BLOOD DR/ BASSAM AHMED

BLOOD
-Examination of the blood:
-Staining: with Neutral stain e.g. leishman's stain: formed of mixture of:
a) red acidic stain (eosin)
b) Blue basic stain (methylene blue) dissolved in methyl alcohol (fixative).

Red Blood Corpuscles (RBCs) (Erythrocytes)

-Shape:
-In top view: rounded. -In side view: biconcave, to increase the surface area for gas exchange.

-Size:
-6-9 µm in diameter with as average 7.5 µm.

-LM:
-In unstained blood film: RBCs appear colored due to Hb.
-In blood film stained: RBCs are rounded, non-nucleated & acidophilic (Hb is a basic protein), with
pale center (about 1/3 of the diameter of RBCs), Normochromic RBCs.

*NOTE:
''Erythrocyte Sedimentation Rate'' (ESR): long chains of RBCs sediment more easily
which increases with inflammation.

-EM:
-Have no nucleus or organelles, filled with hemoglobin which appears electron-dense & homogenous.
-The cell membrane is flexible.
-Their cytoskeleton [actin and spectrin] keep their shape and stability of the membrane.

-Life span: is about 120 days.

-Fate:-Old RBCs are phagocytosed by macrophages in the liver, bone marrow and spleen.

-Osmotic Fragility of RBCs:

-RBCs maintain their normal shape in plasma [isotonic solution = the O.P of 0.9% saline].
.Crenation: if placed in hypertonic solution, they shrink and show notches.
.hemolysis: if placed in hypotonic solution, they swell, burst and leak hemoglobin,
the remaining cell membrane is called cell ghost.

-Number of RBCs:
.Average number of RBCs is 5 million /mm3.
.In normal: a)adult male, ranges from 5 – 5.5 miilioin /mm3.
b)adult females, ranges from 4.5 -5 miilion /mm3??
-Due to the stimulatory effects of male hormones on the bone marrow.

-Note: RBC count is highest in the newborns and decreases gradually.

 BLOOD DR/ BASSAM AHMED
-Abnormalities in number: Essay
(I)Anemia: decreased number of RBCs below 4 million/mm3 (oligocythemia) and/or
decreased hemoglobin concentration.
*Pernicious anemia: (Vitamin B12 deficiency anemia) due to failed production of intrinsic factor by
stomach.

*Sickle cell anemia: Formation of abnormal rigid type of Hb Called (Hb S) that accumulates at one
side of the cell giving acrescent shape (Sickle RBCs).

*Aplastic anemia: destruction of the bone marrow e.g. by irradiation or chemotherapy leading to
pancytopenia (decreased count of all blood cells).

(II)Polycythemia:
-Increased number of RBCs above 6 million/mm3 , due to hypoxia that stimulates the bone marrow to
produce more RBCs.
-It may be: a)Physiological: e.g. high altitudes, muscular exercise and in newborns.
b)Pathological: e.g. chronic lung and heart diseases

*Adaptation Of RBCs To Perform Their Functions: Essay

Factor Mode of adaptation
(I) Plasmalemma 1- flexible to be squeezed inside narrow capillaries.
2- IT is lipoprotein, [highly selective] for gas exchange.
(II) Shape 1- Biconcave increasing surface area for gas exchange.
2- have rounded edges for easy passage in branched vasselse.
(III) content 1- No nuclei or organelles, so: don't divide & -give more space for Hb.
[33% H b, 66% H2o & 2-Contain enzymes as: a) Hb Reductase (combine with O2)
1% enzymes]. b) carbonic anhydrase (carry CO2).

PLATELETS [THROMBOCYTES]
-Origin: develop from megakaryocyte. -No: 200,000 – 400,000/ mm3.
-L.M. Essay
oval, non-nucleated fragments which have 2 zones:
-Outer pale basophilic (clear) peripheral zone = Hyalomere.
-Central dark granular zone = granulomere

-E.M & related functions: Essay

-The cell membrane have thick cell coat that help platelets aggregation [platelet plug] to stop bleeding.

