Blood and blood cells

1

BLOOD AND BLOOD CELLS

PRESENTED BYDR HIMANSHU GUPTA

PG Ist YEAR

2

Introduction Plasma & SerumFunctions of bloodPlasma proteinsRBC’sErythropoiesisHemoglobinESR & PCV……WBC’s

CONTENTS

3

It is basically connective tissue in fluid form. It is called as the fluid of life, as it carries oxygen from lungs to all parts of the body and carbondioxide from all parts of the body to the lungs.

BLOOD

4

Color:  Scarlet red and Purple red

Volume:  In new born-450 ml                      Normal healthy male adult-5L                      Female-4.5L

3) pH: Slightly alkaline 7.44) Specific Gravity:    Total blood- 1.052-1.061                                    Blood cells-1.092-1.101                                    Plasma-1.022-1.026

5) Viscosity:  5 times more than water

5

COMPOSITION OF BLOOD

BLOOD

LIQIID PORTION

PLASMABLOOD CELLS

FORMED ELEMENTS

6

BLOOD CELLSRED BLLOD CELLS

WHITE BLOOD CELLS

THROMBOCYTES

7

Plasma is the straw colored liquid component of blood.

It contains 90-92% 0f water and 8-9% of solids.

These solids are the organic and inorganic substances.

The removal of coagulation factors from plasma leaves a fluid similar to interstitial fluid, known as serum.

PLASMA

8

SERUM

Within 45 minutes of clot formation serum oozes out of the clot

It is different from plasma only by the action of fibrinogen

Serum = Plasma - Fibrinogen

9

10

FUNCTIONS OF BLOODNutrient supply

Respiratory Function

Excretory Function

Transport Of Hormones And Enzymes

Regulation of Acid-Base Balance

Regulation of body Temperature

Storage Function

Defensive function

11

In embryonic stage plasma cells are synthesized by mesenchymal cells where as in adults they arise from reticuloendothelial cells of liver and also from spleen, bone marrow.

Plasma proteins

12

•Role in blood coagulation( Fibrinogen).

•Role in Defense Mechanism: γ-globulins act as antibodies, also called immunoglobulins

•Role in transport mechanism: Albumin,α and β-globulins are responsible for the transport of hormones and enzymes

Functions Of Plasma Proteins

13

•Role in viscosity of blood: Plasma proteins provide viscosity which is essential for maintaining BP. Albumin provides maximum viscosity.

•Role in ESR: globulin and fibrinogen accelerate rouleaux formation, which in essential for ESR.

•Role as reserve proteins: Act as last source of energy in case of starving or inadequate food intake

14

RED BLOOD CELLSRBCs also called as erythrocytes are non nucleated formed elements of the blood.

There red color is due to hemoglobin

Normal Values: Males: 5 million/cu mm Females: 4.5 million/cu mm

NORMAL VALUES

15

SHAPE AND SIZE• Normally disk shaped & bi-

concave.• Central portion thinner and

periphery thicker.

• Size: Diameter = 7.2μ (6.9-7.5 μ)

Thickness=• At periphery=2.2 μ • At center = 1 μ

16

Rouleaux formation

When blood is taken out, RBC pile up one above another like pile of coin. This property of the red blood cells is called rouleaux formation.

17

Transport of oxygen from lungs to tissues: Hemoglobin + oxygen = oxyhemoglobin

Transport of carbon dioxide from tissues to lungs:Hemoglobin + carbon dioxide = carbhemoglobin

In determination of blood groups:Carries blood group antigens like antigen A,B etc. that help in determination of blood groups & enables to prevent reactions due to incompatable blood transfusion

Functions of RBC’s

18

Variations in Number of cells

1INCREASE IN NUMBER

2DECREASE IN NUMBER

19

Increase In Cell CountPHYSIOLOGICAL VARIATIONS An increase in the red blood cell count is known as

polycythemia.

1. AgeAt birth, the red blood cell count is 8-10 millions/cu mm of blood. The count decreases within 10 days after birth due to destruction of cells causing physiological jaundice in some infants.

