Hema Lec Trans Prelim

OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024

HEMATOLOGY 1
MR. ROJOHN SONNY C. CRUZ, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: TUASON, MLYL
COURSE OUTLINE: PRELIMS

1. Basic Hematological Methods of Examination
2. Quality Assurance Program
3. Collection of Blood Specimen
4. Hematopoiesis
5. Lineage Specific Hematopoiesis
6. Red Blood Cell
REFERENCE BOOKS
th
Rodak’s Hematology 5 Edition
th
Rodak’s Hematology 6 Edition
INTRODUCTION TO
HEMATOLOGY
HEMATOLOGY
Greek Word: Haima-Blood
(heme - blood; logos - study) STUDY OF BLOOD
- primarily deals with the cellular elements of the peripheral

- blood and bone marrow.
- includes morphologic appearance, function, and disease of
- blood and blood forming organs.
HEMATOLOGY
Hematology - study of the clinical, morphologic and

laboratory disorders of the blood and other blood-forming
organs
BLOOD - red liquid that circulates in the arteries and veins of

humans and other vertebrate animals that carries oxygen
and nutrients inside the body.
Total blood volume in an adult is 5 to 6 liters or 7 to 8% of

the total body weight
The clinical hematology section perform scientific

analysis of blood
 Fluid portion of blood
 Plasma
 Serum
 Formed elements of blood
 Blood cells
 Coagulation Factors
 Protein components
HEMATOLOGY 1
CHARACTERISTICS OF BLOOD
 COLOR
Arterial blood: bright scarlet red WHOLE BLOOD COMPOSITION
Venous blood: dark red Whole blood includes erythrocytes, leukocytes, platelets, and
 VISCOSITY - thick and sticky fluid that normally flows with plasma. When a specimen is centrifuged, leukocytes and
difficulty platelets make up the buffy coat (small white layer of cells
 SPECIFIC GRAVITY - 1.055 to 1.065 lying between the packed red blood cells and the plasma).
 pH - 7.35 to 7.45
PLASMA VS. SERUM
Plasma is the liquid portion of unclotted blood. Serum is the
COLOR OF BLOOD fluid that remains after coagulation has occurred and a clot
has formed period.
a. Plasma is composed of 90% water and contains proteins,
enzymes, hormones, lipids, and salts.
b. Plasma normally appears hazy and pale yellow (contains
all coagulation proteins), and serum normally appears clear
and straw colored (lacks fibrinogen group coagulation
proteins).
FUNCTIONS OF BLOOD
I. METABOLIC FUNCTIONS:
1. RESPIRATION
- transport of oxygen from the lungs to the tissues and
carbon dioxide from the tissues to the lungs
2. NUTRITION
- transport of absorbed food materials
3. EXCRETION
- transport of metabolic waste to the kidneys, lungs, skin and
intestines for removal
4. Regulation of water balance through the effects of blood in
the exchange of water between circulating fluid and tissue
fluid.
5. Regulation of body temperature by distribution of body
heat
6. Transportation of hormones from origin to target cells
7. Maintenance of normal acid-base balance in the body
HEMATOLOGY 1
II. DEFENSIVE FUNCTIONS HANDWASHING

1. Production of immune globulins Hands must be washed:
2. Function as phagocyte 1 Whenever there is visible contamination with blood or body
• Neutrophils fluids
• Eosinophils 2. After completion of work
• Monocytes 3. After gloves are removed and between glove changes
4. Before leaving the laboratory
ROUTINE PROCEDURES 5. Before and after eating and drinking, smoking, applying
 Complete Blood Count - CBC cosmetics or lip balm, changing a contact lens, and using the
• Hemoglobin lavatory
• Hematocrit 6. Before and after all other activities that entail hand contact
• WBC Count with mucous membranes, eyes, or breaks in skin
• RBC Count
• Smear Preparation Applicable safety practices required by the OSHA
• Differential Count standard in hematology laboratory
• Platelet Count 1. Handwashing
 Red Blood Cell indices  Wet hands and wrists thoroughly under running water.
 ESR - erythrocyte sedimentation rate  Apply germicidal soap and rub hands vigorously for at
 Reticulocyte count least 15/20 seconds, including between the fingers and
 Peripheral blood smear – morphology around and over the fingernails.
 Rinse hands thoroughly under running water in a
downward flow from wrist to fingertips.
 Dry hands with a paper towel. Use the paper towel to
turn off the faucet handles.
2. Eating, drinking, smoking, and applying cosmetics or lip
balm must be prohibited in the laboratory work area.
3. Hands, pends, and other fomites must be kept away from
the mouth and all mucous membranes.
4. Food and drinks, including oral medications and tolerance
testing beverages, must NOT be kept in the same
refrigerator as the chemical reagents or specimen.
5. Mouth pipetting must be prohibited
6. Needles and other sharp objects contaminated with blood
and other potentially infectious materials should not be
manipulated in any way (resheating, bending, clipping, or
removing the sharp object)
Most Common Technical Hematologic Procedure ** Resheating or recapping is permitted only when there are
Includes: no other alternatives. The other one-handed method is often
- Principles and operation of laboratory equipment used.
- Procedure in blood collection 7. Contaminated sharps must be placed in a puncture-
- Quality control resistant container that is appropriately labeled with the
- Methods of evaluation universal biohazard symbol and that is leak proof.
- Determination of normal testing range 8. Procedures manipulating any form of liquid should be
performed in a way that prevents splashing, spraying, or
production of droplets in the specimen. It can be performed
GENERAL METHODS IN EXAMINING BLOOD behind a barrier such as plastic shield or safety goggles
should be worn.
1.0 Fresh Preparation 9. Wearing of Personal Protective Equipment
1.1 Poikilocytosis  Lab coat
1.2 Abnormal inclusions  Safety Goggles
1.3 Extracellular parasites  Gloves
2.0 Phase Microscopy  Masks
3.0 Electron Microscopy  Hair net
4.0 Fluorescent Microscopy 10. Phlebotomy trays should be appropriately labeled to
indicate potentially infectious materials. Specimens should
be placed into a secondary container, such as a resealable
biohazard-labeled bag.
LABORATORY SAFETY STANDARDS IN 11. Proper containers for pneumatic tube system of
HEMATOLOGY specimen transport. Primary containment and Secondary
containment. Foam to prevent shifting and as a shock
Occupational Safety and Health Administration (OSHA) absorber.
▪ the rule that specifies standard precautions to protect 12. Equipment or machine that have been contaminated
laboratory workers and other healthcare professionals before reusing or repairing.
became effective on March 6, 1992.
▪ Universal precautions - Standard precautions
"All blood, body fluids, and unfixed tissues are to be
handled as though they were potentially infectious"
HEMATOLOGY 1
Laboratory Safety
• Housekeeping
• Laundry
• Hepatitis B vaccination
• Training and documentation
• Regulated Medical Waste Management
Occupational Hazards
• Fire Hazards
• Chemical Hazards
• Electrical Hazards
• Needle Puncture
HEMATOLOGY 1
BASIC HEMATOLOGICAL QUALITY ASSURANCE PROGRAM

