agile - affordable - accurate Basics of Hematology cell counting

agile - affordable - accurate BASICS OF HEMATOLOGY Functions of blood Transport through the body of –oxygen and carbon dioxide –nutrient molecules (glucose, lipids, amino acids) –ions (e.g., Na +, Ca 2+, HCO 3− ) –„wastes” and metabolites (e.g., urea) –hormones –Heat Defense of the body against infections and other foreign materials. All the WBCs participate in these defenses.

agile - affordable - accurate Composition of the Blood LIQUIDPlasma CELLS Erythrocytes (Red Blood Cells) 4-6 *10 12 /l Leukocytes (White Blood Cells) 6-15 *10 9 /l Thrombocytes (platelets = cell parts) 0,2-0,8 *10 12 /l Proteins (7%)Hormones LipoproteinsAlbuminGlobulinsFibrinogen Nutrients, metabolits, bioactive molecules (<1%) Free Amino acids Free Amino acidsVitaminsGlucoselipoids Salts (1%) Water (91-92%)

agile - affordable - accurate Noncellular components Plasma is the straw-colored liquid in which the blood cells are suspended. Contains coagulation factors. After coagulation it is called serum. Composition of blood plasma –various ions (Na +, Ca 2+, HCO 3−, etc.) –proteins (albumins, globulins) –glucose and traces of other sugars –amino acids –other organic acids –cholesterol and other lipids –hormones –urea and other wastes

agile - affordable - accurate Cellular components of blood 1 Blood is a liquid tissue. Suspended in the watery Plasma are seven types of cells and cell fragments. (They can be counted in particle counters.) Red blood cells (RBCs) or Erythrocytes Platelets or thrombocytes (PLT) five kinds of white blood cells (WBCs) or Leukocytes Neutrophils Lymphocytes Eosinophils Basophils Monocytes 3-part counters: RBC, PLT and neutrophils lymphocytes, eosinophiles+basophiles+monocytes

agile - affordable - accurate Cellular components of blood 2

agile - affordable - accurate Main Cellular Parameters All of these parameters strongly depend on properties of the surrounding medium and varies by species! Morphology: can be visualized in smears by different staining methods (microscopy) Cell numbers Mean Corpuscular Volume (MCV) of RBC’s Packed cell volume of RBC’s

agile - affordable - accurate Red Blood Cells (RBC) RBCs form the majority of the blood cells Non nucleated in mammals Biconcave shape Structures on the cell membrane determine blood group systems Contain hemoglobin Hemolysis: rupture of cell membranes followed by destroying the cells Hemolytic agents: detergents, pH, osmotic pressure, drugs, immunsystem etc. Reticulocytes (immature red blood cell forms) in a very few %

agile - affordable - accurate Function of Hemoglobin It causes the red color of blood has 4 subunits, contains iron Main function: oxygen transport MCH: Mean Corpuscular Hemoglobin (pg) MCHC: Mean Corpuscular Hemoglobin Concentration (g/l)

agile - affordable - accurate Hematocrit-HCT or Packed Cell Volume-PCV Volume of blood in % occupied by cells. It can be measured after separation in a centrifuge, or calculated after cell counting. Shows the degree of anemia. 100% 40-50% RBCs WBCs Plasma

agile - affordable - accurate Thrombocytes (= Platelets) Thrombocytes play an important role in the cellular part of the hemostasis (in plug forming) Non nucleated cells in mammals Contain specific granules Avian thrombocytes (nucleated) Mammalian thrombocytes

agile - affordable - accurate White Blood Cells (WBC) 1 - Granulocytes Neutrophil Neutrophils Neutrophils: Granules don't stain well with either acidic or basic dyes, therefore, neutral. The most frequent granulocyte. Eosinophils: Eosinophils: Granules have an affinity for acid dyes (acidophilic), therefore, stain well with eosin. Quite rare. Basophils: Basophils: Granules have an affinity for basic dyes (basophilic), therefore, stain well with hematoxylin. Very rare. Granulocytes are main portion of WBCs in humans, dogs, cats and horses

agile - affordable - accurate White Blood Cells (WBC) 2 - Agranulocytes Neutrophil Monocyte Lymphocytes are the main portion of WBCs in ruminants and pigs. B-lymphocytes, T- lymphocytes and Nature Killer Cells belong to this population. Monocytes are the biggest blood cells

agile - affordable - accurate Production of blood cells (hemopoiesis) Differentiation and maturation from proliferating stem cells occurs in the bone marrow Maturation of lymphocytes in takes place in the lymphatic tissue (lymphnods, spleen, thymus, etc.)

agile - affordable - accurate Haemostasis-Coagulation Coagulation occurs following tissue injury. Self enhancing and self limiting process in order to achieve localized, but effective reparation Several proteins (coagulation factors),Ca 2+ and thrombocytes are involved in the process As a result thrombocyte plugs and fibrin clots are formed It can be hindered by anticoagulants (EDTA, Heparin, Hirudin, etc.)

agile - affordable - accurate Anomalies in pathological conditions Elevated RBC number: compensatorical change in several diseases (heart failure, respiratoric insufficiency, etc.) Decreased RBC number: anemic state caused by bleeding, hemolysis, metabolic diseases, hemoglobinopathyes Decreased hematocrit: bleeding, hemolysis Elevated reticulocyte %: regenerative anemia Changes in Hemoglobin concentration, MCV and MCHC: anemia caused by iron or vitamin deficiency or hemoglobinopathies Elevated WBC numbers: inflammation, leukemia Decreased WBC numbers: immunosuppressive agents and diseases, bone marrow disfunction