Group 3: David Earl Dadivas Rose Ann Esteban Karyl Sabrina Javellana Methence Tayuba
Submitted on: September 13, 2006
LABORATORY ACTIVITY BLOOD AND HEMOPOIESIS PROCEDURE: 1. Put a drop of blood on a clean slide. Using a second slide in contact with the first at 45°, smear the drop away from you until it evenly distributes between the 2 slides and leaves a thin film of blood on the first. If too much blood is used, the layer of cells left will be too thick, the leukocytes will be more or less obscured and will not stain properly. 2. Put a drop of Wright’s stain and allow to dry immediately. If drying is delayed, coagulation commences and the leukocytes will not be evenly distributed; RBC will develop small spinous processes or the “crenated” look.
3. Locate the cellular components of blood in your prepared specimen, or a prepared slide of human blood smear. 4. Under the oil immersion objective, focus on a section of a red bone marrow smear and look for the different cells described above. Note the appearance of the numerous cellular elements in their different stages of development. Note some fat cells seen among the blood cells. 5. Draw and label the histologic appearance of the following: a. RBC
b. Granulocytes: neutrophils, eosinophils, basophils
c. Agranulocytes: monocytes, lymphocytes
d. Red Bone Marrow and its cellular elements
6. Draw and label the stages of sequential hemopoietic development of each of the formed elements in the bone marrow.
STUDY – FOCUSING QUESTIONS
1. Compare and/or contrast the following by tabulating the required information: a. Types and Subtypes of Leukocytes Percentage in Total WBC Count
Shape and Visibility of Nuclei
GRANULOCYTES Neutrophils 55-65%
Eosinophils 1-3%
Basophils 0.5-1%
Lymphocytes 20-35%
Monocytes 3-8%
10-12 mm in diameter; consists of 2-5 lobes connected by thin strands of chromatin.
Has 2 oval lobes connected by a nuclear thread; no nucleolus.
Big, intensestaining round or slightly indented nucleus.
Oval or slightly indented nucleus is eccentrically located.
Round, spherical in vivo.
Round, spherical in vivo.
A thin rim of clear blue cytoplasm surrounds nucleus.
Abundant cytoplasm
Phagocytize bacteria and particulate matter in CT, then die and form pus.
Participate in allergic reactions by phagocytizing Ag-Ab complexes and by this dampen the allergic response Bone marrow
Heterochromatic nucleus is divided into irregular lobes, often obscured by overlying basophilic specific granules. Nucleus is U or S shaped. Cytoplasm is round, spherical in vitro; Filled with coarse, basophilic, membrane bound, water soluble granules. Involved in allergic and inflammatory reactions.
Immunological reponses of the body
After differentiating into a macrophage in the CT, it becomes phagocytic;
Bone marrow
Bone marrow and Plasma cells (BLymphocytes). Predominant
Stem Cells of Bone marrow
1. Long-lived lymphocytes survive for yearspredominate
100 days or more
Amount of Cytoplasm
Primary Function
AGRANULOCYTES
Bone marrow Origin Predominant Type in Circulation
Most predominant 8 days
Predominant 3-6 days
Least predominant 10-12 days
Life Span
Predominant
2. Short-lived lymphocyteshours/days/weeks
b. Types and Subtypes of Leukocytes Content and Staining Property of Specific Granules
GRANULOCYTES
AGRANULOCYTES
Neutrophils
Eosinophils
Basophils
Specific granules have low staining affinity and stain light pink ('neutral stain'). They contain phagocytins (bactericidal),
Specific granules stain pink with Wright’s stain and contains proteins not found in
Heparin and vasoactive compounds
Lymphocytes
Monocytes
collagenase, to help the cell move through connective tissue, and lactoferrin, which is toxic to bacteria and fungi.
Content and Staining Property of Azurophil Granules
Azurophilic granules contain lysosomal enzymes, defensins, and some lysozyme. They stain aviolet colour when prepared with Wright's stain. The enzymes produce hydrogen peroxide which is a powerful antibacterial agent.
other cell types: major basic protein, eosinophilic cationic protein (both aid in killing parasitic worms), and eosinophilicderived neurotoxin. Azurophilic granules contain lysosomes.
Azurophil granules can only be seen in small lymphocytes.
Fine Azurophil granules are scattered amongst a grayishblue cytoplasm.
Lipid mediators, like leukotrienes, and several cytokines, one of them being interleukin4.
Other Cellular Products
Immunologic Competencies Arising From Cytoplasmic Contents
c. A Differential Count of Blood Components
White blood cell count (WBC)
Red cell count (RBC)
Normal Values 4,300 and 10,800 cells per cubic millimeter (cmm)/ 4.3 10.8 x 109 cells per liter 4.2 - 5.9 million cells(cmm)/ 4.2 5.9 x 1012 cells
Clinical Significance of Deviation from Normal Values An increase or decrease in the numbers of the different types of white blood cells can help identify infection, an allergic or toxic reaction to certain medications or chemicals, and many conditions, such as leukemia. If the RBC count is low (anemia), the body may not be getting the oxygen it needs. If the count is too high (a
per liter
13 - 18 grams per deciliter for men and 12 - 16 for women (international units 8.1 - 11.2 millimoles/liter for men, 7.4 9.9 for women) 45 - 52% for men and 37 48% for women
Hemoglobin (Hb)
Hematocrit (Hct)
86 - 98 femtoliters
Mean cell volume (MCV)
27 - 32 picograms Mean cell hemoglobin (MCH) 32 - 36% Mean cell hemoglobin concentration (MCHC) 11 - 15 Red cell distribution width (RDW)
Platelet count
150,000 400,000/ cmm (150 - 400 x 109/liter)
condition called polycythemia vera), there is a risk that the red blood cells will clump together and block tiny blood vessels (capillaries). The hemoglobin test measures the amount of hemoglobin in blood and is a good indication of the blood's ability to carry oxygen throughout the body.
This test measures the amount of space (volume) red blood cells occupy in the blood. The value is given as a percentage of red blood cells in a volume of blood. The MCV shows the size of the red blood cells. This helps in the diagnosis of different types of anemia. MCH value is the amount of hemoglobin in an average red blood cell. This helps in the diagnosis of different types of anemia. The MCHC measures the concentration of hemoglobin in an average red blood cell. This helps in the diagnosis of different types of anemia. Red cell distribution width (RDW) indicates different sizes of red blood cells are present. If there are too few platelets, uncontrolled bleeding may be a problem. If there are too many platelets, there is a risk of a blood clot forming in a blood vessel.
d. Platelet Granules Location in Platelet Environment Contents
Function
Alpha Granules Chromomere of platelet Fibrinogen Plate-derived growth factor Other proteins
Delta Granules Chromomere of platelet Serotonin
Lambda Granules Chromomere of platelet Lysosomal Enzymes
Aid in platelet aggregation
Aid in platelet aggregation.
Contribute to clot removal
Fibrinogen may act as glue binding platelets together.
e. Components
Cascade Events of Formation
Aggregated platelets Platelet Plug Platelets Damaged blood vessels Plasma proteins Fibrin Thrombus
Small defects in the endothelial lining of blood vessels leads to aggregation on the damaged tissue. Factors from blood plasma, damaged blood vessels, and platelets promote cascade interactions of around 13 plasma proteins. The cascade results in the formation of the thrombus through the conversion of fibrinogen to fibrin by the enzyme thrombin.