METHODS OF EXAMINATION QA is the sum of all those activities in which the laboratory is
engaged to ensure that information generated by the
1. Hematology test laboratory is correct.
CBC – a basic screening test and one of the most frequently
ordered laboratory procedure. Includes all aspects of laboratory activities that affect the
- Hgb, Hct, RBC and WBC count, Differential count results produced from the choice of method, to the handling
of specimen and reporting results.
WHITE BLOOD CELL COUNT (LEUKOCYTE COUNT)
- Test that measures the number of WBC in your body The real purpose is to determine how correct/incorrect the
- Is often included in CBC result is.
- A WBC count detects hidden infections
- Helps doctors monitor the effectiveness of chemotherapy or 3 MAJOR ACTIVITIES
radiation Preventive – activities done in prior to the examination of the
- Treatment in cancer patients
specimen or sample that is intended to establish system
conducive to accuracy testing.
RBC COUNT Assessment – activities done during testing to determine
Blood test used to find out how many RBCs you have whether the test systems are performing correctly.
Also known as Erythrocyte count Corrective – done when error is detected to correct the
Important because RBCs contain Hgb which carries oxygen system.
to your body tissues - Recalibration of the instrument/machine.
* Increase RBC – Polycythemia Vera
* Decrease RBC – Anemia
QA IN HEMATOLOGY LABORATORY
HGB DETERMINATION - intended to ensure the reliability of the laboratory test.
Measurement of the concentration of Hgb in the blood - objective is to achieve precision and accuracy.
Protein in RBC that carries oxygen to your body
Used also to rule out Anemia Accuracy is the closeness to the estimated value to the true
mean.
DIFFERENTIAL COUNT Precision is the reproducibility of a result, whether
- WBC differential count accurate or inaccurate.
- Percentage of each type of WBC
- Can detect immature WBC and abnormalities, both of BLOOD COLLECTION
which are signs of potential issues.
- RBC and platelets can be seen as well 1. SKIN PUNCTURE
- Blood sample collected is called peripheral blood
Hematocrit instead of capillary blood.
Volume of packed RBC that occupies a given volume of - Sample is a mixture of capillary, venous, and arterial
whole blood. blood with interstitial (tissue) fluid and intracellular fluid.
Reticulocyte count Sites of skin puncture

- Use to assess the erythropoietic activity of the bone marrow - finger (middle or ring)
- whole blood, anticoagulated with EDTA is stained with a - earlobe (less free nerve endings, less pain and less tissue
supravital stain such as new methylene blue or brilliant cresyl juice)
blue. <1 yr old – lateral portion of the plantar surface of the
heel/toe.
Erythrocyte Sedimentation Rate (ESR)
Rate at which red blood cells sediment in a period of one
hour.
- Common hematology test and nonspecific measure of
inflammation
HEMATOLOGY 1
METHOD OF COLLECTION
1. Syringe method
Order of Draw - Syringe Method
 Blood culture
 Other sterile tubes
 Light blue
 Serum sterile tubes
 Green
 Lavender
 Gray
 Yellow
 Red
Methods of Skin Puncture

 Heel Stick Procedure
 Finger Puncture – 3rd or 4th finger of non- dominant hand
 Earlobe Puncture – abnormalities in WBC and histiocytes
Sites to avoid ETS METHOD (EVACUATED TUBE SYSTEM)

 inflamed and pallor areas
 cold and cyanotic areas
 congested and edematous areas
 scarred and heavily calloused areas
Reasons for capillary puncture in infants and very young

children
 Advantages of skin puncture using earlobe
 Disadvantages of skin puncture
 Order of draw for skin puncture
2. VENIPUNCTURE
- manner of inserting a needle attached to a syringe to a
palpable vein to collect blood for laboratory testing. WINGED BLOOD COLLECTION SET
- blood sample collected: VENOUS BLOOD
- most widely used blood sample in all laboratory tests
3 factors involved in a good venipuncture

 Phlebotomist
 Patient and his/her vein
 The equipment needed
Key notes to remember on Venipuncture

 Angle of needle insertion
o Routine venipuncture – 45 degrees (30-45 degrees)
 Donor bleeding – 16gauge, skin to vein
 Needle should be bevel up
 Length of needle - 1 to 1.5 inches
 Tourniquet – 1 min (reapply within 2 minutes)
 Blood pressure cup – 40 to 60 mmHg
 Number of attempts – maximum of 2
 Position of the patient
HEMATOLOGY 1
Sites of venipuncture 3 y/o to adult life

In newborn infants up to 18 months old - wrist vein
- External jugular veinp - dorsal vein of the hand
- Temporal vein (scalp vein) - dorsal vein of the ankle
- Superior longitudinal sinus
IN OLDER CHILDREN (18 MONTHS TO 3 Y/O)

- Femoral vein
- Long saphenous vein
- Popliteal vein
- Ankle vein
VEINS OF THE ANTECUBITAL FOSSA

Two patterns of vein
 “H” pattern
o Median capital vein
o Cephalic vein
o Basilica vein4
 “M” pattern
o Median
o Accessory cephalic vein
o Basilic vein
ORDER OF DRAW FOR VENIPUNCTURE

(You Better Remember Girls Loves Gays)
 Blood culture (yellow stopper)
 Coagulation tube (blue)
 Serum tubes with or without activator
 Heparin (green)
 EDTA (lavender)
 Sodium fluoride tube with or without EDTA or oxalate (gray
stopper)
Advantages of Evacuated Tube Method

 Direct
Complications
Sometimes, vacuum on the tube is not enough which can
affect the testing
HEMATOLOGY 1
MOST COMMON ANTICOAGULANT USED IN 3 forms of oxalate:

HEMATOLOGY  Double oxalate
1. EDTA/Versene/Sequestrene  Lithium Oxalate
 Most commonly used  Sodium Oxalate
 MOA: bind to non-ionized form of Calcium then chelates
Calcium Double Oxalate
 salts of Ammonium and Potassium
3 forms:  when used separately:
 Dipotassium EDTA o Ammonium oxalate (RBC Swells)
 Disodium EDTA o Potassium oxalate (RBC shrinks)
 Tripotassium EDTA  Also known as “Balance Oxalate”
o Recommended amount: 1.2 mg/dl of blood  Uses: all RBC valuation tests, since there is no effect on
o Used for: CBC, ESR, Platelet count and PBS RBC
o Can be used for modified Westergren (ESR)
 2 ml EDTA blood + 0.5 ml of 3.8% of 0.129 M Na DISADVANATGES:
Citrate  Not recommended for blood transfusions
 2 ml of EDTA blood + 0.5 ml NSS  Causes agglutination or clumping of WBCs and platelets
DISADVANTAGES: Not recommended for peripheral blood smear

 Not recommended for coagulation and platelet satellitism preparation
 Preparation of blood smear from old EDTA sample
changes the morphology of the red cell HEMATOPOIESIS
2. Citrate A continuous, regulated process of blood cell production that
 Most common and preferred anticoagulant for coagulation includes cell renewal, proliferation, differentiation, and
studies maturation.
 MOA: binds calcium
 Contained in light blue top Evacuated tube These processes result in the formation, development, and
 Anticoagulant buffer: 3.8% / 0.109 M Na Citrate specialization of all of the functional blood cells that are
released from the bone marrow to the circulation.
HEMATOPOIESIS
 Blood cell production
 Cellular proliferation
 Differentiation
 Morphogenesis
 Functional maturation
3. Heparin  Death
 Most commonly used for Osmotic Fragility Testing and
immunotyping STEM CELL THEORY
 MOA: binds thrombin A. Monophyletic theory - suggests that all blood cells are
derived from a single progenitor stem cell called a pluripotent
2 forms: hematopoietic stem cell. The monophyletic theory is the most
 Lithium Heparin widely accepted theory among experimental hematologists
 Sodium heparin today
 Uses: RBC count, haemoglobin, hematocrit, ESR and OFT
B. Polyphyletic theory - suggests that each of the blood cell
DISADVANTAGES: lineages is derived from its own unique stem cell.
 Not recommended for coagulation studies because it
affects all stages of blood coagulation
 Not for WBC
[FIGURE 1]
 Not recommended for blood smear preparation
 Most expensive
4. Oxalates
 MOA: binds Calcium
 Recommended concentration: 1-2 mg/ml of blood
HEMATOLOGY 1
STAGES OF HEMATOPOIESIS ADULT HEMOGLOBIN GLOBIN CHAIN