1) Hyalomere contains:
1- Cytoskeleton:
a- Microtubules: maintain cell shape.
b- Actin microfilaments: help activated platelets to change their shape & aid clot retraction.
2- Membranous Channels:
a- open canalicular system: invaginations of C.M to release serotonin: vasoconstriction of injured vessel.
b- dense tubular system: store for Ca**
2) Granulomere contains:
-Few mitochondria, ribosomes, glycogen (energy) & 3 types of granules:
1. [specific] granules: contain clotting factors & growth factors & Ca++.
 BLOOD DR/ BASSAM AHMED
2. Delta [Dense] granules: contain serotonin , ATP & ADP
3. Lambda granules: lysosomes for clot removal after healing of the vessel.

*Purpura = thrombocytopenia:
↓↓N o of platelets < 50.00/mm3 due to: a)↓↓production [b.m. depression]
b)↑↑ breakdown [autoimmune disease]
-Characterized by: -Prolonged bleeding time -Excessive bleeding after minor trauma

-Comparison Between Erythrocytes and Leucocytes: Essay

--------------- Erythrocytes Leucocytes
Type One type Five types
Content Not true cells (No nuclei nor organelles) True cells with nuclei and organelles
Number 4.5-5.5 millions/mm3 in males and 4.000- 11.000/mm2
3
4-5 millions/mm in females
Size 6-9 (7.5 µm) 6-20 µm
Life span 120 days Days to years
Osmotic fragility Easily hemolysed Not easily hemolysed
Origin & Bone marrow Bone marrow and lymphoid tissue
maturation
Shape Biconcave discs Spherical
Function Gas exchange Defense
Motility Non motile Motile

-White Blood Cells (WBCs) (Leucocyte)

1) Granular leucocytes: Essay
1-neutrophils
-called: polymorphonuclear leucocytes = P.M.Ns.
-% : 60-70%

-L.M
-Nucleus: single but segmented 2-5 lobes connected by thin chromatin thread, dark &
many shapes (P .M.N.), Barr body can be seen in 3-6% ♀ cells.
-Cytoplasm: specific granules: numerous fine, pale granules.
-E.M:
-N: peripheral heterochromatin & small central euchromatin.
-C: Have pseudopodia -Few organelles (mitochondria, few rER ) -rich in glycogen (energy), small GA

-Granules: 2 types:

1) Azurophil (1ry) 2) Specific (2 ry)

●large, few, dark: considered as 1ry ●Small, many & paly, contain:
-Lysosomes: contain: 1. Collagenase. 2. Phagocytin.
1. Myelo-peroxidase 3. Lysozyme. 4. Lactoferrin.
2. Defense against bacteria, Fungi and viruses.

Functions: 1st line of defense (non-specific)

●Bacterial toxins attract neutrophils which leave the blood by migrating between endothelial
cells of blood vessels to c.t by a process called ''diapedesis'' .
 BLOOD DR/ BASSAM AHMED
1) phagocytose bacteria:
First: specific granules release their contents into the phagocytic vesicle & CT:
-Lactoferrin: binds to iron (essential for bacterial growth) (Bacteriostatic)
-Phagocytin: kill bacteria. (bactericidal)
-Lysozyme: destroys collagen so facilitates movement of neutrophils
Second: azurophil granules release their
-Enzymes: myeloperoxidase kill bacteria & other enzymes cause their lysis.

2) Dead neutrophils from pus cells.

3) pus ↑↑ body temperature: (pyrogenic) stimulates the heat regulating center in the brain.
4) Attract monocytes to site of infection: to clean it
5) Secrete trephone substance: help healing of wounds.

-Increased %: Neutrophilia (>75%): due to acute pyogenic infection e.g. acute tonsillitis, appendicitis &
abscess.
-Decreased % = Neutropenia(<60% ): Typhoid fever (commonest cause), TB, Viral infection as influenza

