HEMATOLOGY 1 Reverse Isolation – isolates a person who is

immunocompromised or has a low immune

system to avoid being
infected by a disease

Ex. Elderly, Pregnant women, children and etc.

● Anton van Leeuwenhoek

ECCENTRICITY - Father of Microbiology
- First to discover and described Giardia
It is the measure of how much the cell lamblia
deviates from being circular Old man wearing
glasses Falling leaf
HISTORY OF CLINICAL movement
HEMATOLOGY

● Athanasius Kircher
- In 1646, he uses a microscope to ● Giulio Bizzozero - Discovers:
observe plague victims H. pylori
- In 1658, he observed the bacteria Functions of
Scrutinium Pestis (also known today as platelets
Yersinia pestis) which he described as
“little worms” or “animalcules” in the
blood
● James Homer Wright
- Proposed hygienic
- He is a pathologist who works at a
measures
Pathology Laboratory at Massachusetts
Isolation
General
Quarantine
Hospital
Burning clothes are
- In 1902, he modified
worn
the
by the infected
Rowanowsky stain by using the
Wearing facemasks
Wright stain
Difference between Isolation and - Megakaryocytes are where the platelets
Reverse Isolation origin

Isolation – isolates a person who is

infectious or infected by a disease Ex.
● Paul Ehrlich
Tuberculosis patient - A German Bacteriologist
- He classified leukocytes
- Invented the cure for Syphilis –
Salvarsan in 1910
 ● William Hewson Ø In vitro (inside a tube)
- A British Anatomist and
Physiologist - Red in color – due to the presence of
hemoglobin
- Described Blood Coagulation
- Thick and viscous – because it has
- Isolated Fibrinogen
many compounds
- Father of Hematology
- Slightly alkaline – pH 7.35 –
7.45
- Specific gravity – 1.045 – 1.065
HEMATOLOGY
- 7 – 8% of the total body weight
Comes from the word HAIMA which means blood - Total volume: [ M = 5 – 6L (a lot of blood
and LOGOS which means study because of the body structure); F = 4 –
5L (lesser because the presence of
Blood menstruation)]

- It is a nutritive fluid Composition of Blood

- It contains a lot of components
- Liquid (Plasma non-clotted blood/Serum
● glucose
clotted blood)
● proteins
- Water (91.5%)
● vitamins
- Chemicals
● minerals
- Proteins (7%)
- Participates in the physiologic and
- Others (1.5%)
pathologic activities of the body
● Electrolytes
Arterial blood – bright red w/oxygen
● NPN (Non-protein
Venous blood – dark red w/o oxygen Nitrogen
Components)
Functions of Blood ● Hormones and
Enzymes
● Respiration/Transport
● Food Materials
● Carbon Dioxide (cells to lungs) ●
Oxygen (lungs to tissues/cells)
● Hormones and Nutrients
● Nutrition - Solid/Cellular
● Excretion components
● Homeostasis ● RBC
● Body protection - 45% of blood
- WBC – fights diseases volume
- Platelets – blood - 3.8-6 x 1012/L
coagulation (M/mL)
● WBC
Characteristics of Blood - 3.6-10.6 x 109/L
(T/mL)
- Fluid in vivo (inside the body)
 - Granular and Agranular
- Platelets
● 150 – 450 x 109/L (T/mL)
-Granulocytes and Agranulocytes
● Granulocytes
- Basophils [0.5 – 1%] à
Mast Cells (In Tissues) -
Eosinophils [ 2 – 4%]
- Neutrophils [60 – 70%]
● Agranulocytes
- Lymphocytes [20 – 25%]
- T cells (Thymus) à Th
(Helper), Tc (Cytotoxic),
Ts (Suppresion), Tm (Memory), Tdh
(Delayed
Hypersensitivity)
- B cells (Bone Marrow) à plasma
and memory
cells (20 years lifespan)
- Monocytes [3 – 8%] à
Macrophages
Tissues)
10 SERVICES OFFERED BY HEMA
AND HEMOSTASIS LAB
● Specimen collection & preparation for
exam
● Quantitative manual & instrumental
measurements of
cells
● Measurements of cells
● Measurements of cell volumes
● Evaluation of cellular contents &
components
● Cellular identification
● Identification of reactive or neoplastic
alterations of cell populations
● Evaluation of leukocytes, erythrocytes
and platelet function
● Evaluation of cellular development and
formation (BM)
● Evaluation of hemostatic function
(In PREFIXES
● a- / an- (lack, without) (absent,
decreased)
● aniso- (unequal) (dissimilar)
● ante- (before) ● Brady- (slow)
● Cyto- (Cell)
● Dia- (Through)
● dys- (abnormal) (difficult, bad)
● erythro- (Red)
● ferr- (Iron)
● hemo- (pertaining to blood)
● hyper- (above, beyond)
(extreme)
● hypo- (beneath, under)
(deficient, decreased)
● iso- (equal, alike, same)
● leuko- (white)
● macro- (large, long)
● mal- (bad, abnormal)
● mega- (large, giant)
 ● meta- (after, next) (change) ● centesis (surgical puncture to remove
● Mono- (one) fluid)
● morph- (Shape) ● oxia (oxygen level)
● myel(o) (From the BM) (Spinal cord) ● algia (pain)
● pan- (all, overall) (all-inclusive)
● phleb- (vein)
● phago- (eat, ingest) EXAMPLES OF HEMATOLOGIC
● poikilo- (varied, irregular) TERMS
● schis- (split)
● scler- (hard) ● Anisocytosis (An + iso + cyt + osis)
● splen- (Spleen) ● Aplasia (A + plasia)
● Thromb(o)- (Clot, Thrombus) ● Anemia (An + emia)
● xanth- (yellow) ● Dysmyelopoiesis (Dys + myelo
● peri (around) + poiesis)
● cata (down) ● Panmyelosis (Pan + myel(o) + osis)
● epi (on, over, upon)
● anti (against) QUALITY ASSURANCE

