Analysis Of Blood Smears

Analysis Of Blood SmearsThe aims of this experiment ar to study the morphology and characteristic of family mention the disproportion of inception when suffering from diverse illnesss and identified the differences between animal and humans phone line. To achieve the aims, smears of horse dividing line atomic number 18 inclined(p) and comp be with human blood. thusly the prepared human blood smears are observe under electron microscope.INTRODUCTIONBlood performs a lot of important authoritys within the personify it contributes homeostasis to the body and playing major role in defence system by phagocytises activity. On an average male adult who weights 70kg has a blood volume of astir(predicate) 5 litres, about 1/12th of the body weight. Blood consists 55% of blood germ plasm 45% of hematocrit in men, 58% blood plasma and 42% of hematocrit in women. Hematocrit packed with erythrocytes, leukocytes and platelets (Sherwood 2010).Erythrocytes are the most abundant blood cells with about 4-6 millions/mm3 in blood. Erythrocytes are comm just known as red blood cells. In mammalian, erythrocytes are free of nucleus to allow more room for haemoglobin and are biconcave in human body. Hence, vertebrates erythrocytes have a nucleus. Haemoglobin is the main contained in erythrocytes it carries oxygen to the tissues, collects and transports the unwanted carbon dioxide away, conveys nutritive substances like amino acids, sugars and mineral then gathers the waste materials that want to eliminated through the renal filter, carries hormones, enzymes and also vitamins to their sites of action (Sherwood 2010).Leukocytes or white blood cells are much less abundant than red blood cells but bigger in size. They obligated for the defence of organism or eliminate harmful foreign material and make up the immune system of the body. The density of leukocytes in the blood is 5000-7000/mm3. in that respect are both categories of leukocytes which are granulocytes and ag ranulocytes. Granulocytes is due to the presence of granules in cytoplasm and agranulocytes is the absent of granule in the cytoplasm. The granules are difference in different types of granulocytes and make it easier to do it among them. The granulocytes distinguish themselves as neutrophil, bromeosinophil and basophil. Agranulocytes distinguish themselves as lymphocytes and monocytes. Beside of the granules, shape of the nucleus help in recognition of leukocytes (Underwood 2004).The proportion of neutrophil amongst leukocytes is about 50-70%. Its main function is phagocytes bacteria and always present in large amount within the pus of wound. Unfortunately, these cells dead after phagocytes due to unable to renew the lysosomes that used in digesting microbes. Well, eosinophils only 2-4% amongst leukocytes, they attack parasites and phagocytes antigen-antibody complexes. Basophil is 0.5-1%, it secrete anti-coagulant and vasodilator substances as histamines and serotonin. It takes p art in phagocyte activity but the main function is secreting substances that mediate the hypersensitivity reaction. lymph cell own 20-40% proportion of leukocytes, its little cell that compact with round nucleus. Lymphocytes populate the lymphoid tissues (Bajanowski 1997), lymphoid organs (thymus, spleen, lymphoid nodules, and palatine tonsils) as well as the lymph that circulate in the lymphatic vessel (Underwood 2004).Monocytes cooperate in immune defence although they are only 3-8% of leukocytes volume and its the precursors of macrophages (Sherwood 2010). They are large blood cells, which originate in the bone marrows before enter to the blood circulation and they only stay for 24-36 hours then will immigrate into the connective tissue, where they hold up macrophages and move within the tissues. Monocytes migrate very rapidly to site if presence of an inflammation and intense phagocytory activity. Beside phagocytory activity, monocytes involve in secreting lysozime, interfere ons and new(prenominal) defensive substances (Underwood 2004).Platelets or thrombocytes are fragments of cells in the blood with diam about 2-3m hence they are much niceer than erythrocytes. Their density in the blood is only 200000-300000/mm3. They are responsible for blood clotting to delay blood loss from broken vessels. The blood vessel constricts to reduce blood flow and loss. Platelets then aggregate at the point of the broken vessel and create a plug to stop blood loss. To this purpose, they aggregate and release serotonin to reduce the diameter of lesion vessel and slow down the haematic flux to gain the blood coagulation (Sherwood 2010).Plasma is the most abundant liquid component of blood with a yellowish colour. It makes up approximately 55% of total blood volume. Plasma is alkaline and it functional to maintains the pH of the blood at approximately 7.4. It also maintains the osmotic balance of body cells. The composition of plasma is 90% irrigate and 10% of dry ma tter like glucose, lipids, protein, glycoprotein, hormones, amino acids and vitamins (Sherwood 2010).The morphology and characteristics of blood will be study by preparing the horse blood smears samplings that with and without dye. Blood smears stained by haematoxylin and eosin are easier to identify under microscope during this experiment. Blood smears of different pathologies will be investigated and identified by taking noted the numbers of cells present, shape and sizes of different types of cells and remark with drawing.METHODSFirst part of this experiment involved preparation of horse blood smear samples. A small drop of horse blood is placed at one end of a drop away