Lab 3 WBC Flashcards
5 different factors causing infections
Parasitic Bacterial (septic) Rickettsial Fungal Viral
5 different factors causing inflammation
Infection
Endotoxin mediated (LPS, gram +)
Sterile necrosis (pancreatitis, tumor, trauma)
Chemical (inj, toxins, acids, alkalines)
Immune mediated (lupus)
Lab examinations to evalyate severity, locality and ethiology of inflammation
Hematology: CBC ESR Cytology Serum biochemistry Microbiology Parasitology Immunology Molecular biology Morphological pathology, histology
CBC
complete blood count
amount of each cell and hemoglobin
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
Serum biochemistry, 2 categories
Routine parameters (substrates, enzymes) Acute phase proteins
What blood coagulants are used for WBC counting in MAMMALS and how do they work
Na2EDTA
K2EDTA
NaCitrate
Chelates Calcium
What blood coagulants are used for bird, fish and reptile samples?
Calcium or Lithium heparin
What two machines are used to count WBCs
Bürker chamber
Hematology analyser
WBC counting using Bürker chamber
Hagedorn pipette Whole blood Türk solution 25 large squares Divide by 10 N = 10^9/l
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
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
Which hematology analyser type is the most accurate
Laser method
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
What dilutions are used by hematology analysers?
WBC 10x
RBC: 200x
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)
Neutrophil granulocyte cell line
Myeloblast Promyelocyte Myelocyte Metamyelocyte (jugend) Band forms (stab) Segmented forms
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)
- Marginal (blood vessels, inner surface)
- Maturation (bone marrow, under differentiation)
- 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