 Prenatal Hematopoiesis COMBINATION
Hb A 2 alpha + 2 beta
 Adult Hematopoiesis
Hb A2 2 alpha + 2 delta
PRENATAL HEMATOPOIESIS
ADULT HEMATOPOIESIS
1. MESOBLASTIC PERIOD
BONE MARROW - is the only site of erythropoiesis,
 YOLK SAC chief site of hematopoiesis in mesoblastic
myelopoiesis and thrombopoiesis
stage
 Nineteenth day of embryologic development after 1st few years of life - a delicate balance exists between
fertilization. developing bone marrow space and the developing infant's
 2nd week of fetal life, formation of blood islands in yolk sac need for blood cells and the liver or spleen remains available
(mesodermal extraembryonic layer), aggregation of because of its hematopoietic capability
primitive cells
 The primitive erythroblasts found in the yolk sac arise from 4th year of life - rate of bone marrow growth exceeds the
mesodermal cells, which initially line the cavity of the yolk need for blood cells therefore, active marrow sites are
sac. replaced with areas of fatty reserve and fat first develops in
 9th week of fetal life, development of primitive erythroblast the long bones
EMBRYONIC GLOBULIN CHAIN
HEMOGLOBIN COMBINATION 18th year of life - the only active hematopoietic sites are in
Gower I 2 zeta + 2 epsilon the pelvis, vertebrae, ribs, scapulae, sternum, skull and
Gower II 2 alpha + 2 epsilon proximal extremities of the long bones
Portland 2 zeta + 2 gamma
2. HEPATIC PERIOD
 Begins at 4 to 5 gestational weeks
 Starts on the 2nd month

 LIVER - CHIEF SITE OF HEMATOPOIESIS IN HEPATIC
STAGE; 3rd month of fetal life
Significant Contributions of Other Organs:

a. Thymus - T cell production
b. Kidney - B cell
c. Spleen B cell
 Erythropoiesis - until the end of normal gestation
(splenic)
 Myelopoiesis - but becomes minimal by the 5th month THE BONE MARROW
 Lymphopoiesis - lifetime  Develops in the embryo by the hollowing out of the skeletal
d. Lymph nodes - NRBC's, Granulocytes, Monocytes, bones forming a central cavity
Lymphocytes and Megakaryocytes  In this cavity develops a primitive, undifferentiated cell
FETAL HEMOGLOBIN GLOBIN CHAIN known as a hemocytoblast, or stem cell
COMBINATION  ALL blood formed elements ultimately develop from this
Hb F 2 alpha + 2 gamma undifferentiated precursor
3. MEDULLARY/MYELOID PERIOD
 Hematopoiesis begins in the developing bone marrow
cavity.
o Red bone marrow - CHIEF SITE OF HEMATOPOIESIS IN
MEDULLARY PHASE
o Starts on the 5th month of fetal life
 During this stage of development, the myeloid-to-erythroid
ratio approaches the adult level of 3:1 by 21 weeks of
gestation
o Development of cells increases at birth
o Hb A, - begins to appear and gradually increases in
concentration
 By the end of the sixth month, the bone marrow becomes
the primary site of hematopoiesis
HEMATOLOGY 1
 Bone marrow, one of the largest organs in the body, is RED MARROW - Site of blood cell development. It
defined as the tissue located within the cavities of the comprises approximately 50% of the marrow cavity space.
cortical bones The remaining 50% of the space is occupied by fat and is
 These cavities consist of trabecular bone resembling a known as yellow marrow. The ratio of red marrow to yellow
honeycomb. marrow is an indirect representation of marrow activity, and
 Normal bone marrow located within these cavities consists is expressed as marrow cellularity.
of two
TYPES OF MARROW:
1. RED MARROW, which is hematopoietically active marrow,

2. YELLOW MARROW, representing hematopoietically
inactive marrow composed primarily of adipocytes (fat cells).
MAJOR SITE OF HAEMATOPOIESIS

 It is found within the cavities of all bones and may be
present in two forms:
a. Yellow Marrow (composed mostly of fat (adipose
tissues))
b. Red Marrow (normally active in the production of most
types of leukocytes, erythrocytes and thrombocytes)
It represents approximately 3.5% - 6.0% of total body
weight.
 Average around 1500 g in adults.
 It is consists of hematopoietic cells (erythroid, myeloid,
lymphoid and megakaryocytes)
 Site of B cells to mature (production of RBC)
STRUCTURE AND DEVELOPMENT

Fat cell occupation of the bone marrow space
 Begins by 4 years of age BONE MARROW
 Occurs first at the diaphysis of long bones and slowly
 Full of developing precursor cells in all stages of
extends to the center of the bone
maturation
 By 18 years of age fatty replacement has limited active
 Released into blood at maturity
hematopoietic marrow to the following sites:
 ONLY mature cells are released normally
o Vertebrae
 First recognizable precursor in each cell line is a blast.
o Ribs
o Skull
o Sternum
o Proximal epiphyses of long bones
o Iliac crest of the pelvis
INDICATIONS FOR BONE MARROW STUDIES

1. Anemia, erythrocytosis, polycythemia
2. Leukopenia, leukocytosis, immature/abnormal cells in the
circulation
3. Thrombocytopenia, thrombocytosis
4. Malignant tumors
5. Infections
6. Diagnosis of hereditary or acquired storage of iron
HEMATOLOGY 1
BONE MARROW EXAMINATION

METHODS:
1. ASPIRATION - needle is inserted into the soft corner of
the bone and small quantity of bone marrow is aspirated,
using 30-35 ml. Syringe is used to aspirate 1-3 ml
SITES:
 Iliac crest - another common site for bone marrow
aspirate especially foe patients who are apprehensive
because he cannot see the procedure done.
 Sternum - usual site of puncture in adults, in the midline
of the bone between the second and third ribs. PREPARATION OF BONE MARROW FOR STUDY:
 Spinous process of the lumbar vertebrae  0.3 to 0.5 ml of specimen is obtained
 Tibia in children under 2 years of age.  As a rule marrow sample will be unavoidably diluted with
some peripheral blood.
2. BIOPSY- piece of the bone marrow is removed intact  Smears may be prepared as fast as possible before the
without disturbing the architecture of the bone specimen coagulates.
 Needle biopsy -done w/ a thick, hollow biopsy needle, and  Marrow aspirate should be placed in EDTA to inhibit
core sample of the bone marrow is taken clotting.
 Trephine biopsy-trephine is a small object/instrument used
to remove a circular section of tissue MARROW SMEARS WITHOUT ANTICOAGULANT:
 Surgical biopsy 1. WEDGE TYPE - prepared by placing a minute drop of
 Percutaneous biopsy specimen to slide and then spread it by using another slide.
2. PARTICLE SMEAR - it is done by placing several marrow
particles on the slide without fluid and it also spread in the
same manner as of the wedge type.
3. CRUSHED PARTICLE SMEAR - uses also the marrow
particles without the fluid portion but this time it is made by
gently squashing a particle between two slides or two cover
slips and then pulling the slides or cover slips apart in a
parallel position. This may lead to distortion of cells or
sometimes it results to a thick smear that is difficult to stain.
4. TOUCH PREPARATION - the result after an imprint of the
specimen has been left on a slide when this same specimen
has been transferred to a bottle with fixative like zenker that
may be used in cytology section.
MYELOID ERYTHROID (M:E RATIO)