and placed a cover slip at the edge of the blood then dragged light through the slide in range to produce a thin blood smears. The blood smear needs to be essentially thin until the blood is laboredly visible this is to ensure that individual cells were easily determined. If the smear front red that mean i t is not thin enough or too thick, this may be hard to observe through the microscope and do the cells count as packed cells is hard to see clearly under microscope.Second slides are prepared by using exactly the same way as the first one. Both slides placed immediately into a container containing ethanol for 2 minutes. Ethanol is a colourless substance and used as a fixative, it helps to preserve cell smear samples so that cells do not denature. It does not damage the cells at all, bonny helps to maintain them for analysis. After the use of ethanol, the slide then dried by just slanting it on a piece of tissue. Dap and rubs are not allowed, as it will destroy the thin film of smear. The unstained smear was considered ready for analysis. It was placed a side waiting for investigation progress.The second remaining slide then stained with haematoxylin and eosin. Haematoxylin is widely used in medical diagnosis it is a blue substance stain that used to stained nuclei of cells into bl ue or purple colour. The nuclear dapple is followed by other structures of the cells bodies with eosin stain that stain the granules of the cytoplasm in shades of red, pink and orange. Stained process performed by dipped the slide in a staining container containing haematoxylin for 2 minutes and rinse gently with water followed by dipped in another staining container containing eosin for 30 seconds and again rinse gently with water.A drop of mountant is applied on the smear and then covered with a glass coverslip. Mountant is a medium used for mounting a slid for microscopy purposes. The staining times varied slightly because the specimen was leave in the haematoxylin longer when the colour looks pale or pink and leave in the eosin for longer when it looks very dark blue.Both slides are completed and viewed under microscope. Unstained and stained smears were then observed under the microscope initiate by x10 magnification to find the cells and upgraded to higher definition of x40 m agnification for details bill. Observation started with stained smears followed by unstained smears, as stained smears is easier to determine the cells. Both smears were drawn accordingly and labelled all the particular structures of interest. Commend are made upon on how the stained smear differs from unstained smear.In second part of the experiment, human blood smears are observed. Stained human blood smears taken from patients who suffer from no known pathology, reaping hook cell anaemia, eosinophilia, corking lymphocytic leukaemia and iron deficiency anaemia were observed. A textbook includes of brief description and expectation of what to see from the pathological blood smears are provided during the practical.In this session, apiece slide provided is observed under microscope. Always started with x10 magnification and moved to x40 magnification while drawing. First, normal human blood smear is observed in order to identified elements in normal blood, then go onto the path ology smears and compared found morphology that identified in horse blood in part A. The cellular elements of all(prenominal) smear were drawn, labelled and recorded any differences observed in pathological smears when compared to normal blood smears. The relative numbers of each cell type are counted.RESULTSPart A Horse blood smearsFigure 1 illustrated stained horse blood smear under microscope of x40 magnification. The blood cells are stained with haematoxylin and eosin. unitary monocyte, one small lymphocyte, one neutrophil and bundle of erythrocytes (red blood cells) are seen. Nuclei of the leukocytes were purple-blue in colour due to the haematoxylin staining and the cytoplasm of the leukocytes appeared pink due to eosin staining. The erythrocytes are more abundant compared with leukocytes. Renown, erythrocytes are boconcave disc that absent of nuclei and mitochondria.Figure 2 shows the unstained horse blood smear. The blood cells appeared to be transparent and hard to determ ine the differences between the erythrocytes and most of the leukocytes except monocyte, as it is great in size.Part B Human blood smearsFigure 3 shows the human blood smear with no known pathology. Erythrocytes, leukocytes (neutrophils, monocytes, lymphocytes, basophil) and platelets are presented. The smear make out with more abundant of erythrocytes (R.B.C) than leukocytes and they all appeared healthy. The leukocytes were blue-purple in colour surrounded by numerous erythrocytes that were pinkish in colour. Most of the leukocytes seen are neutrophils this proves the theory stated leukocytes making up with 50-70% of neutrophils. The neutrophils were intermediate in size, lymphocyte was smaller and monocyte was larger. Their cytoplasm appeared pink in colour. The nucleus of neutrophil lob with clumps of chromatin.Figure 4 shows human blood smear with sickle cells anaemia. Abnormal red blood cell morphology and sickle cells are seen.Figure 5 shows human blood smear with eosinophi lia deficiency. Abnormal or sickle red blood cells appeared. eosinophile and monocytes are broken. Several of dim cells presented.Figure 6 shows the blood smear for acute lymphocytic leukaemia. The erythrocytes are not as densely pack as in the smear of human blood with no pathology. This observation clearly illustrated the presented of several lymphocytes in the smear and it