Haematology

1. Haematology Unit 21 P4, M3, D3

2. A B C D E

4. Blood Components • Plasma • Red Blood cells • Leucocytes • Platelets • Clotting Factors What are the functions for the different components?

5. plasma (55%) red blood cells (5-6-million /ml) white blood cells (5000/ml) platelets

6. x 1000

7. Plasma liquid part of blood plasma transports:- • soluble food molecules • waste products • hormones • antibodies

8. Erythrocytes (Red blood cells (RBCs)) • transport oxygen • specialised to do this Also carry some CO2

9. Leucocytes (White blood cells) • the bodies “defence” • part of the immune system • much larger than RBCs • far fewer • have a nucleus • 4000-13000 per mm3

10. Leucocytes (White blood cells) • 2 types • granulocytes and the agranuloctyes • The granulocytes have granules in the cytoplasm and absorb different types of stains. (dyes) • ALSO known as phagocytes and lymphocytes

11. Platelets if you get cut:- • platelets produce tiny fibrin threads • these form a web-like mesh that traps blood cells. • these harden forming a clot, or "scab." • 150,000 to 400,000 per mm3

12. Erythrocytes specialisations 1) biconcave shape 2) no nucleus  extra space inside 3) contain haemoglobin  the oxygen carrying molecule  250million molecules / cell increases the surface area so more oxygen can be carried

13. Haemoglobin • gives red blood cells their colour • can carry up to 4 molecules of O2 • associates and dissociates with O2 • contains iron

14. Function of Haemoglobin When there is a high concentration of oxygen e.g in the alveoli haemoglobin combines with oxygen to form oxyhaemoglobin. When the blood reaches the tissue which have a low concentration of oxygen the haemoglobin dissociates with the oxygen and the oxygen is released into body tissues

15. Agranulocytes • Leukocytes without granules are agranulocytes • The two types are monocytes and lymphocytes • lymphocytes are smaller of the two.

16. Phagocytes • Monocytes and macrophages • Provide a non-specific response to infection • http://www.microbelibrary.org/images/tterry/anim/phago053.html

17. Monocytes

18. Lymphocyte

19. Lymphocytes Provide a specific immune response to infectious diseases. There are 2 types: - - T-cells - B-cells They produce antibodies.

20. Granulocytes • Most have irregular, lobed nuclei. • The cytoplasmicgranules contain enzymes involved in detoxification of foreign substances, blood clotting, and various immune responses • Neutrophils with one or two lobes are immature or abnormal

21. Blood Can you? • State the composition of Blood • State the function of red blood cells and plasma • Explain the function of haemoglobin in the transport of oxygen • State the function of macrophages and lymphocytes

22. Taking a bloodsample • Use role play to demonstrate how to take a blood sample. • Answer these question... • Explain the use of • Tourniquet • Alcohol • Antiseptic • Applying pressure to the area after removing the needle. • Create a cartoon strip that illustrates an easy to follow method.

23. Creating a blood smear. • Whilst watching the video, answer these questions. • Explain why the blood is spread using a capillary action of the spreader slide. • Identify the correct angle of the spreader slide. • How much blood is added to each slide? • Describe why the amount of blood added to each slide is important. • Explain why the slides were labelled using a pencil. • Explain why the slides are air dryed. • Explain why the slides are fixed with methanol.

24. P4Diagnostic techniques When research P4 include the term ‘clinical method’ in your search. Journals will be the most appropriate sources online. Websites such as webmd do not go into detail of the laboratory test. • blood smears • Haemocrit • Blood cell count • reticulocyte count • Erythrocyte count • platelet counts • mean corpuscular Volume • analysis for iron deficiency • tests for abnormal haemoglobin • Blood volumes • coagulation

25. Identifying Blood Type • Red blood cells have antigens of their surface. • A, B or neither • Rhesus D • Blood has antibodies to detect foreign red blood cells detecting antigens on RBCs.