20

2. SexBefore puberty and after menopause in females the RBC countis similar to that of males. 3. High altitudeThe inhabitants of mountains have an increased red blood cell count of more than 7 millions/cu mm.

21

4. Muscular ExerciseThere is a temporary increase in red blood cell count after exercise.

5. Emotional ConditionsThe red blood cell count is increased during the emotional conditions like anxiety.

22

6. Increased Environmental TemperatureThe rise in the atmospheric temperature causes elevation of red blood cell count.

 7. After MealsThere is a slight increase in the red blood cell count after taking meals.

23

Decrease in cell count

1. High barometric pressuresAt high barometric pressures, when the oxygen tension of blood is higher than the normal, the red blood cell count is decreased. 2. During sleepThe count of red blood cells is slightly reduced during sleep.  3. PregnancyIn pregnancy, the red blood cell count is less because of increase in extracellular fluid volume.

24

Pathologic Variations

PATHOLOGICAL POLYCYTHEMIA

RBC count >7 millions/cu mm of the blood. Types:

1) Primary Polycythemia 2) Secondary Polycythemia

25

PRIMARY POLYCYTHEMIA - POLYCYTHEMIA VERA

Polycythemia Vera is a disease with persistent increase in red blood cell count above 14 millions/cu mm of blood.

This is always associated with increased white blood cell count above 24,000/cu mm of blood.

Polycythemia vera occurs in disorders like malignancy of red bone marrow.

26

SECONDARY POLYCYTHEMIA

This is secondary to some of the pathological conditions (diseases) such as:

• Respiratory disorders like emphysema. • Congenital heart disease.• Chronic carbon monoxide poisoning.• Poisoning by chemicals like phosphorus and

arsenic.

27

Variations In Shape Of RBC1. CRENATION-shrinkage as in hypertonic solution2. SPHEROCYTOSIS-Globular form as in hypotonic

solution.3. ELLIPTOCYTOSIS-Elliptical shape in certain anemia's4. SICKLE CELL-cresent shape as in sickle cell anemia5. POIKILOCYTOSIS-unequal shape due to deformed cell

membrane. the shape can be flask, hammer or any other unusual shape.

28

Variations In Structure Of RBC

1. PUNCTATE BASOPHILISM-dots of basophilic material(porphyrin) appear in the red blood cells giving a stripped appearance.

This occur in conditions like lead poisoning2.RING-a ring or twisted strands of basophilic material

appear in the periphery of the RBCs.It is also called as Cabots ring.

It is present in certain type of anemia.

29

3 HOWEL –JOLLY Bodies-in certain anemia's some nuclear chromatin fragments are present in RBCs.(in splenectomy pts.)

4- HEINZ-bodies- they are seen in G6PD deficiency. They represent denatured globin chains when there is not enough G6PD around, the bonds between heme and globin are attacked. Heme is just recycled, but globin chain become denatured, forming a little ball that stick to the inside of red cell membrane.

30

31

Life span of R.B.Cs•Average life span of RBCs is about 120 days. They are destroyed in reticuloendothelial system.•When cell became older the cell membrane became fragile.•Destruction occurs mostly in capillaries of spleen.•Spleen is called as GRAVEYARD OF R.B.C.s

32

Hemolysis & Fragility of RBC’sHemolysis: destruction of formed elements of blood/breakdown of RBC’s to liberate hemoglobin

Fragility: it is the susceptibility of RBC’s to hemolysis

33

Process of hemolysis

Normally plasma and RBC’s are in equilibrium, but when this equilibrium is disturbed the cells get effected.

Conditions when haemolysis occurs:•Hemolytic jaundice•Antigen-antibody reactions•Poisoning by chemicals/toxins

34

Haemolysins

These are the substances that cause destruction of RBC’s

These can be of 2 typesChemical substances : alcohol ,benzene, chloroform , chemical poisons(nitrobenzene)Substances of bacterial origin: toxins from bacteria and venom of poisonous snakes

35

Fate of RBC’s

Destruction of RBC’s in spleen

Release of hemoglobin

Globin

Protein pool

Stored & reused

Iron + Apoferritin

Ferritin

Stored & reused

Porphyrin

Bilirubin

Excreted

36

Erythropoiesis

It is the process of origin, maturation and developmentof erythrocytes.

