Morphological Study of Human Blood for Different Diseases

Blood is survival sustaining fluid that flow through the whole
circulatory system .blood carries nourishment, electrolytes,
vitamins, hormones, oxygen, heat, antibodies...


Introduction
Normocytic cells that have normal size while Normochromic cell have the normal concentration of hemoglobin. Different type of abnormalities in the membrane of cell also change the shape of cell.
A generic term poikilocyte represent red blood cell with irregular shape, it can be classified by definite shape changes some are diagnostically significant and while some others are non-specific.
White blood cell also called Leucocytes which is the part of the immune system and perform function in immune responses.
They make up less than 1% of the cells in blood and involved in recognizing and neutralizing invaders such as virus and bacteria.
The leucocyte contains normal nucleus and mitochondria. Under the microscope they come in five major types and divided into two groups named for their appearance [6]. White blood cell are divided into two categories on the base of chemical characteristics and structure, the granulocytes that contain the granules which are bounded by cytoplasmic membrane and the agranulocytes, that lack obvious granules. Granulocytes include neutrophils, basophils, and eosinophils. Neutrophils form the 50-70% of white blood cell population and are most numerous of White blood cell.
Two types of chemicals are released by basophils, histamine and heparin. In allergic reactions histamine is involve and heparin is involved in anticoagulant. While to fight with parasitic worm eosinophil is responsible. Eosinophil release toxin that involved in inflammatory response and kill the worms. Agranulocytes include monocytes and lymphocytes, their cytoplasm is without granules [6]. The morphological features of white blood cells are less specie dependent then erythrocytes [4]. Platelets was discovered by Giulio Bizzozero in 1882 [7]. Thrombocytes are small blood cells 1.5-3μm in diameter and play an important role in hemostasis and in coagulation mechanism. Morphological abnormalities in platelets are found in hematological diseases such as myeloproliferative syndromes and acute mega karyoblast leukemia.
But for many decades the multifunctional and dynamic nature of platelets remain a field of interest for biologist. Hemostasis or blood coagulation is not the only function of platelets. Platelets are highly sensitive to different types of disease states assigned it to be one of the most affordable markers [8]. The mature platelets are 2-3μm in diameter and for 5-9 days remain alive. Approximately 1/3 of platelets is stored in spleen and 2/3 circulates in blood. The normal count of platelet is (150-400) multiply 10 to the power 3 per microliter of blood. Plasma membrane of thrombocytes contain phospholipid bilayer which is the site of expression of surface receptor and lipid rafts which are helpful in intracellular trafficking and signaling [9].
Platelets have two storage granules alpha and dense granules which perform the function of storage of biologically active molecules involved in the recrution of other cells during inflammation and initiation of coagulation [10]. Platelets play an important role in defending viral pathogens. Platelets secrete chemokines after activation, that attract the immune cells which help in the finishing of virus platelets has antimicrobial function, secret antimicrobial molecules including platelet microbicidal peptides and kinocidins.
These anti-microbial molecules kill pathogens. Another defending role of platelets is finishing of virus by phagocytosis [11]. Platelets are the smallest cells and in thrombus formation one of the main players. They undergo a series of biochemical and morphological changes. Platelets have the ability to bind to non-endothelial surface, bind to other platelets, secrete substance store in internal granules.
Platelet indicating differential dysfunction, aggregation, hyperactivation, and are capable of expressing the character of disease. Platelet biomarker is an important impact on the global scene of clinical application and development [12]. Blood is composed of about 55% of plasma. It is a light-yellow liquid and carries water, salt and enzymes. Plasma primary purpose is the transport of protein, hormones and nutrients to body parts.
Blood cells deposit their waste products to plasma and it helps in removing it from the body [13]. When all types of blood cell such as erythrocytes, leukocyte and platelets are separated from the blood, plasma is derived. They have 90-92 % water, plasma including constituents have electrolytes i.e. sodium, chloride, potassium, magnesium, bicarbonate and calcium. The blood wastes collected from blood cells are transported to kidney for excretion. Plasma containing 6-8% fibrinogen. Fibrinogen is circulating in the blood is transformed to fibrin when blood clotting is started which in turn helps to form constant blood clot [14].
Plasma origin is interesting because no organ produces it. It forms from water and salts absorbed through digestive tract [15].
A condition in which lack of mineral iron in the body commonly caused a disorder known as anemia. Body needs iron for protein hemoglobin. Iron helps erythrocytes in the transport of oxygen to different parts of the body. Thalassemia is an inherited disorder caused by genetic mutation that prevent the normal production of hemoglobin [16].

Study Area
The current study was carried out in district Swat with  Table 1).