 The two principal bone marrow lines which are the myeloid
erythroid can be expressed as M:E ratio.
 This ratio compares the number of developing
granulocytes (neutrophils, eosinophils, basophils) with that
of the relative number of erythrocyte precursors such as
erythroblasts.
 The expected marrow M:E ratio in adults ranges from 3:1
to 4:1. in infants may be higher.
HEMATOLOGY 1
Scenario M:E ratio Interpretation

(M:E ratio)
Infection 6:1 Increase
Leukemia 25:1 Increase
Myeloid 20:1 Increase
hyperplasia
Myeloid 3:20 Decrease
hypoplasia
Erythroid 1:20 Decrease
hyperplasia
B. MAST CELLS
Erythroid 5:1 Increase
hypoplasia 12-25 um, rounded or spindle shaped. Nucleus is round &
centrally located and purple. Cytoplasm is pink but w/ dark
purple to blue-black uniform spherical granules.
BONE MARROW DIFFERENTIAL
C. OSTEOBLASTS
- specialized cells which synthesize new bone matrix. Oval
cell, 30 um w/ small oval nucleus which is partially extruded
BONE MARROW DIFFERENTIAL – A procedure which
giving the cell "waterbug" or "comet" appearance. Cytoplasm
requires counting at least 500, and preferably 1000 cells be
which is blue is grainy.
counted for a marrow differential.
D. OSTEOCLASTS
- large multinucleated cells that are involved in bone
demineralization and resorption. It is likely that they are
formed from the fusion of circulating monocytes and
macrophages. Size is 100 um or greater which are irregularly
shaped and have ruffled cytoplasmic border. With variable
number of discrete nuclei and w/ cloudy light blue to light
pink cytoplasm.
NORMAL MARROW CELLS

A. MACROPHAGES
- most mature cell of the phagocytic system. Large, w/ a
diameter of 30 um, spreading, irregularly shaped cells &
pseudopods seen, w/ round or oval or elongated nucleus.
HEMATOLOGY 1
EXTRAMEDULLARY
HEMATOPOIESIS
EXTRAMEDULLARY HEMATOPOIESIS
1. Blood cell production in hematopoietic tissue other than
bone marrow.
2. Occurs when hyperplasia (increase in number of cells per
volume of tissue) of marrow cannot meet physiologic blood
needs of tissue.
3. Principally occurs in liver and spleen (just like in fetus)
lymph nodes and thymus
LIVER
Cellular Functions:
 Synthesizes various transport proteins
 Stores essential minerals and vitamins that are used in
DNA and RNA synthesis LYMPH NODES
 Conjugation of bilirubin hemoglobin degradation 
 Transports bilirubin to the small intestines for excretion  are located along lymphatic ducts and serve as central
 Kupffer cells - macrophages in liver cells collecting points for lymph fluid from adjacent tissues.
SPLEEN Lymph nodes act as lymphoid filters in the lymphatic
 Reticuloendothelial organ system. Lymph nodes respond to antigens introduced
 Primary functions - lymphopoiesis, phagocytosis -Located distally and routed to them by afferent lymphatics
in the left side of the abdomen just below the diaphragm
and behind the fundus of the stomach Layers
 Largest structure of the lymphoid system a. Cortex - contains macrophages, follicular dendritic cells,
 Site of extramedullary hematopoiesis naïve or resting B cells (in primary follicle), proliferating B
 Stores 30% of platelets (1/3 of total platelet) cells and plasma cells (in 2ndary follicles or Germinal
 Splenic tissue can be divided into two main types: Red centers)
pulp, White pulp b. Paracortex- contains mainly T cells
c. Medulla - contains T cells, B cells, plasma cells, and
1. WHITE PULP - contains lymphoid tissue macrophages
a. PALS (periarteriolar lymphoid sheaths) - Contains mainly
T cells THYMUS
b. Primary follicles - contains naïve B cells Thymus Gland
c. Marginal zone- contains dendritic cells - Responsible for normal development of some of the
lymphocytes
2. RED PULP - contains specialized macrophages for - Located in the neck
removal of the senescent RBCs: - Maximum development in childhood, atrophies with age
a. The red pulp makes up more than one half of the total
volume, and its function is to destroy senescent RBCs Bursa Fabricus
(Culling) or remove RBC inclusion (pitting) - Found in birds with possible analogous tissue in man.
b. CULLING - removal of mature RBC through phagocytosis - Responsible for normal antibody production
that leads to eventual degradation of cell organelles
c. PITTING - removal of cell inclusions.
LYMPH NODES
 Oval structures distributed throughout the body connected
by lymphatic vessels which carry a fluid called lymph.
 Act as filters to remove foreign blood contaminants.
Extremely important part of the body's infection defense
 Contain many phagocytic cells and lymphocytes
 Immature lymphocytes produced in the bone marrow
collect and mature in the tissues of the nodes
HEMATOLOGY 1
STEM CELL THEORY OF HEMATOPOIESIS
STEM CELL THEORY OF HEMATOPOIESIS

 All cells are derived from a pool of stem cells that are self
renewing
 Pluripotential & multipotential stem cells give rise to
committed stem cells for each cell line
 Committed stem cells have receptors for specific growth
factors
 Respond to stimulation by division & maturation (precursor
cell stages) into end-stage cells
Precursor cells
-3rd marrow compartment
-Each type of Unipotential stem cell matures into a blast form
[FIGURE 2]
[FIGURE 3]
[FIGURE 4]
[FIGURE 5]
3 MAJOR COMPARTMENTS OR CELL TYPES

1. TOTIPOTENTIAL STEM CELLS
- These cells are present in the first few hours after an ovum
is fertilized. Totipotential stem cells, the most versatile type
of stem cell, can develop into any human cell type, including
development from embryo into fetus.
2. PLURIPOTENTIAL STEM CELLS

- These cells are present several days after fertilization.
Pluripotent stem cells can develop into any cell type, except REGULATORS OF HEMATOPOIESIS
they cannot develop into a fetus.
 Erythropoietin (EPO) - major regulator of erythropoiesis,
3. MULTIPOTENTIAL STEM CELLS stimulates erythroid CFU cells and proerythroblasts
- These cells are derived from pluripotent stem cells. They  Thrombopoietin (TPO) - increases platelet production,
can be found in adults, but they are limited to specific types stimulates megakaryocyte CFU cells
of cells to form tissues. For example, bone marrow stem  Granulocyte CSF (G-CSF) - increases production of
cells can produce all types of blood cells, bone cartilage, and neutrophils, stimulates granulocyte-macrophage CFU cells
adipose (fat) cells.  Granulocyte-macrophage CSF (GM-CSF) - increases
macrophage production, stimulates granulocyte-
STEM CELLS macrophage CFU cells
Stem cells  Interleukins - stimulate B- and T-cell formation, function
- Retain the ability to differentiate into any cell line together with G-CSF and GM-CSF
1. CFU-S [FIGURE 6]
Multipotential stem cells → non-lymphocytes
2. CFU-L
• Secondary multipotential stem cells → lymphocytes
Progenitor cells
-Differentiate into only one cell line
•BFU-E
-CFU-E
•CFU-MEG
•CFU-GM
HEMATOLOGY 1
FIGURE 1
HEMATOLOGY 1
FIGURE 2
HEMATOLOGY 1
FIGURE 3
FIGURE 4
HEMATOLOGY 1
FIGURE 5
HEMATOLOGY 1
FIGURE 6
HEMATOLOGY 1
▪ Cytoplasm: blue-gray to pink gray (production of

