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Histology Winter Semester > Peripheral Blood and Hematopoiesis (Lecture 10 and 11) > Flashcards

Peripheral Blood and Hematopoiesis (Lecture 10 and 11) Flashcards

1
Q

Explain the process of preparation, staining and evaluation fo the blood smear

A

Blood cells are studied in smears prepared by spreading of a drop of periferal blood in a thin layer on the microscope slide.

Procedure:

  1. Disinfection of the skin of the fingertip of the laft fourth or thrid finger (on right handers)
  2. Puncture into the ball of the fingertip with sterile lancet or single use needle
  3. The first drop of blood is wiped off because blood is diluted by tissue fluid
  4. The second drop of blood is placed near an end of a glass (microscope) slide and spread using another slide (spreader slide)
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2
Q

Explain the process of spreading

A

Spreading:

  • Spreader slide is moved over the glass slide at an angle of 45 degrees, when slide edge touches the blood drip, the blood spreads by capillary forces along its edge.
  • A thin film of blood is obtained by a smooth quick motion fo the spreader slide acrosss a glass slide.
  • The air dried blood smear is flixed and stained.
  • Blood smears are stained with mixutres of acidic (eosin) and basic dyes (thethylene blue and its oxidative products - methylene violet and azure)
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3
Q

What is Pappenheim’s method?

A

Method is used for staining of peripheral blood and bone marrow smears.

  1. Fixation of blood smear with May-Grünwald solution
    (eosinate of methylene blue in methanol)
    3 minutes
  2. Staining with diluted May-Grünwald solution
    (the same amount of distilled water was added)
    1 – 2 minutes
  3. Pour off mixture
  4. Staining with Giemsa-Romanowsky
    (eosinate of the methylene azure, blue and violet)
    15 minutes
  5. Washing (distilled water)
  6. Air-drying

Results of staining:

Red blood cells – pink/red

Nuclei – purple/blue

Neutrophilic granules – salmon pink

Eosinophilic granules – brick-red

Basophilic granules – blue/violet

Cytoplasm of lymphocyte – sky blue

Cytoplasm of monocyte – pale blue (grayish or greenish)

Azurophilic granules - purple red

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4
Q

How are the blood smears evaluated?

A

Blood smear is observed in light microscope using HI objective (oil.im.,100x) and immersion oil.

RBC evaluation:

  • Size, shape, structure
  • Anisocytosis (microcytosis, macrocytosis)
  • Poikilocytosis - variation in RBC shape:
    • Spherocytosis, ovalocytosis, sickle cells

WBC evaluation:

  • Size and morphology
  • Leukogram (differential leukocyte count)
    • Proportional incidence (%)

Arneth formula:

  • Reflects ratio of the neutrophilic granulocytes based on the number of nuclear lobes.
  • Shift to left - predominance of the young forms (band and two lobes).
  • Shift to right - predominance of the old forms (four and five lobes).
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5
Q

What are the formed elements in the blood cell count?

A

ERYTHROCYTES:

  • Male: 4.5-6.0 x 1012 /L
  • Female: 4.0-5.2 x 1012 /L

LEUKOCYTES:

  • 4.000 - 10.000 /μL
  • 4 – 10 x 109 /L

LEUKOGRAM (differential leukocyte count):

  • Neutrophils: 60-70 % (band form: 2-5 %)
  • Eosinophils: 2-5 %
  • Basophils: 0-1 %
  • Lymphocytes: 20-40 %
  • Monocytes: 2-10 %

THROMBOCYTES:

  • 150,000 - 400,000 /μL
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6
Q

What is this slide?

A

Blood smear with lymphocytes and neutrophilic granulocyte

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7
Q

What is this slide?

A

Band form of a neutrophil and neutrophilic granulocyte

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8
Q

What is this slide?

A

Lymphocyte and an eosinophil

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9
Q

What is this slide?

A

Monocyte

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10
Q

What is this slide?

A

A neutrophilic granulocyte and basophilic granulocyte and a monocyte

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11
Q

What are the periods of hematopoiesis?