2. Eosinophils 3. Basophils
%: 1-4 % -Diameter -10-14 µm -Life span: few days -%: 0-1 % -Diameter: 10-12 µm

L.M. N. Horse-shoe shaped (biloed) connected N. Irregular, segmented pale and s-shaped.
By thick chromatin thread
C. Contain large acidophilic C. Coarse basophilic granules
specific granules Metachromatically stained with toluidine
blue →purple (due to presences of heparin).
E.M. N. Peripheral heterochromatin with more N. has minimal peripheral heterochromatin
central euchromatin than in neutrophil. with more central euchromatin
C. -Few organelles C. -Few organells
-2 types of granules: -2 types of granule:
●Azurophil: oval containing histaminase, Azurophil: Small = lysosomes
sulphatase & eosinophil derived neurotoxin Specific: Large, rounded, electron dense &
-contain: -Histamine -Heparin -Leukotriens.
-eosinophil chemotactic factor
-Cell membrane shows receptors for lg E.
-Function: -Functions
1. Termination of allergic reaction: 1.Secrete Heparin (prevents clotting & promotes
a-secrete Histaminase & sulphatase to destroy allergy)
histamine & heparine & end allergy 2.Secrete Histamine → vasodilatation → sudden ↓↓
b- Phagocytose the antigen-Ab. Complexes in blood Pressure → anaphylaxis
2. Defend against parasites: 3.Attract eosinophils to site of allergy by Eosinophil
a- Cytotoxic effect: from pores in their bodies. chemotactic factor
b- Neurotoxins: Nervous dysfunction of parasites. 4.Leukotrienes: bronchospasm →bronchial asthma.
5. Limited phagocytic power

↑↑№ = Eosinophilia> 5% ↑↑ № = Basophilia: more than 2%

1. Allergic distance: Urticaria, eczema & bronch asthma 1. Allergic diseases. 2. Parasitic diseases.
2. Parasitic diseases: Bilharziasis. 3. Viral infection (chicken pox). 4. Liver cirrhosis.

↓↓ № = Eosinopenia< 1% ------------------------------------
Treatment with cortisone (inhibit release from B.M)
 BLOOD DR/ BASSAM AHMED

Basophil Mast cells

Life span Few days Longer (weeks to months)
Size 10- 12 µm 20- 30 µm
The Nucleus Segmented Round
Phagocytosis Slight No
Granules Metachromatically stained due to Same basophil
(Histamine & heparin)
Surface Antibody lg E Same basophil
receptors

2- Non-Granular leucocytes: Essay

I- Monocytes:
%: 3-8% Diameter: 13 - 20µm Life span: 3 days in blood & 3 months in c.t.
L.M.: Nucleus: Large, eccentric, kidney-shaped, pale with 1 or 2 nucleoli.
Cytoplasm: non-granular, non-clear pale basophilic -Has frosted glass appearance due to lysosomes
E.M:
N.: Euchnomatic with clear nucleolus
C.: have pseudopodia, mitochondria, r-ER, well develop G.A, many lysosomes

Functions: Highly phagocytic cells:

1. Remain in blood for 3 days then
2. Enter c.t. & change into macrophages to phagocytose bacteria & dedebris.
3. Antigen presenting cells.
4. The precursors of all phagocytic cells: e.g. dust cells of lungs, kupffercells of liver, osteoclasts of bone

↑↑№: Monocytoses: > 8% due to chronic infections: T.B., syphilis & glandular fever, Monocytic leukemia
↓↓№: Monocytopenia: less tan 3% pancytopenia.

II- lymphocytes
-20- 30% ●2nd most common cell of WBCs ●2nd line of defense
●Immune competent cells: have surface markers (receptors) for antigens.

↑↑№: lymphocytosis:> 30%:

a) Physiological in children b) Chronic infections e.g. T.B &whooping cough c) Leukemia
↓↓№: lymphocytopenia (pancytopenia)

-Two types according to activity: Essay

Small Large
% -15 -20% of WBCs -5 – 10%
Diameter : -6 -8 µm -10 -15 µm
L.M. : -N.: dark & filling the cell N. large, indented, pale with clear nucleolus
C. abundant, deeply basophilic
-C.: little, pale basophilic rim around
the nucleus N. Euchromatic with prominent nucleolus
E.M. -N. heterochromatic C. more organelles: mit., ribosomes, rER, G.A.
-C. few mit., small GA, many free + lysosomes
ribosomes, two centrioles + small
azurophil granules
 BLOOD DR/ BASSAM AHMED