LINEAGE SPECIFIC
hemoglobin)
HEMATOPOIESIS ▪ Nucleus
− Round, eccentric
REGULATORS OF HEMATOPOIESIS − Smaller
▪ Erythropoietin (EPO) - major regulator of erythropoiesis, − More condensed
stimulates erythroid CFU cells and proerythroblasts − Stains deeper blue-purple
▪ Thrombopoietin (TPO) - increases platelet production, ▪ 13-30% of nucleated cells in bone marrow
stimulates megakaryocyte CFU cells ▪ This is the last cell division during maturation
▪ Granulocyte CSF (G-CSF) - increases production of
neutrophils, stimulates granulocyte-macrophage CFU cells 4. Orthochromic Normoblast/ (Metarubricyte)/
▪ Granulocyte-macrophage CSF (GM-CSF) - increases (Orthochromic Erythroblast)
macrophage production, stimulates granulocyte-macrophage ▪ Size: 8-12 um in diameter
CFU cells ▪ Cytoplasm: pinker, increased amount of Hb
▪ Interleukins - stimulate B- and T-cell formation, function ▪ Nucleus
together with G-CSF and GM-CSF − Pyknotic
− Eccentric
ERYTHROPOIESIS ▪ 1-4% of nucleated cell in bone marrow
▪ Nucleus is extruded at this stage
• A hemocytoblast is transformed into a proerythroblast
• Proerythroblasts develop into early erythroblasts 5. Reticulocyte
• The developmental pathway consists of three phases Diffusely Basophilic Erythrocyte
st
1 – Ribosome synthesis in early erythroblasts Polychromatophilic Erythrocyte
nd
2 –Hemoglobin accumulation in late erythroblasts and ▪ Size: 7-10 mm in diameter
normoblasts ▪ Cytoplasm:
rd
3 – Ejection of the nucleus from normoblasts and − Pink to pinkish gray
formation of reticulocytes − Still contains small amounts of RNA (polychromasia)
• Reticulocytes then become mature erythrocytes ▪ Nucleus: none
▪ Within 24-48 hrs, the cell loses the organelles &
ERYTHROCYTE NORMAL MATURATION SERIES assumes a biconcave shape
1. Pronormoblast/(Rubriblast)/ (Proerythroblast)
▪ Size: 12-25 um in diameter 6. Erythrocyte
▪ Cytoplasm: ▪ Size: approximately 7.2 um in diameter
− Deeply basophilic ▪ Cytoplasm: Pink
− Relatively small amount ▪ The red blood cell is nonnucleated, round and biconcave
− Perinuclear halo
▪ Nucleus RUBRI NORMOBLAST ERYTHROBLAST
− Large Rubriblast Pronormoblast Proerythroblast
− Round to slightly oval Prorubricyte Basophilic Basophilic erythroblast
− Reddish purple
normoblast
− 1-3 nucleoli
Rubricyte Polychromato- Polychromatophilic
▪ N/C ratio: 8:1
philic erythroblast
▪ 1% of nucleated cells in bone marrow
normoblast
2. Basophilic Normoblast/(Prorubricyte)/ (Basophilic
Metarubricyte Orthochromic Ortochromic
Erythroblast)
normoblast erythroblast
▪ Size: 12-17 um in diameter
Reticulocyte Reticulocyte Reticulocyte
▪ Cytoplasm: intensely basophilic
▪ Nucleus: Mature Mature Mature erythrocyte
− Relatively large Eryhtrocyte Erythrocyte
− Round to slightly oval
− Coarser chromatin LEUKOPOIESIS
− Indistinct nucleoli LEUKOPOIESIS: PRODUCTION OF LEUKOCYTES
− Occupies 75% of the cell
• Leukopoiesis is stimulated by interleukins and
▪ 1-3% of nucleated cells of bone marrow
colonystimulating factors (CSFs)
−Interleukins are numbered (e.g., IL-1, IL-2), whereas CSFs
3. Polychromatophilic Normoblast/ (Rubricyte)/
are named for the WBCs they stimulate (e.g., granulocyte-
(Polychromatophilic Erythroblast)
CSF stimulates granulocytes)
▪ Size: 12-15 um in diameter
HEMATOLOGY 1
• Macrophages and T cells are the most important sources of LEUKOCYTE NORMAL MATURATION SERIES
cytokines 1. MYELOBLAST
• Many hematopoietic hormones are used clinically to • 15-20 um in diameter
stimulate bone marrow • Basophilic cytoplasm
• Round nucleus
FORMATION OF LEUKOCYTES • NC ratio of 4:1
• All leukocytes originate from hemocytoblasts • Fine chromatin
• Hemocytoblasts differentiate into myeloid stem cells and • 2-5 nucleoli
lymphoid stem cells • First recognizable stage in the bone marrow
• Myeloid stem cells become eosinophilic, basophilic and • Makes up 0 to 3 % of nucleated cells in the bone marrow
neutrophilic myeloblasts or monoblasts • MYELOBLAST CANNOT BE DISTINGUISHED FROM
• Lymphoid stem cells become lymphoblasts MONOBLAST IN LIGHT MICROSCOPY
• The myeloblasts develop into eosinophils, neutrophils, and
basophils Type I -N:C ratio of 8:1 to 4:1
• Monoblasts develop into monocytes myeloblast -No visible granules , fine nuclear
• Lymphoblasts develop into lymphocytes chromatin, 2-4 nucleoli
-HSCs, CMPs, and GMPs are not
MYELOID LINEAGE distinguishable with the light microscope
and Romanowsky
staining and may resemble type 1
myeloblast and lymphoblast
Type II Shows dispersed primary (azurophilic)
myeloblast granules in cytoplasm
Type Darker chromatin and more purple
IIImyeloblast cytoplasm
Rare in the BM and can be seen in certain
types of AML
2. PROMYELOCYTE
• 15-21 um in diameter
• Basophilic cytoplasm
• Contains primary non-specific granules
▪ Myeloid lineage gives rise to granulocytes, and monocytes
• N:C ratio of 3:1 to 2:1
(and macrophages), which are not granulocytes. Diagram
• 2-3 nucleoli
does not show changes in cell and nuclear morphologies.
• Chromatin pattern slightly coarser
Band stage is not shown above.
• Normal promyelocyte has a paranuclear halo or “hof”
NON-SPECIFIC GRANULES
✓ Myeloperoxidase
✓ Elastase
✓ Arylsulfatase
✓ Lysosomal enzymes: acid hyrdolases, acid
phosphatase, beta-glucoronidase
✓ Defensins
✓ Cathepsins
✓ Charcoat leyden crystal protein – on
eosinophil only
3. MYELOCYTE
• 12-18 um in diameter
• Appearance of secondary or specific granules
• N:C ratio is 1:1
• Generally shows no nucleoli
• Coarse chromatin
• Last stage capable of mitosis
HEMATOLOGY 1
LYMPHOIESIS
• Lymphocytes are derived from committed stem cells that
originate from pluripotent stem cell.
• Early lymphoid cells further differentiates into B & T
lymphocytes.
LYMPHOID LINEAGE
• Lymphoid stem cell gives rise to T-lymphocyte and B-
lymphocyte lineages
• T-cell maturation - thymus
• B-cell maturation - bone marrow
4. METAMYELOCYTE / JUVENILE CELLS • Plasma cells - present in marrow, lymphatic tissue,
• 10-15 um in diameter connective tissue
• Decreased N:C ratio • B and T cells are non-phagocytic. B- and T-cells are
• Indented or Kidney, peanut shaped nucleus morphologically similar, but functionally different. T-cells are
• Chromatin pattern is coarse and clumped either “helper” or “cytotoxic”. Natural killer (NK) cells are
• Predominant cell in the Bone marrow derived from T-cells and attack tumor cells. T- and B cells
• First stage for the Synthesis of tertiary/ gelatinase granules exit blood at sites of high endothelial venules. Thymus and
bone marrow are primary lymphoid organs.
5. BAND CELLS / STAB CELLS
• 9-15 um in diameter LYMPHOCYTIC MATURATION SEQUENCE
• Elongated or Band shaped nucleus ( C or S shaped) / LYMPHOBLAST
Curved or sausage shape ▪ Cell is similar to other blast cells. It is round or oval, very
• Chromatin pattern is coarsed and clumped large, with a large round to oval reddish-purple nucleus.
• Youngest cell in the series that is normally present in the ▪ The nuclear chromatin material is fine and well distributed
Peripheral blood but perhaps more coarse than in myeloblasts.
• Secretorygranules/ secretory vesicles may begin to be ▪ The nucleus contains one or two nucleoli.
formed during this phase. ▪ The cytoplasm is bluish and non granular and forms a thin
rim around the nucleus
PROLYMPHOCYTE
▪ The nucleus is round or oval in shape but smaller than the
lymphoblast.
▪ The nuclear chromatin is coarse and slightly clumped.
▪ Nucleoli or remnants of nucleoli may be present.
▪ There is an abundant amount of light blue cytoplasm
around the nucleus. Also, there may be a few
azurophilicgranules in the cytoplasm.
LYMPHOCYTE
This is the mature cell of the lymphocytic series and the only
cell form found in the peripheral blood
Lymphocytes vary greatly in size and may be classified as
small, medium or large. However, size does not determine
age of these cells.
The cells are easily distorted and often appear in irregular
shapes in stained preparations. The nuclear chromatin is
condensed to form large, discrete almost solid clumps, with
thickening of the nuclear membrane. Nucleoli are absent.
Non specific granules may be observed in the cytoplasm of
these cells.
HEMATOLOGY 1
MONOPOIESIS
• Development of the monocyte.
• Stages in the monocytic development are:
• Monoblast
• Promonocyte
• Monocyte
MONOCYTIC MATURATION SEQUENCE
1. MONOBLAST
• 12-20 um diameter
• Basophilic cytoplasm
• Non-granular
LYMPHOPOIESIS • N:C ratio of 4:1 or 3:1
• Immunologically competent cells • 1-2 nucleoli
▪ Primary lymphoid organs • MONOBLAST CANNOT BE DISTINGUISHED
▪Bone marrow • FROM MYELOBLAST IN LIGHT MICROSCOPY
▪ Thymus
▪ Secondary lymphoid organs 2. PROMONOCYTE
▪ Lymph nodes • 14-18 um diameter
▪ Spleen • Blue gray cytoplasm
▪ Lymphoid tissues • N:C ratio of 3:1 to 2:1
▪ Lymphocytes • The first recognizable cell in the marrow
▪ B and T lymphocytes
▪ NK killer cells 3. MONOCYTE
• 14-20 um
PLASMA CELL MATURATION • Blue gray cytoplasm
1. PLASMABLAST • Many fine azurophilic granules
• 18 – 25 um in diameter • Ground glass cytoplasm
• Abundant basophilic cytoplasm • Round, kidney shaped, or horse shoe shaped nucleus,may
• Eccentric nucleus show slight lobulation; it may be folded showing brainlike
• Perinuclear halo may be present convultions
• No nucleoli present
2. PROPLASMACYTE • Largest cell in the peripheral blood
• 15 to 25 um
• Intensely basophilic cytoplasm, usually bluer than the blast WHITE BLOOD CELL
stage ▪ NEUTROPHIL
• Eccentric nucleus - 2-5 lobe nucleus
- Primary or secondary granules
3. PLASMACYTE / PLASMA CELL Pink (azurophilic granules)
• 8 to 20 um Grey-blue granules
• Deeply basophilic cytoplasm ( abundant antibodies found in - Life 10 hours
the cytoplasm) Precursors:
• With a Large, well defined hof (perinuclear halo) next to Myeloblast <4%
nucleus Pro myelocytes
• Chromatin is condensed and coarsed Myelocytes
• Nuclues is eccentric and Exhibits “CART WHEEL”- like Metamyelocytes
pattern Band form (stab form)
HEMATOLOGY 1
MACROPHAGES Chemotaxis - movement directed by homing molecules