Coordinated effort to organize la activities

SUFFIXES
Provide best services to patients and
● -blast (primitive) doctors
● -cyte (cell)
● -ectomy (excision, cut out) Involves control and monitoring

● -emia (blood) ● Staff competence (quarterly

● -itis (inflammation) assessment)
● -lysis (destruction, dissolving) ● Materials and methods (manual
● -(o)logy (study of) updated, area that can be ready
● -oma (swelling, tumor) available) ● Reporting of
results:
● -opathy (disease)
➢ TAT (turnaround time)
● -osis (state, condition, increase)
➢ STAT –Short
● -penia (decrease, lack of)
Turnaround Time (urgent,
● -phil(ic) (attracted to, affinity for) as soon as possible)
● -plasia (-plastic) (cell production or (30% costly)
repair) ● Satisfaction of clients
● -poiesis (cell production, formation, - (RR: receiving and releasing area)
development) ● Financial costs: revenue
● -poietin (stimulated production)
● -stasis (same, standing still)
● -trophy (nourishment)
● spasm (twitch)
 QUALITY CONTROL PRECISION

- Closeness of results obtained from repeated

analysis of same patients
- Reproducibility
- Eg. 199, 200, 201
- Use 10 runs for 10 consecutive days

CONTROL

- Predetermined assay valueà ginapalit

- Same matrix as patient sample (ex. same as
blood)