A
  • Blood islets - yolk sac mesoderm – since 3rd week
  • Liver, spleen - starts at the 2nd month
  • Bone marrow – since the 3rd month
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12
Q

What is bone marrow?

A

Red bone marrow and yellow bone marrow

Red (hematogenous) bone marrow:

  • Stroma, hematopoietic cords and sinusoidal capillaries
    • Stroma – reticular connective tissue - reticular cells, reticular fibers and macrophages
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13
Q

What is hematopoiesis?

A
  • Pluripotent and multipotent stem cells
  • Progenitor and precursor cells (blasts)
  • Self renewal, proliferative activity
  • Myeloid and lymphoid multipotential cells (CFU-S and CFU-L)
    • CFU = colony forming units
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14
Q

How can the bone marrow be examined?

A

Sternal puncture

OR

Trepanobiopsy - bone marrow + bone tissue

  • A sample from the iliac bone is taken and is processed using histological techniques
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15
Q

What is this slide?

A

Hematopoietic tissue from red bone marrow

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16
Q

What is the process of erythropoiesis?

A
  1. Proerythroblast – around 20 μm, large spherical nucleus, nucleoli, basophilic cytoplasm
  2. Basophilic erythroblast – around 10-16 μm, spherical nucleus, nucleoli not visible, basophilic cytoplasm
  3. Polychromatophilic erythroblast – around 10-12 μm, smaller nucleus – condensed chromatin (clock face), baso- and eosinophilia in the cytoplasm
  4. Orthochromatophilic erythroblast – around 8-10 μm, small nucleus with highly condensed chromatin, nucleus is extruded at the end, eosinophilic cytoplasm
  5. Reticulocyte

Changes that occur during maturation:

  • Cell volume decreases
  • Cell nucleus gets smaller
  • Basophilia → eosinophilia
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17
Q

What is this slide?

A

Proerythroblast

18
Q

What is this slide?

A

Proerythroblast and plasma cell

19
Q

What is this slide?

A

Basophilic erythroblasts

20
Q

What is this card?

A

Polychromatophilic erythroblast

21
Q

What is this slide?

A

Orthochromatophilic erythroblast

22
Q

What is this slide?

A

Orthochromatophilic erythroblast

23
Q

What is this slide?

A

From left to right:

Basophilic erythroblast

Polychromatophilic erythroblast

Orthochromatophilic erythroblast

24
Q

What is granulopoiesis?

A

Myoblast –> 15-20um, spherical nucleus, nucleoli

Promyelocyte I –> 22-25um, azurophilic granules

Promyelocyte II –> First specific granules from (neutrophilic, eosiophilic, basophilic)

Myelocyte –> 15-20um, oval nucleus, coarser chromatin clumps

Metamyelocyte –> Horse-shoe nuceus, postmitotic

25
Q

What is this slide?

A

Myeloblast

26
Q

What is this slide?

A

Myeloblast

27
Q

What is this slide?

A

Promyeloblast

28
Q

What is this slide?

A

Promyelocyte

29
Q

What is this slide?

A

Neutrophilic myelocyte

30
Q

What is this slide?

A

Eosinophilic myelocyte

31
Q

What is this slide?

A

Neutrophilic metamyalocyte

32
Q

What is this slide?

A

Eosinophilic metamyelocyte

33
Q

What is this slide?

A

Basophilic metamyelocyte

34
Q

What is monocytopoiesis?

A

Monoblast

Promonocyte

Monocyte

35
Q

What is this slide?

A

Promonocyte

36
Q

What is lymphopoiesis?

A

Lymphoblast

Prolymphocyte

37
Q

What is thrombocytopoiesis (megakaryocytopoiesis)

A

Megakaryoblast

Promegakaryocyte

Megakaryocyte

38
Q

What is this slide?

A

Megakaryoblast

39
Q

What is this slide?

A

Promagakaryocyte

40
Q

What is this slide?

A

Megakaryocyte

Histology Winter Semester (10 decks)

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