• These are tissue monocytes DIAPEDESIS
• Cells that usually have an oval nucleus with a net-like • Integrins and selectins are of significant importance in
(reticulated) chromatin pattern. Their cytoplasm is pale, allowing neutrophils to marginate as well as exit the blood
frequently vacuolated, and often filled with debris of and enter the tissues by a process known as diapedesis.
phagocytized cells or organisms Those neutrophils that do not migrate into the tissues
eventually undergo programmed cell death or apoptosis and
NOMENCLATURE OF DIFFERENT MACROPHAGES ON are removed by macrophages in the spleen, bone marrow,
ITS SPECIFIED TISSUE COLLECTION and liver.
• EOSINOPHIL
- Coarser & more deeply red staining granules
- Rarely more than two lobes of nucleus THROMBOPOIESIS: GENESIS OF PLATELETS
- Special role in allergy, inflammation & parasite infection
• BASOPHIL
- Occasionally seen
- Dark cytoplasmic granules
- Role in hypersensitivity response
- Give rise to mast cells
• MONOCYTE
- Larger than lymphocyte
- Oval or indented nucleus
- Monocytes >>>>to macrophage
- Specific function depends on the tissue type STAGES IN THROMBOCYTE DEVELOPMENT
- Kupffer, Oestoclast, Microglia, Langerhans etc. MEGAKARYOBLAST
▪ The cell is large, irregularly shaped with a single or several
• LYMPHOCYTE round or oval nuclei and with a blue, non granular cytoplasm.
- Nucleus is dense, round, oval or slightly indented Nucleoli are usually present.
- B lymphocyte - humoral immunity (~20-30%)
- T lymphocyte - cell-mediated immunity (~60-80%) PROMEGAKARYOCYTE
- Natural killer (NK) cell - cell-mediated immunity (~5-10%) ▪ This cell differs from the megakaryoblast in that there are
- Agranulocyte - lysosomal acid hydrolases bluish granules in the cytoplasm adjacent to the nucleus. The
nucleus in this second stage of maturation has usually
TYPES OF LEUKOCYTES divided one or more times and the cell has increased in size.
GRANULOCYTES
• Neutrophils- 40-70% MEGAKARYOCYTE
• Eosinophils- 1-4% ▪The cell is very large with relatively large amounts of
• Basophils- <1% cytoplasm, and multiple nuclei. The cytoplasm contains
numerous small, uniformly distributed granules that are
AGRANULOCYTES reddish-blue in color.
• Monocytes- 4-8%
• Lymphocytes- 20-45%
Distribution - blood and CT (as transient or wandering cells)