PRIMARY STANDARD DELTA CHECKS

- Calibrate instrument - Assesses change

- Certified reference material - Compare result from sample
- Essentially pure form analysis with result of previous sample
- Fixed and known composition - Same analyte, same patients
- Ex. Dengue patients for instance
SECONDARY STANDARD
(logbook if unautomated record)
LINEARITY
- Analyte concentration ascertained by
reference to a 10 std
- The ability to generate results proportional
to the calculated conc or activity of the
CALIBRATOR
analyte
- Preserved humn/ surrogate cell suspension - (medtech will dilute 5 sample and runà
- Determined hematologic parameters computeàgraph)
LEVELS OF CONTROL
RELIABILITY
a. Normal—blue color
- Extents to w/c mtd is able to maintain
accuracy and precision over time
b. Normal high—green
- (maintain the laboratory performance)
c. Abnormal high—red
REFERENCE INTERVAL
ACCURACY
- Reference range
- Closeness to the true/ actual value - Range of values of analyte in healthy
- Eg. Exact 200, 100, etc individuals
DIAGNOSTIC SENSITIVITY
- Proportion of patients with the disease with a
positive result
- (Eg. Positive si patients sa dengue dapat
ang result kay positive sa dengue)
DIAGNOSTIC SPECIFICITY
- Proportion of patients identified correctly by
the test as not having the disease
SYSTEM ERRORS
- Errors within test systems
- (Eg. Malfunction of machine, calibrations)
RANDOM ERRORS
- Occur w/o prediction regularity
- (Brownout, segeg tuplok2, wala na tarung ug
shutdown si machine)
TYPES OF ERRORS
A. PREANALYTICAL ERRORè before testing
- Specimen obtained from wrong
patients
- Specimen produced at the wrong
time
- Specimen collected in the wrong
tube
● Purple top—EDTA—
Ethylen
ediamine tetraacetic acid
● Light Blue top—SODIUM
CITRATE
- Blood sample collected in wrong
order
- Incorrect labeling
● label after collection, never
pre label
- Improper processing of specimen
● check always the request
B. ANALYTICAL ERRORè during testing
- Oversight of flaggingà blink blink
- Out of control QC resultsàif dili mo
pasok sa range dili mag run ug
result kay mali na siya
- Wrong assay performed
● dapat mag control bago mag
run
C. POSTANALYTICAL è after
testing
- Verbal reporting resultsà I relay tama ang
message
- LIS (lab info system) incompatibility error
- Confusion about reference rangesà butang tama
ang reference range/equivalent
- Failure to report critical values
● ASAP (call the attending
physician/nurse)
INTERNAL QUALITY CONTROL
- The actual running of control by the staff
- Internal to the laboratory
● run control material, process, and put
the result in the log book
EXTERNAL QUALITY CONTROL
- NEQAS—National External Quality
Assessment Scheme
- NEQAS sent unknown specimen
sample tas I examined if pasado ba si lab mo
operate or dili, need 90% for passing.
- Need for licensed to operate DISCUSSION 3
 BASIC HEMATOLOGICAL TECHNIQUES HEMOGLOBIN

- Estimated by:
OBJECTIVES ● Color
● Power of combining with oxygen or
- Define Complete Blood Count (CBC) and its carbon monoxide
inclusive tests. ● Iron content
- Discuss the advantages and disadvantages - 1 gram Hb = 1.34 mL of oxygen
of manual and automated methods of CBC. - Detection and assessing clinical anemia
- Define Hemoglobin and Hematocrit and - Gold Standard: Hemiglobincyanide
other tests and explain their clinical
(HiCN) or Cyanmethemoglobin Method
significance.
- Lab: Acid Hematin Method = 0.1N
- Elaborate rule of three and its use.
Hydrochloric Acid
- Compute for RBC indices using the formula.
- Manual - Reagent: Drabkin’s Reagent (Pale
Yellow) - Reference Values:
● Adult Male: 14.0 - 18.0 g/dL
COMPLETE BLOOD COUNT (CBC) (140 - 180 g/L)
● Adult Female: 12.0 - 16.0 g/dL (120
- Foundation procedure - 160 g/L)
- Can tell various disorders (hematological or ● Infants: 10.0 -14.0 g/dL (100 - 140
hematologic manifestations secondary to other g/L)
diseases) → abnormalities in RBCs, WBS, ● Newborns: 15.0 - 20.0 g/dL
and/or platelets - Tests: (150 - 200 g/L)
● Hemoglobin (Hb) ● HEMATOCRIT
Hematocrit (Hct)
● RBC count with morphology (Not - Packed Cell Volume (PCV)
done manually due to medtech - Simple screening test for anemia
error [random error]) - Reference method for calibrating blood count
● WBC count with differential systems
● Platelet estimate - Rough guide to accuracy of Hb
● RBC indices measurements
TECHNIQUES IN PERFORMING CBC - Used in the calculation of red cell indices
- Reference Values:
Advantages and Disadvantages: ● Male: 0.47 L/L (+/- 0.07)
○ Normal Values = 40
Manual
(0.47 - 0.07) to 54
- Low Cost (0.47 + 0.07) %
- Labour Intensive ● Female: 0.42 L/L (+/- 0.05)
○ Normal Values = 37 -
Automated 47 %
● SI Unit = L/L
- High Capital Costs ● Common Unit = %
- Rapid Performance - More precise
RULE OF THREE MCHC = (Hb x 100) / Hct = ___g/dL