Hemopoiesis is the process of origin development and maturation of all the blood cells.

37

Sites of ErythropoiesisIN FETAL LIFE: erythropoiesis occurs in different sites atDifferent periods-• Mesoblastic stage: during first trimester of

intrauterine life, RBC’s are produced from mesenchymal cells of the yolk sac.

• Hepatic stage: during second trimester, RBC’s are produced from the liver. Some cells are also produced from spleen and lymphoid organs also.

• Myeloid stage: during third trimester, RBC’s are produced from red bone marrow & liver.

38

IN NEW BORN BABIES AND ADULTS:

• Upto the age of 20 years: bone marrow of all bones

• After the age of 20 years: all membranous bones and ends of long bones

39

40

Stages of Erythropoiesis

41

42

Hemoglobin•Hemoglobin - Structure Content : It is composed of the protein globin (a polypeptide), and the pigment heme.• The hemoglobin has the ability to combine with oxygen is due to the four iron atoms associated with each heme group within the molecule.• A heme group consists of an iron (Fe) ion held in a heterocyclic ring, known as a porphyrin. •This porphyrin ring consists of four pyrrole molecules cyclically linked together (by methine bridges) with the Iron ion bound in the centre.

43

•Hemoglobin (Hb) is a chromoprotein.•Molecular weight 64,458 Dalton•Each hemoglobin molecule caries four molecule of oxygen and each gram of hemoglobin can carry 1.34 ml oxygen.•About 6.25 grams of hemoglobin is synthesized each day to replace the hemoglobin lost due to normal destruction of RBCs•Synthesis begin from proerythroblast to reticulocyte.

44

45

Hemoglobin content:Average Hb content of blood is= 14-16gm/dl.

Varies with age and gender.

At the time of birth, in infants and growing children the RBC count is more and so is the hemoglobin content .

In adult males-14-16gm%

In adult females-12-14.5gm%

46

Hemoglobin types:

Two types : Adult (Hb A): the globin contains two alpha and two beta chains and has less affinity to oxygen

Fetal (Hb F):there are two alpha and two gama chains and has more affinity for oxygen

47

METHODS OF ESTIMATION OF HEMOGLOBINColour basedBased on color of hemoglobin or derivative of hemoglobinPhysical methodBased on specific gravityChemical methodBased on iron content of hemoglobinGasometric methodBased on oxygen combining capacity of hemoglobin.Spectrophotometric methodBased on measurement using spectrophotometric devices.

48

49

SAHLI MEHOD PRINCIPLE

Hemoglobin +0.1 HCL Acid hematin(brown color)

GLOBIN

The resulting color is diluted with water and matched with brown color glass.

50

51

52

53

54

55

56

E S R

ESR is the rate at which the erythrocytes settle down. When mixed with an anticoagulant and allowed to stand undisturbed in a vertical tube, the RBC’s settle down due to gravity With a supernatant layer of clear plasma.

57

Two ways of determinationWestergren’s Method:

i. Westgren’s tube is used.ii. 300 mm long tube, opened on both ends.iii. Marked from 0-200 mm from above downwardsiv. 1.6 ml of blood is mixed with 0.4 ml of 3.8%

sodium citerate.(4:1)v. The blood is loaded up to 0 mark above the tubevi. The reading is taken thereafter.

58

Wintrobe’s Method:

i. Wintrobe’s tube is usedii. It is a short tube, opened at one endiii. It is 110mm long with 3mm boreiv. Determines ESR & PCVv. It shows marking on both sides, from 0-100(ESR)

&100-0(PCV)vi. 1ml of blood is mixed with ethylene-di-amine-

tetra-acetic acid(EDTA)vii. Blood is loaded up to 0 mark above

59

Normal values of ESRBY WESTERGREN’S METHOD:In Males = 3-7 mm in 1 hr.In Females = 5-9 mm in 1 hr.Infants = 0-2 mm in 1 hr.