Function - immune protection, movement (cell motility)
Diapedesis - movement out of blood into connective tissue
HEMATOLOGY 1
SUMMARY OF GENERAL MATURATION

CHARACTERISTICS
STAGES IN THROMBOCYTE DEVELOPMENT

▪ Megakaryocytes differentiate from myeloid stem cell and
are responsible for production of platelets.
RED BLOOD CELL
THREE STAGES OF MATURATION OF
MEGAKARYOCYTES RED BLOOD CELL FUNCTIONS
1. Basophilic stage, megakaryocyte is small, has diploid • The main function of the RBC is to carry oxygen to the cells
of the body. Oxygen is transported in a chemical combination
nucleus and abundant basophilic cytoplasm.
2. .Granular stage, here the nucleus is more polypoid, with hemoglobin (HB). The concentration of HB in the blood
is a measure of its capacity to carry oxygen, on which all
cytoplasm is more eosinophilic and granular.
3. Mature stage, megakaryocyte is very large, with approx cells are absolutely dependent for energy and therefore life.
In the tissues, oxygen is exchanged for carbon dioxide,
16-32 nuclei, abundance of granular cytoplasm. It undergoes
which is carried to the lungs for excretion in exchange for
shedding to form platelets.
oxygen.
PLATELETS/THROMBOCYTES • To combine with and transport oxygen, the hb molecule
must have a certain combination of heme (which contains
▪ Small fragments of megakaryocytes
iron) and globin. Deficiencies in the presence or metabolism
▪ Formation is regulated by thrombopoietin
of these substances will result in a decrease in hb- and
▪ Blue-staining outer region, purple granules
oxygen-carrying capacity.
▪ Granules contain serotonin, Ca2+, enzymes, ADP, and
platelet-derived growth factor (PDGF)
RED BLOOD CELL STRUCTURE
NORMAL CELLULAR MATURATION ▪ The biconcave disc shape of a mature rbc is crucial to its
1. CYTOPLASMIC MATURATION function, allowing for close to maximum surface- to –volume
ratio and optimal gaseous exchange. the ability of the
- Immature cell: intensely basophilic (blue) because of high
circulating erythrocyte to perform function of gaseous
RNA content. Loss of blue as cell matures.
st exchange relies almost exclusively on the integral and
- Granules may appear as cell matures. 1 granules are
structural composition and functional capabilities of the
nonspecific. Take on specific characteristics as they mature.
membrane components and sufficient generated atp.
- Relative amount of cytoplasm increases as cell matures.
▪ RBCs are biconcave and average 90fl in volume, with an
2. NUCLEAR MATURATION average surface area of 140 um.
▪ A normal RBC = increase deformability
- Immature nucleus is round or oval and large in proportion to
▪ The RBC membrane is impermeable tocations n, k, and ca.
rest of cell.
it is permeable to water and the anions bicarbonate (hco3)
- As cell matures, decreases in size and takes on varying
and chloride (cl), which freely exchange between plasma and
shapes.
rbc cytoplasm.
- Nuclear chromatin changes from delicate and fine to coarse
and clumped.
THE NORMAL RED BLOOD CELL
- Staining properties change from reddish purple to bluish
purple. • Biconcave disc shape
- Nucleoli gradually disappear. • Aalmon pink
• 7.2 um (7 – 8 um)
3. CELL SIZE • Has an increased life span due to enzyme
- As cell matures, becomes smaller in size. • A nucleated
• Is not rigid and has no inclusions.
HEMATOLOGY 1
MAIN PHYSIOLOGIC FUNCTIONS OF THE RBC − insufficient hemoglobin (e.g., iron deficiency)
MEMBRANE: − reduced availability of o2 (e.g., high altitudes)
• Maintain cell and deformability
• Maintain osmotic balance between plasma and the cell EFFECTS OF EPO
cytoplasm − more rapid maturation of committed bone marrow cells
• Act as supporting skeletal system for surface antigens and − increased circulating reticulocyte count in 1–2 days
receptors • testosterone also enhances EPO production, resulting in
• Aid in the transportation of essential cellular ions and higher RBC counts in males.
gases.
ERYTHROCYTE MEMBRANE
RED BLOOD CELL METABOLISM FUNCTIONS:
• The mature erythrocyte has no nucleus or other organelles • maintain cell shape and deformability
but is capable of existing in the blood circulation for an • maintain osmotic balance bet. plasma and the cell
average of 120 days. An erythrocyte is more limited in cytoplasm
metabolic activity than are other body cells. The cell has a • act as a supporting skeletal system for surface antigens
limited ability to metabolize fatty acids and amino acids and and receptors
lacks mitochondria for oxidative metabolism. • aid in the transportation of essential cellular ions and gases
Composition:
THE LIFE OF A RED BLOOD CELL Proteins -50%
A. Kidneys respond to a lower than normal oxygen Lipids - 40%
concentration in the blood by releasing the hormone Carbohydrates - 10%
erythropoietin.
B. Erythropoietin travels to the red bone marrow and COMPOSITION OF THE ERYTHROCYTE MEMBRANE
stimulates an increase in the production of red blood cells CARBOHYDRATES
(RBCs). • they occur only on the external surface of the red cell.
C. The red bone marrow manufactures RBCs from stem cells • they occur as glycoprotein and glycolipids.
that live inside the marrow. • the antigens of the abo blood group are examples of
D. RBCs squeeze through blood vessel membranes to enter membrane carbohydrates.
the circulation.
E. The heart and lungs work to supply continuous movement LIPIDS
and oxygenation of RBCs. lipid components of the red cell membrane are:
F. Damaged or old RBCs are destroyed primarily by the - 30% free unesterified cholesterol
spleen. - 10% glycerides and free fatty acids
- 60% phospholipids
NUTRIONAL REQUIREMENTS
- Proteins, Lipids, and Carbohydrates PHOSPHOLIPIDS
- Iron, Vitamin B12, and Folic Acid • phospholipids are fat derivatives in which one fatty acid has
- The body stores iron in hemoglobin (65%) been replaced by a phosphate group and one of several
• Intracellular iron is stored in protein-iron complexes such nitrogen-containing molecules.
as ferritin and hemosiderin • phospholipid molecules are characterized by a polar head
• Circulating iron is loosely bound to the transport protein group attached to a non-polar fatty acid tail.
transferrin
CHOLESTEROL
REGULATION OF ERYTHROPOIESIS • the membrane cholesterol is unesterified and lies between
• Too few RBCs leads to tissue hypoxia the two layers of the lipid bilayer.
• Too many RBCs increases blood viscosity • the cholesterol molecule inserts itself in the membrane with
• Balance between RBC production an destruction depends the same orientation as the phospholipid molecules.
on: • the concentration of cholesterol in the membrane is
− hormonal controls animportant determinant of membrane surface area and
− adequate supplies of iron, amino acids, and b vitamins fluidity.
• an increase in membrane cholesterol leads to an increased
HORMONAL CONTROL OF ERYTHROPOIESIS surface area and decreased deformability.
• ERYTHROPOIETIN (EPO) • in extreme circumstances, decreased deformability can
− direct stimulus for erythropoiesis lead to premature rbc destruction.
− released by the kidneys in response to hypoxia
PROTEINS
CAUSES OF HYPOXIA • THESE ARE EITHER
− hemorrhage or increased RBC destruction reduces RBC - PERIPHERAL
numbers - INTEGRAL
HEMATOLOGY 1
1. PERIPHERAL PROTEINS
• the red cell peripheral proteins interact to form a
cytoskeleton.
• the cytoskeleton acts as a tough supporting framework for
the lipid bilayer.
• four peripheral proteins play a key role in the structure of
the red cell cytoskeleton:
- spectrin
- ankyrin
- protein 4.1
- actin
2. INTEGRAL PROTEINS
• these penetrate the lipid bilayer and are firmly anchored
within it.
- band 3
- glycophorins a, b, and c
- na+/k+ atpase
- glucose transport protein
- surface receptors
MEMBRANE ENZYME SYSTEM

