Lab 3 WBC Flashcards by Lea Damler

5 different factors causing infections

Parasitic
Bacterial (septic)
Rickettsial
Fungal
Viral

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5 different factors causing inflammation

Infection
Endotoxin mediated (LPS, gram +)
Sterile necrosis (pancreatitis, tumor, trauma)
Chemical (inj, toxins, acids, alkalines)
Immune mediated (lupus)

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Lab examinations to evalyate severity, locality and ethiology of inflammation

Hematology:
CBC
ESR
Cytology
Serum biochemistry
Microbiology
Parasitology
Immunology
Molecular biology
Morphological pathology, histology

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CBC

complete blood count

amount of each cell and hemoglobin

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ESR

erythrocyte sedimentation rate
amount of plasma on top of AC blood after 1 hr
Higher amount of protein (CRP) makes the RBC sediment quicker

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Serum biochemistry, 2 categories

Routine parameters (substrates, enzymes)
Acute phase proteins

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What blood coagulants are used for WBC counting in MAMMALS and how do they work

Na2EDTA
K2EDTA
NaCitrate
Chelates Calcium

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What blood coagulants are used for bird, fish and reptile samples?

Calcium or Lithium heparin

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What two machines are used to count WBCs

Bürker chamber

Hematology analyser

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WBC counting using Bürker chamber

Hagedorn pipette
Whole blood
Türk solution
25 large squares
Divide by 10
N = 10^9/l

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What is Türk solution

Hemological stain
Crystal violet or liquid methylene blue in acetic acid

Hemolyses erythrocytes and membrane of all cells, but stains the nuclei of WBC, making them well visible and easy to count

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WBC counting by hematology analyser, 4 different ways

Light absorbancy of different cell layers (more accurate)
Laser technique (scattering)
Reagents to measure peroxidase activity of the phagocytic cells
Flow cytometric method

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Which hematology analyser type is the most accurate

Laser method

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How do automatic cell counters work?

They make a distribution according to the impedance of a particle entering between two electrodes
First they dilute the sample, then hemolyse the RBC.
They differentiate between the sizes of the particles. Reagents are used to shrink the lymphocytes and increase the size of neutrophils and macrophages

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What dilutions are used by hematology analysers?

WBC 10x

RBC: 200x

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Laser cell counters

Laser light through cells: millions of scatters are detected by two light detectors:
1. Low angular/forward: size
2. High angular/side: inner structure
Quantity of light energy (lux)

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Neutrophil granulocyte cell line

Myeloblast
Promyelocyte
Myelocyte
Metamyelocyte (jugend)
Band forms (stab)
Segmented forms

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Neutrophil function and %

First cells to reach site of injury, lines endothel
Phagocytic function
Segmented form: 60-77%, 3-11.8 x 10^9

Lymphoid cell line

Lymphoblast
Small lymphocyte
Middle sized lymphocyte
Reactive T lymphocyte

Lymphocyte function and %

Humoral immune system, resides in lymph tissues, B-cells produce antibodies
12-30%, 1-4,8 x 10^9

Eosinophil cell line

Young form (band nucleated)
Segmented form

Eosinophil function and %

Granules with toxins to kill pathogens, parasites and worms, play a role in allergy
2-10%, 0.1-1.35 x 10^9

Basophil cell line

Young form (band nucleated)
Segmented form

Basophil function and %

Anticoagulants, antibodies, Histamine producing
Dilates blood vessels
<1%

Monocyte cell line

Monocytic (young) form | Reactive macrophage

Monocyte function and %

Phagocytosis of all dead cells | 1-5%, 0.1-0.5 x 10^9

WBC pools in the body

Bone marrow: mitotic, maturation, storage Blood vessel: marginal pool (high mobilisation) Tissue pool

Physiological leukocytosis

Acute/chronic stress (EP, NE, GC)

Acute inflammation | First lab sign

Increase in positive acute phase proteins | Decrease negative acute phase proteins

Neutropenia

Migrating factors produced by tissue cells: macrophages. Neutrophils migrate from blood to site of inflammation, which leads to a total decrease or WBC count

Neutrophilia

G-CSF and GM-CSF increases WBC production in the bone marrow. The factors are produced by macrophages

Left shift

Many young WBCs | Metamyelocytes (jugend) and band (stab) forms

Where are WBCs mobilized from (in what order)

1. Marginal (blood vessels, inner surface) 2. Maturation (bone marrow, under differentiation) 3. Mitotic (bm, very young c lols, proliferating)

Regenerative left shift, prognosis

Inc WBC count, neutrophilia, left shift Bone marrow regenerates new cells Favorable prognosis

Degenerative left shift, prognosis

Low or normal WBC count, neutrophils immature (Neutropenia) Bone marrow cannot keep up Severe bacterial inflammation Poor prognosis

Leukemoid reaction

Due to increased G-CSF and GM-CSF, the WBC count can be very high (>70) in peripheral blood Typical: big abscesses, endometritis (pyometra) Blood smear: can be confused with chronic myelogenous leukemia

Toxic neutrophils

Granulocytopoiesis disturbed in severe inflammatory reactions Azurophilic granules produced in basophilic cytoplasm

Döhle-bodies

Angular basophilic inclusion bodies in neutrophil granulocytes (remnants of ER)

Right shift

Many segmented and hypersegmented neutrophils (old) | Reduced amount of young, newly produced

Name three acute phase proteins (APP)

CRP Fibrinogen Haptoglobin

What species are lymphophilic?

Swine, sheep, poultry

What species are neutrophilic?

Dog, cat, horse

What is leukemia?

Cancer of the body’s blood forming tissues | Lymph system, bone marrow etc

What is the difference between positive and negative APP?

Positive: plasma conc increases during infl Negative: plasma conc decreases during infl