Materials
The materials that are used in our study include a) For cleaning of fingertip and for the fixation of slides, For pricking of finger, disposable lancet is used. This questionnaire was given to the students before the collection of blood samples. During the collection of blood samples first we used ethanol to clean tip of ring finger and used the cotton to dry the finger from ethanol, then used the disposable lancet for pricking of finger. The lancet that once used for pricking were discarded to avoid its reuse.
Preparation of Blood Smears: The first drop of blood that comes after pricking the fingertip are discarded. The other drops of blood that comes from the prickled area were placed on the slide for the preparation of both thick and thin smear. Both the smears were prepared on one slide. Two clean slides were used, one for the preparation of smear and second were used for blood film.
Fixation of Blood Smear: The prepared smears were then dried in air for a small time. After this ethanol were used for the fixation of blood smears and allowed it in air for few minutes to dry.
When it became dry then dip it in water for removing of ethanol from slide.

Elliptocyte (ovalocytes)
Elliptocytes are elongated blood cells, vary in shape from egg shaped to rod or pencil shape. In normal blood very rare elliptocytes may be found. Elliptocytes increased in iron deficiency anemia. In current study elliptocytes were observed in 68 individuals ( Figure 4).

Bite cell
The bites are usually shaped like a semicircle but may also be irregular. Abnormal shaped erythrocyte with one or more semicircular portions detached from the cell periphery. The "bites" result from the deletion of hemoglobin with a distorted structure by specific cells "macrophages" in the spleen. Bite cell formation occur due to deficiency of GP6D (Glucose 6 Phosphate Dehydrogenase) and unstable hemoglobin. Hemolytic anemia associated with severe liver disease is another caused where bite cells are formed. In current study bite cells were found in 72 individuals ( Figure 6).

Keratocytes
They are fragmented erythrocytes. They come in a range of shapes instead of the pale-centered biconcave shape. They resemble a red cell with two horn like structure therefore also called 'horn' cell. It usually indicates an infection of the blood vessel walls, causes the membrane of some red blood cells to burst such as hemolytic syndromes. In current study keratocyte were observed in 8 individuals (Figure 8).

Sickle cell (Drepanocytes)
They are elongated red blood cells with pointed ends. It is formed by an inherited irregular form of hemoglobin. These hemoglobin tend to collect after unloading oxygen. In sickle cell anemia, the erythrocytes become rigid and have rod like structure. Due to irregular cell blood vessel may blocked causing tissue and organ damage. They are seen in sickle cell anemia, hemoglobin S/ beta-thalassemia. In current study sickle cells were observed in

Echinocytes (Burr cell)
Echinocytes are spiculated RBCs. They have multiple short,   and he moglobinopathies, such as hemoglobin C and unstable hemoglobin's. In current study irregular contracted cell were observed in 32 individuals (Figure 11).

Boat Shape Cell
The cell body may be dense or normal but retains central pallor.
These cells have sharpened ends. In some cells it may also be slit like. these cells are formed in the presence of abnormal hemoglobin and also associated with sickle diseased. In current study boat shape cell were observed in 13 individuals ( Figure 12).

Rouleaux
It is a bunch of red blood cells. Rouleaux formation may occur due to the presence of large amount of plasma protein in blood such as fibrinogen or immunoglobulins. condition that caused rouleaux formation include infections, inflammatory and connective tissue disorders and cancer. In current study rouleaux were observed in 16 individuals (Figure 13).

Acanthocyte (spur cell)
They are spiculated cells with irregular, pointed or club like projections that are unequally distributed on the cell surface.
Central polar is absent. It has spiked cell membrane, due to abnormal thorny projections. It forms as a result of membrane lipid abnormalities. During current study acanthocytes were observed in Figure 14: Acanthocyte (spur cell).

Discussion
During the current study the results represents 12 different type of human blood cells based on morphological size variation yielding tear drop, bite cell, boat shape cell, echinocytes, schistocytes, elliptocytes, sickle cell, irregular contracted cell, rouleaux cells, keratocytes, hypochromic cells and acanthocytes.
Mortimer [16] reported that in the marginal blood smear of two patients with Heinz bodies hemolytic anemia morphological abnormal red blood cells were observed, that abnormal cell were termed as bite cell because of their appearance of a piece bitten out. which is the risking factor for microcytic anemia. Shochienin [20] worked on echinocytes stomatocyte transformation and shape control of human erythrocyte.
They conclude that the erythrocyte has an energy needy shape, In the analyzed slides the ratio of bite and teardrop cells is higher. The characteristic of tear drop is myelofibrosis which is also seen in severe iron deficiency, megaloblastic anemia and thalassemia. Bite cell is produced due to the deficiency of glucose s6phosphate dehydrogenase and risking factor for hemolytic anemia.
Morphogenetic character is physical character of an individual.