1. Na-K ATPASE
-controls transport of sodium and potassium
2. Ca-Mg ATPASE
-calcium pump moves calcium out of the cell to plasma
against a high concentration gradient
RED BLOOD CELL DESTRUCTION

ERYTHROCYTE DESTRUCTION
• as the red cell ages, changes occur that make it susceptible
to destruction.
• loss of sialic acid and lipids, decreased atp and increased
calcium have been implicated in the aging process.
• at 120 days the erythrocytes are recognized as abnormal
and are removed by phagocytic cell in the res.
• as the cell ages it is depleted of glucose and their surface
ENERGY METABOLISM OF ERYTHROCYTE area decreases.
ENERGY REQUIREMENT: • the spleen recognizes abnormalities in the cell and
• the cell requires energy for cell metabolism and to preserve sequester it for removal.
the membrane integrity • culling is a splenic function wherein old/aged/senescent
• various enzymatic reactions in the cell require energy. redblood cells are filtered and destroyed through the
• energy is required to reduce proteins and maintain phagocytosis of splenic macrophages.
hemoglobin in its reduced state for proper functioning.
• two site prone to oxidation are the iron atom in the heme AS AN ERYTHROCYTE AGES, THE FOLLOWING
ring and the sulfhydryl groups on the globin molecule. PROCESSES OCCUR:
• oxidation of the normal ferrous state to the ferric state 1. The membrane becomes less flexible.
results in methemoglobin which does not deliver oxygen 2. The concentration of cellular hemoglobin increases.
• normally 1% to 3% of oxygen is oxidized to methemoglobin 3. Enzyme activity, particularly glycolysis, diminishes.
• oxidation of sulfhydryl groups causes hemoglobin
precipitation (heinz body formation)
HEMATOLOGY 1
TYPES OF RBC DESTRUCTION

A. EXTRAVASCULAR HEMOLYSIS
▪ 90% of RBC destruction
▪ Also known as the macrophage mediated hemolysis
▪ As old or damaged RBCs are phagocytized, hemoglobin
molecules are disassembled.
▪ IRON – returned or erythroid precursors in the bone
marrow or stored as ferritin or hemosiderin within the
macrophages
▪ GLOBIN – broken down and recycled into amino acid pool
▪ PROTOPORPHYRIN – disassembled with its alpha carbon
exhaled in the form of carbon monoxide while biliverdin is
converted to bilirubin and eventually urobilinogen which is
then excreted in the feces and urine.
▪ Happens when complement is not activated or incompletely
activated.
▪ Associated with Rh hemolysis
▪ INCREASED UNCONJUGATED BILIRUBIN
▪ INCREASED URINE AND FECAL UROBILINOGEN
▪ PBS: SPHEROCYTES
B. INTRAVASCULAR/FRAGMENTATION HEMOLYSIS
▪ ≤10% OF RBC DESTRUCTION
▪ Occurs within blood vessels
▪ RBC ruptures releasing hemoglobin directly into the
bloodstream
▪ Hemoglobin disssociates into alpha-beta dimers picked up
by the protein carrier haptoglobin and brought to the liver for
processing (same process as extravascular).
▪ When haptoglobin is depleted, unbound hemoglobin
appears in the plasma.
▪ Free hemoglobin also appears in the urine
▪ Hemoglobin not excreted by the kidney or bound to
haptoglobin is oxidized to methemoglobin.
▪ Happens when complement is completely activated
▪ Associated with ABO hemolysis
▪ Decreased haptoglobin and hemopexin
▪ Hemoglobinemia
▪ Hemoglobinuria

Hema Lec Trans Prelim

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Hema Lec Trans Prelim

Uploaded by

Copyright:

Available Formats

OUR LADY OF FATIMA UNIVERSITY

MEDICAL LABORATORY SCIENCE BATCH 2024

COURSE OUTLINE: PRELIMS

- primarily deals with the cellular elements of the peripheral

Hematology - study of the clinical, morphologic and

BLOOD - red liquid that circulates in the arteries and veins of

Total blood volume in an adult is 5 to 6 liters or 7 to 8% of

The clinical hematology section perform scientific

II. DEFENSIVE FUNCTIONS HANDWASHING

BASIC HEMATOLOGICAL QUALITY ASSURANCE PROGRAM

Reticulocyte count Sites of skin puncture

Methods of Skin Puncture

Sites to avoid ETS METHOD (EVACUATED TUBE SYSTEM)

Reasons for capillary puncture in infants and very young

3 factors involved in a good venipuncture

Key notes to remember on Venipuncture

Sites of venipuncture 3 y/o to adult life

IN OLDER CHILDREN (18 MONTHS TO 3 Y/O)

VEINS OF THE ANTECUBITAL FOSSA

ORDER OF DRAW FOR VENIPUNCTURE

Advantages of Evacuated Tube Method

MOST COMMON ANTICOAGULANT USED IN 3 forms of oxalate:

DISADVANTAGES: Not recommended for peripheral blood smear

STAGES OF HEMATOPOIESIS ADULT HEMOGLOBIN GLOBIN CHAIN

Significant Contributions of Other Organs:

1. RED MARROW, which is hematopoietically active marrow,

MAJOR SITE OF HAEMATOPOIESIS

STRUCTURE AND DEVELOPMENT

INDICATIONS FOR BONE MARROW STUDIES

BONE MARROW EXAMINATION

MYELOID ERYTHROID (M:E RATIO)

Scenario M:E ratio Interpretation

NORMAL MARROW CELLS

STEM CELL THEORY OF HEMATOPOIESIS

STEM CELL THEORY OF HEMATOPOIESIS

3 MAJOR COMPARTMENTS OR CELL TYPES

2. PLURIPOTENTIAL STEM CELLS

▪ Cytoplasm: blue-gray to pink gray (production of

MONOCYTIC MATURATION SEQUENCE

MACROPHAGES Chemotaxis - movement directed by homing molecules

Distribution - blood and CT (as transient or wandering cells)

SUMMARY OF GENERAL MATURATION

STAGES IN THROMBOCYTE DEVELOPMENT

MEMBRANE ENZYME SYSTEM

RED BLOOD CELL DESTRUCTION

TYPES OF RBC DESTRUCTION

You might also like