BY WINTROBE’S METHOD: In Males = 0-9mm in 1 hr.In Females = 0-15mm in 1 hr.Infants = 0-5 mm in 1 hr.

60

Pathological VariationsRaised in following conditions:Tuberculosis

•Anaemia

•Malignant Tumors

•Rheumatoid Arthritis

•Rheumatic Fever

•Liver Diseases

Decreases in following conditions:

•Allergic Conditions

•Sickle cell anaemia

•Polycythemia•Leucocytosis•Hereditary spherocytosis

61

62

P C V

Volume of RBC’s in blood that is expressed in percentage.

Also called as Hemocrit value

63

Method of Determination

64

In Males = 40-45%In Females = 38-42%

Normal Values

65

White Blood Cells

• Also known as WBC’s or leucocytes

• Colorless and nucleated formed elements of blood

• Comparatively large in size and lesser in number

• Play a role in defense mechanism of body

66

67

68

69

NEUTROPHILS•Also known as polymorphonuclear leucocytes becausethe nucleus is multilobed

•The number of lobes varies from 1-6

•With Leishman’s stain, the granules stain equally

Electr. Micros. view

70

•Granules appear violet in color

•Diameter= 10-12μ

•Ameboid and phagocytic in nature

Along with monocytes neutrophils provide Ist line of defense.

They wander freely through out the body.

71

Mechanism of action:

Released in large number from the blood.

Move by diapedesis to the infection site by means of chemotaxis.

Each neutrophil can hold upto 15-20 micro-organism at a time.

Start destroying the bacteria by means of phagocytosis.

72

• Coarse large granules that stain pink with eosin.

• Nucleus is bilobed (spectacle shaped)• Diameter=10-14μ

Eosinophils

H&E staining

GEIMSA staining

Elect. Micros. view

Provides defense against parasitic infections and allergic conditions.

They are responsible for detoxification, disintegration and removal of foreign proteins.

73

Mechanism of action:

Attack the invading organism by secreting cytotoxic substances.

These substances are lethal and destroy the parasites

They are eosinophil peroxidase,major basic protein (MBP), eosinophil derived neurotoxin and interleukin 4 and 5

74

• Also have Coarse large granules • Stain purple blue with methylene blue

• Nucleus is bilobed

• Diameter = 8-10μ

Basophils:

Elect. Micros. view

These play an important role in healing process and acute hypersensitivity reactions.

75

Mechanism of action:

Basophils execute functions by releasing important substances from their granules such as heparin and histamines,proteases and myeloperoxidases and interleukin-4.

76

• Largest WBC’s, Diameter = 14-18μ • Cytoplasm clear without granules

• Nucleus is oval, bean shaped or kidney shaped

• Nucleus is in the center pushed to one side

• Large amount of cytoplasm is seen

Monocytes

77

These are the largest cells among the WBC’s.

Like neutrophils they are motile and phagocytic in nature.

They are the precursors of the tissue macrophages.

Matured monocytes stay in blood for few hours.

After which they enter the tissues and become tissue macrophages.

78

These act by secreting substances like interleukin-1, colony stimulating factor and platlet activating factor.

Mechanism of action:

79

Lymphoytes:

• Cytoplasm clear without granules

• Nucleus is oval, bean shaped or kidney shaped and occupies whole of the cytoplasm.

• only a rim of cytoplasm may be seen

80

•Depending upon the size, they are divided into two types:

•Large lymphocyte: younger cells diameter= 10-12μ

•Small lymphocytes: older cells with a diameter= 7-10μ

•depending upon functions they are further divided into: T-lymphocyte- concerned with cellular B-lymphocytes-concerned with humoral immunity• These are responsible for development of immunity.

81

Physiological Varriations Age: In infants and children, WBC count is more-20,000/cummIn adults it is 4000 to 11000/cumm of blood

Sex: Slightly more in males than in females.