appeared larger than erythrocytes. Where acute lymphocytic leukaemia is a blood cancer where the body produces a large numbers of lymphocytes.Figure 7 shows the smear for human iron deficiency anaemia. The erythrocytes are pallor in appearance. Some of the erythrocytes were larger in size. anyway that, smudge cells and different types of leukocytes seen in this smear. There are lymphocytes that small in size and also neutrophils.DISCUSSIONMany diseases, disorders, and deficiencies can distinguish by observation of blood cells distribution and appearances (Bain 2005). Disproportionate numbers of leukocytes, pr esence of immature leukocytes, too high or too low of platelets counts, and deformed of red blood cells are all signs of serious diseases. Somehow, blood smear provides the primary evidence of a specific diagnosis. Monocytes of horse blood smear in fig. 1 are greater in size compare with human blood smear in fig.3. The comprehensive kinetic force between erythrocytes of horse blood is stronger and produced closely link long chain of erythrocytes. The erythrocytes in fig.1 and 3 appeared normal, uniformed in size and do not have a nucleus as most other cells do. They are round and flattened like a donut with a depression in the middle. Due high density of haemoglobin presented inside the erythrocytes (Sherwood 2010), they appear pink to red in colour with a pale centre. While there are some erythrocytes in fig.4, 5, 6 and 7 had significant different in shape and irregularities that indicate severe problems.The histological section with stained are more visible and can be noted that the nuclei of the cells appeared purple-blue with stained of haematoxylin (Bain 2005) cytoplasm appeared pinkish with stained of eosin. Unstained leukocytes are colourless and hard to determine as they lack haemoglobin (Bain 2005). The stains enhanced the illustration of the leukocytes and make it easier to distinguish. Granulocytes and agranulocytes were differentiated by observed their cytoplasm. Granulocytes are neutrophil, eosinophil and basophil that has granule in their cytoplasm and its cytoplasm is visible when staining, while agranulocytes are lymphocytes and monocytes that absents of granule in their cytoplasm where their cytoplasm appeared transparent although stained.Neutrophils are cells that have cytoplasm with pink granules, intermediate in size with lobed clumped nucleus, can be identified by observing their nuclei their nuclei are segmented into 2-5 lobed of different shapes. They composed majority of leukocytes and function to phagocytosis . Eosinophils will easily recognize with their large, red-orange granules. Unfortunately, they arent found from the smears because they are generally low in number. Eosinophils most often become elevated in number when the individual are facing with allergies or parasitic infections. Basophils (figure 1) had large black granules and least often seen from the smears as they are only 1% of leukocytes. Increased numbers of basophiles are not often encountered but may be elevated in certain leukaemia, chicken pox, ulcerative colitis, or after an immunization. Monocytes are the largest cell amongst leukocytes with diameter of 12-20 m and are often referred as phagocytes. They engulf particles such as cellular debris and bacteria. LHYPERLINK javascript optionsdisplay(../../../glossary/lymphocyte.html)ymphocytes are smaller and have a homogeneous cytoplasm and a smooth, round nucleus. These cells are responsible for the production of antibodies or immunoglobulin (Bajanowski 1997). There are two types of lymphocyte s, B and T cells and they mediated within each other. B cells induce production of antibodies T cells destroy specific cells (Bajanowski 1997).Figure 4 illustrated human sickle cell anaemia. Sickle cell anaemia (SCA) affects millions of people worldwide (Charlotte 2010). SCA is disorders of erythrocytes that caused difficulty to haemoglobin molecules when delivers oxygen to cells throughout the body (Peterson 2009). The change of the amino acid results in haemoglobin that responds to the oxygen deficiency by stacking filaments and clustering in red blood cells containing the mutated protein in such a way that their shape is distorted (Sherwood 2010).Eosinophil usually hardly noticeable in blood smears indicates the response of the body to abnormal cells, parasites, or substances that cause an allergic reaction. Donor of the blood smear illustrated in figure 5 may have eosinophilia disorder as broken eosinophil is presented. Eosinophilia is commonly happened to people who have asthma , hay fever, food allergic or parasitic infections such as intestinal worms (Sherwood 2010).In the acute lymphocytic leukemia sample shown in figure 6, there was a noticeable increase in the number of lymphocytes seen. The erythrocytes are pallor and lymphocytes appeared larger than erythrocytes and this is due to a disease of lymphoid cells causing uncontrolled production of lymphocytes (Underwood 2004). Acute lymphocytic leukaemia is a disease where the physical changes take place within the cell (McClain 1990), a reduced count of red blood cells with a raised level of leukocytes. This may leads to an collecting of blast cells in the bone marrow and causes bone marrow failure (McClain 1990).All the red blood cells in the iron deficiency anaemia sample appeared pale in colour. This usually caused to people with poor diet that contains little iron especially vegetariansbecause the main dietary source of ion is red meat. Besides that, diseases of the small intestine such as gluten i ntolerance can reduce its ability to absorb iron (Sherwood 2010).