26. When antibodies detect an antigen they cause agglutination- RBCs clump together

27. Blood Cell Count (c) describe the use of a haemocytometer to count the numbers of erythrocytes and leucocytes (to include details of dilution);

29. Hemocytometer • Originally used in counting RBCs • Consists of a thick glass microscope slide with a rectangular indentation that creates a counting chamber • Counting chamber - engraved with a laser-etched grid of perpendicular lines • Raised edges hold cover slip above these marked grids

31. Hemocytometer • Counting chamber is a 3 x 3 mm square and divided into nine large squares • Each resulting square measures 1 mm2, which are used for white blood cell counting (Square A) 1 mm2

32. Hemocytometer • Counting chamber is a 3 x 3 mm square and divided into nine large squares • The center square is divided into 25 smaller squares. • (1/25 mm2) 1/25 mm2

33. Hemocytometer • Each smaller square is subdivided into 16 even smaller squares, which are used for counting red blood cells. • (Square B = 1/400 mm2) 1/400 mm2

34. When you count ……NORTH-WEST Rule • Establish a rule to avoid counting cells twice or not at all Example: • Count only cells that lie on the NORTH (top) and WEST (left) hand lines of each box • the ones on South East lines will be counted with next box when you get to them

36. ` • Count in triple lined (central grid) • 1st count cells in at least 5 sqs and calculate average • Calculate concentration of cells in 1 ml. • Then try counting cells in larger number of sqs and repeat calculations • Extension • Do the same as above with 1 in 10 dilution

38. Research • Normal and abnormal concentrations of the different cell types in the blood. • Erythrocytes • Leucocytes • Different types of leucocyte • Reticulocyte • Platelets • Describe the conditions the abnormal cell counts cause.

39. http://www.proprofs.com/quiz-school/story.php?title=using-haemocytometerhttp://www.proprofs.com/quiz-school/story.php?title=using-haemocytometer

40. Each triple lined square has a volume of 0.004mm3 • We are counting five triple-lined squares so 0.004mm3 x 5 = 0.02mm3 • Our blood is yeast cells. This is diluted 100 times. • If the number of cells counted is E: • In 1mm3 of the original yeast sample the number of cells will be: • 1 / 0.02 x E X 100

41. Have a go!!

42. answer • 1) A quick count of the cells reveals that there are 39 present in the haemocytometer.2) Since the haemocytometer is comprised of 9 large squares, the average number of cells per large square is 39 9 =4.33 3) Each small square measures 0.25mm x 0.25mm, and so each large square must measure 1mm x 1mm. This is equivalent to 0.1cm x 0.1cm. The depth of a small square is the same as a large square and is given by 0.25mm, which is the same as 0.025cm. • Therefore a large square measures 0.1 x 0.1 x 0.025 cm, giving a volume of 2.5×10 −4 cm 3 , which is the 2.5×10 −4 ml.4) We have 4.33 cells per large square. Each large square has a volume of 2.5×10 −4 ml. Therefore the number of cells per ml is 4.33 2.5×10 −4 =17333 .5) 17333 cells per ml. Therefore in our sample of 100ml, we have 100×17333=1733300 = 1.73×10 6 cells.

43. Counting Erythrocytes and Leucocytes

44. Reticulocyte Count • A reticulocyte count measures the percentage of reticulocytes (slightly immature red blood cells) in the blood. • The test is done to determine if red blood cells are being created in the bone marrow at an appropriate rate. • The number of reticulocytes in the blood is a sign of how quickly they are being produced and released by the bone marrow.

45. Mean Corpuscular Volume RBC indices • Red blood cell (RBC) indices are part of the complete blood count (CBC) test. • They are used to help diagnose the cause of anemia, a condition in which there are too few red blood cells. The indices include: • Average red blood cell volume (MCV) :(haemocrit, haemocytometer) • Hemoglobin amount per red blood cell (MCH): (spectroscopy) • The amount of hemoglobin relative to the size of the cell (hemoglobin concentration) per red blood cell (MCHC)

46. Haemocrit • also known as packed cell volume (PCV) or erythrocyte volume fraction (EVF), is the volume percentage (%) of red blood cells in blood. • It is normally about 45% for men and 40% for women

48. Spectroscopy • Ultraviolet-Visible Spectroscopy: Measuring level of absorption or transmission of light. • When a calibration curve is completed the concentration of the sample can be calculated.

49. Erythrocyte Sedimentation Rate • Testing physical aspects of whole blood. • Anti-coagulated blood is put in a thin column and vertically aligned. • The blood is allowed to settle under influence of gravity. • The blood separates into plasma and the cells at the bottom of the tube. • The fall of blood cells is recorded at mm/h.

50. Erythrocyte Sedimentation Rate