Exercise: Increases slightly

Emotional conditions: Increases slightly

Pregnancy: Increases

82

Pathological Variations Leucocytosis: occurs in following pathological conditions Infections Allergy Common cold Tuberculosis Glandular Leukopenia: occurs in following pathological conditions Anaphylactic shock Cirrhosis of liver Disorders of spleen Viral infections Pernicious anaemia

83

Leukemia: Characterized by abnormal and uncontrolled increase.

Increase in WBC’s, above 1000,000/cumm.

Also called blood cancer.

All the WBC’s may not increase at one time.

Leucocytosis occurs because of increase in any one of the WBC’s

84

85

86

Properties of WBC’s Diapedesis: property of WBC’s to squeeze through narrow blood vessels.

Ameboid movement: neutrophils, monocytes and lymphocytes show amebic movement characterized by protrusion of the cytoplasm and change in shape.

Chemotaxis : Attraction of WBC’s towards the injured tissues by chemical substances released at the site of injury

Phagocytosis : Neutrophils and Monocytes engulf the foreign bodies by means of phagocytosis.

87

Functions of WBC’s

WBC’s generally play an important role in the defense mechanism of the body.

In the defense mechanism each type of WBC’s act in a different way

88

NATURAL KILLER CELLS•They have large granular cell with independent nucleus.•Considered as third type of lymphocyte.•Kills invading organism or virus and virus infected cell, malignant cell•First line of defense against viruses.

89

PLATELETS.Thrombocytes (platelets) are fragments of megakaryocytes(red bone marrow) Cytoplasm is present but do not have a nucleusHave cell membrane with microtubules below it. 2–3 µm in diameter Normal range : 150,000 to 400,000 per cmm Circulation in blood – 8-12 days

90

Platelets- functions

•The main function of platelets is the maintenance of hemostasis. •Repair of ruptured blood vessels• Clot retraction• Procoagulant• Inflammation•Role in defense mechanism.

91

Thrombopoiesis Platelets are produced in bone marrow, by budding off from megakaryocytes. Megakaryocyte and platelet production is regulated by thrombopoietin, a hormone usually produced by the liver and kidneys Each megakaryocyte produces between 5,000 and 10,000 platelets. Reserve platelets are stored in the spleen, and are released when needed by sympathetically induced splenic contraction. Old platelets are destroyed by phagocytosis in the spleen and by Kupffer cells in the liver

92

MAST CELLSThese are large tissue cell resembling the basophil.Present in bone marrow and around cutaneous blood vessels.It do not enter blood circulationThey play important role in producing hypersensitivity reaction like allergy and anaphylaxis.It secretes histamin serotonin and hydrolytic enzymes.

93

COMPLETE BLOOD COUNT(CBC)• The complete blood count, or CBC, lists a number of many important values.

Typically, it includes the following:• White blood cell count (WBC or leukocyte count)• WBC differential count• Red blood cell count (RBC or erythrocyte count)• Hematocrit(Hct)• Hemoglobin(Hbg)• Mean corpuscular volume (MCV)• Mean corpuscular hemoglobin (MCH)• Mean corpuscular hemoglobin concentration(MCHC)• Red cell distribution width (RDW)• Platelet count• Mean Platelet Volume (MPV

94

•WBC (white blood cell) (TLC)4,300 and 11,000 cells per cubic millimeter (cmm).•RBC (red blood cell) ranges between 4.2 to 5.9 million cells per cmm.•Hemoglobin (Hbg) 13 to 18 grams per deciliter (one-hundredth of a liter) for men and 12 to 16 grams per deciliter for women.•Hematocrit (Hct) 45%-52% for men and 37%-48% for women.•Mean corpuscular volume (MCV) ranges between 80 to 100 femtoliters (a fraction of one-millionth of a liter).•Mean corpuscular hemoglobin (MCH) ranges between 27 to 32 picograms (a small fraction of a gram).•Mean corpuscular hemoglobin concentration (MCHC) ranges between 32%-36%.•Red cell distribution width (RDW) ranges between 11 to 15.•Platelet count ranges between 150,000 to 400,000 per cmm.•Mean platelet volume (MPV). The normal range is between 6 to 12 femtoliters.

95

THANK YOU