatus

Very immature blasts lack morphological features of any lineage making it impossible to classify them as myeloblast, monoblast, lymphoblast or pronormoblast. Lineage identification in such cases may be aided by the company they keep, immunophenotyping and staining for peroxidase and esterases.

Figure 2. Promyelocytes

Promyelocyte (Figure 2): Slightly larger than a myeloblast (15-25μ), the promyelocyte is the largest cell of the myeloid series. Its nucleus is slightly indented, has a fine chromatin (though coarser than the myeloblast) and has nucleoli. Later stages show a slight chromatin condensation along the nuclear membrane. The cytoplasm is basophilic with prominent primary azurophilic granules. Endoplasmic reticulum is prominent and takes the form of dilated vesicles. Cells have a perinuclear Golgi apparatus that manifests as a perinuclear halo, but only in normal cells. Tough it is not possible to differentiate the promyelocytes of the three granulocytic (neutrophilic, basophilic, eosinophilic) series on light microscopy, electron microscopic difference exist. Leukaemic promyelocytes may be of the hypergranular or the hypogranular variety. The hypergranular variety has numerous small granules and Auer rods. Patients usually present with pancytopenia. Apart from having few or no granules in the cytoplasm the hypogranular variety may have a bilobed or folded nucleus.

Figure 3. Myelocytes – Neutrophilic, Eosinophilic and Basophilic

Myelocytes (Figure 1,3, 4 and 5): Lineage specific granules appear in the myelocyte stage. The cell is slightly smaller (10-20μ) than the promyelocyte and has an eccentric nucleus, round to oval, flattened on one side. Nucleoli are small and are only seen on EM, chromatin is coarse. Primary (azurophilic) granules persist but as no new granules are synthesized their number decreases with each division. The ratio of primary to secondary granules in a mature neutrophil is 2-3:1. Primary granules contain myeloperoxidase. The granulation of the endoplasmic reticulum is lesser than promyelocyte and as myelocytes mature the cytoplasmic basophilia decreases and finally disappears. The myelocyte may be neutrophilic, eosinophilic or basophilic. The granules of neutrophilic myelocytes are lilac, eosinophilic myelocytes are orange-red and basophilic myelocytes are purple. The eosinophilic myelocyte may contain purple staining granules. These are structurally different from granules of basophilic myelocytes. Myelocytes are the last stage to have the capacity to divide.

Granulocyte granules

Table 1. Charecteristics of granulocyte granules. The colour of the letters is the same as that of the granules and the backgroung the colour of cytoplasm of the cell the granules are most prominant in.

Figure 4. Myeloblast, Myelocyte and Eosinophil

Metamyelocyte (figure 1): Myelocytes matures to a metamyelocyte with appearance of nuclear indentation. Metamyelocyte can not divide. Unlike the transformation of a promyelocyte to a myelocyte that is accompanied by distinct morphological changes (disappearance of nucleoli, appearance of secondary granules) the most pronounced change that takes place when a myelocyte transforms into a metamyelocyte is indentation of the nucleus. Time-lapse photography has shown that the myelocyte nucleus can developed pronounced indentation and then revert to an oval shape. The distinction between the two stages is physiological. Metamyelocytes can not synthesize DNA, myelocytes can. Fortunately the distinction between a myelocyte and metamyelocyte is of little clinical importance and the ability of the cell to synthesize DNA is only assayed for research.

Band and Segmented Granulocytes (Figure 3): The indentation of the nucleus characteristic of the metamyelocyte increases and the nucleus becomes ribbon shaped. The nucleus of this cell has parallel borders for most of its length, like a band, and the cell is thus known as the band form. As the cell matures the nucleus becomes segmented. The segments are connected by thin strands of nuclear material. A constriction of more than half or two-third is accepted as segmentation by some. Neutrophils usually have 2-5 segments and eosinophils usually have 2 segments. Segmentations of the basophil nucleus are obscured by the intensely granular cytoplasm. The basophil nucleus ahs 2-3 segments, appearing in the contain few granules

Figure 5. Basophilic Myelocyte and Metamyelocyte

Morphological Characteristics of Myeloid Precursors

Cell Size Nucleus Cytoplasm
Myeloblast 12-20μ Round, fine chromatin, nucleoli Blue usually without granules. Some azurophilic granules may be present
Promyelocyte 15-25μ Slightly indented nucleus often eccentrically placed. Chromatin coarser than a myeloblast. Nucleoli seen Blue cytoplasm with numerous azurophilic granules
Myelocyte 10-20μ Round to oval, indented, chromatin more coarse than promyelocyte, no nucleoli Most immature forms have less basophilia than the promyeloblast. Basophilia disappeaqs as cell matures. Secondary granules appear.
Metamyelocyte 12-18μ Indented, coarse clumped chromatin Pink with secondary granules.
Band Form 10-12μ Band shaped. Borders are parallel for most of the length. May be folded Pink with secondary granules

Neutrophils Drumsticks and Sessile nodules

14 May

One X chromosome of women is inactivated. In nrutrophils this may appear in one of three forms. Drumsticks are nuclear appendages 1.5μ in diameter. They are seen in 0.5-2.6% of neutrophils. The inactivated X chromosome may also appear as sessile nodules or as a condensation under the nuclear membrane. The frequency of drumsticks increases with nuclear segmentation. They may be seen in eosinophils but are uncommon as eosinophils have fewer lobes. Racquet forms have a central clearing and should not be confused with drumsticks. They are not inactivations of X chromosomes. The X chromosome is only inactivated in an individual with more than one chromosome. Drumsticks are not seen in individuals having only one X chromosomes [males (XY), Turner's syndrome (XO) and testicular feminization (XY)]. Contrary to expectations individuals who are XXX rarely have cells with two drumsticks. They have an increased incidence of sessile nodule. XXX is also characterized by fewer neutrophil segments. Drumsticks in XXX are less common than normal women. The incidence of drumsticks in patients with Klienfelter’s Syndrome (XXY) is lesser than normal women. Shift to left, CML and Down’s syndrome is characterized by a decreased drumstick count. It returns to normal in CML following treatment. Drumsticks are more frequent in women with isochromosome of the long are of X. Patients with megaloblastic anaemia and congenital hypersegmentation have a higher frequency of drumsticks in the peripheral smear.

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