-Classification of lymphocytes: have similar L.M. or E.M. they differ only in the surface receptors
Three types according to function: Essay
T- lymphocytes B- Lymphocytes Natural killer (NK cells)
% 60 – 80% 20 – 30% 5 – 10 %
Life span Years Few days → few months Years
Maturation Site Stem cells in b.m. migrate to -Burse of fabricus in birds Develop from precursors of T
thymus gland, differentiate & -Bone marrow in & B cell but do not mature in
acquire receptors (thymic- Mammals where they thymus
education) Develop receptors
Surface markers T –cell receptors (TCR) e.g. Receptors for: - CD16 & others.
(receptors) CD4 & CD8 ptn Ig M, lg D. - have neither T nor B-
cell receptors
Function Cell-Mediated immunity Humoral immunity Innate immune response.
(early) without stimulation by
T-helper cells
-similar to cytotoxic T-cell
- secretion of interferon
(antiviral)

T- lymphocyte
-Function (Types): Essay
1- Cytotoxic CD8 T-cells:
-secrete perforins that from pores in the cell membrane of virally infected cells directly killing them.

2-Helper CD4 T-cells: Activate B-cell during humoral immunity

3-Regulatory (Suppressor) T-cells:
1-Maintain unresponsive to self-antigen. 2-Suppress XSS immune responce

4-Memory T-cells: -rapid response of T-cells on re exposure to the same Ag. (2ry immune response)

B-Lymphocyte
-Functions: -Humoral Immunity: Essay
● when exposed to specific antigen, become activated by helper T-cell to:
1- plasmablasts then plasma cells that produce antibodies.
2-Give B-memory cells: responsible for the rapid response on the 2nd exposure to the same Ag.

-Note: HIV human immunodeficiency virus that causes AIDs destroys the T-helper cells →↓↓ immunity
& the patient becomes susceptible to infections
 BLOOD DR/ BASSAM AHMED
Leukocytic count
I-Total leucocytic count:
-The total No of WBCs/mm3 of blood = 4.000 to 11.000/mm3.
-Counted by → (1) Haemocytometer. (2) Electronic counting instrument.

-Abnormalities in total count: Essay

Leukocytosis: ↑↑ total No. of WBCs > 11,000/mm3.
1.physiological: (transient): during pregnancy, labor, cold bath & exercise
2.Pathological: due to infections either acute or chronic
Leukopenia: ↓↓No of WBCs <4,000/mm3. Causes:
-X-ray - irradiation -Excessive use of antibiotics e.g. chloramphenicol
-Typhoid fever -Influenza virus

II- Differential leucocytic count:

-The % of each type of WBCs to the total No. -Count in a blood film stained with leishman.

Haemopoiesis
-Formation of blood cells in: a) bone marrow b) lymphatic organs e.g. thymus
-Bone marrow = (myeloid tissue)
Red Bone Marrow (active) Yellow Bone Marrow (inactive)
Present in most bones of children In shafts of long bones of adults
In adults : flat, short & irregular bones A store for fat can change into active on need

-Structure of red bone marrow: Essay

1)Stroma:
-Fixed cells: reticular cells, together with reticular fibres from network in the background, fibroblasts, UMC,
pericytes, osteogenic cells, & fat cells (the largest cells in bone marrow).
-Blood sinusoids: Wide irregular blood capillaries lined with endothelial cells with non-continuous basement
membrane through their pores blood cells leave to the blood.

2) Free cells:
● The ratio of immature WBCs to immature RBCs=5:1 as the life span of most WBCs is shorter than that of RBCs

*Some Notes: Essay

- Reticulocyte:
-The cell become smaller - immature RBCs but slightly larger.
-Cytoplasm: acidophilic (Hb) with remnants of ribosomes & polysomes that form reticulum.
-In peripheral blood: its % doesn't exceed (1%)
-Their % increase in case of haemorrhage or destruction of RBCs.
-Can be stained by supravital stain = Brilliant Cresyl Blue.

- Megakaryocyte: very large

-Have single multilobed dark N and basophilic cytoplasm.
-Invaginations of cell membrane form demarcation channels dividing the cytoplasm into
fragments that shed as platelets.
-it extends pseudopodia (platelet ribbons) form which platelets detach.