- 3X Rbc = Hb +/- 0.5 ● Hb conc. or color of the average

- Ex: 3.3 RBC x 3 = 9.9 RBC
- 9.9 - 0.5 = 9.4 ● Reference Values: 32-36 g/dL
- 9.9 + 0.5 = 10.4 ● Hypochromic = Below 32
- 3X Hb = Hct +/- 3% (commonly used) ● Normochromic = Normal (Within
- Ex: 11.0 Hb x 3 = 33 32-36 g/dL)
- 33 - 3 = 30 ● Hyperchromic = Above 36
- 33 + 3 = 36 ● Ex: (9.0 x 100) / 42 = 21 (BELOW
- BUT, only applicable with red blood cells in NORMAL VALUES → Hypochromic) BLOOD CELL
normal size (7-12) and color COUNTS

- Anemia or Polycythemia ●
Reference Values:
RED CELL INDICES ○ Male: 4.6 - 6.0 x
10^12/L
- morphologic classification of anemias
○ Female: 4.0 - 5.4 x
- Used also in QC
10^12/L
- calculated from Hb, Hct, and RBC count
- Leukocytosis or Leukopenia
- 3 indices:
● Indicate Infection, follow
● MCV (Mean Corpuscular
disease progress
(Cell) Volume)
● Reference Values:
● MCH (Mean Corpuscular
0 4.5 - 11.5 x 10^9/L
Hemoglobin)
(Rodak) -
● MCHC (Mean Corpuscular
Thrombocytosis or
Hemoglobin
Thrombocytopenia
Content/Concentration)
● Associated with hemostatic
mechanism
MCV = (Hct x 10) / RBC Count =
___femtoliter ● Reference Values:
0 150-450 x 10^9/L
● Volume or size of average RBC (Rodak)
● Reference Values: 80-100 fL ● below
80 = microcyte (Iron Defeciency Anemia);
above 100 = macrocyte (Megaloblastic
LEUKOCYTE DIFFERENTIAL COUNT
Anemia)
● Microcytic (Small)
● Absolute (x10^9/L) = multiply relative
● Normocytic (Normal) number of WBCs by the total WBC count
● Macrocytic (Big) ● Example:
● Ex: (48 x 10) / 5 = 96 (NORMAL) - ABSOLUTE COUNT

MCH = (Hb x 10) / RBC Count = ___pictogram Total WBC = 5.0 x 10^9/L

● Weight of Hb in average RBC Differentials:

● Reference Values: 27-33 pg
● Correlated to MCV ○ Neutrophils: 75% - (0.75)
 (5.0) = 3.75 ● 0 - 50 y/o (Males) = 0.15mm/hr
○ Lymphocytes: 20% - (0.20) more than 50:
(5.0) = 1.0 0-20 ● 0 - 5 y/o (Females) =
○ Eosinophil: 4% - (0.04) (5.0) = 0.2 0.20mm/hr more than 50: 0-30
○ Monocyte: 1% - (0.01) (5.0) =
0.05
DISCUSSION 4
100 % = 5.0
RETICULOCYTE COUNT

SPECIMEN COLLECTION BY VENIPUNCTURE

● Immature RBCs that lost the nucleus but
retain RNA
● One stage earlier than the mature
PHLEBOTOMY VENIPUNCTURE
RBC
● Juvenile red cells demonstrated by - Most common technique used to obtain
Supravital stains blood.
● Reflects erythropoietic activity of BM - Requires ample skill to ensure accurate
● Manual - Stains: Brilliant Cresyl Blue results and preservation of patient vein
(usual); Methylene Blue integrity
● Relative count (percentage) = - Incision in the vein of the arms
○ Adult: 0.5 - 1.5 % - Three Methods: Syringe, Buttefly, ETS
○ Neonates: 1.5 - 6.5 % (Evacuated Tube System)
● Absolute Count (x 10^9/L) = 50 x
10^9/L
INITIAL STEPS

ERYTHROCYTE SEDIMENTATION RATE - Correct Patient Identification (1st Critical Step)

● Compare info in the lab request
- Measure of degree of settling of RBC in ● Verify the age and sex
plasma in an anticoagulated whole blood spx ● Wrist Band (Admitted)
during specified period of time. ● IV Bottle - Note Isolation
- test for the non specific inflammation Restrictions - Note dietary
- run in an hour - Two Methods: restrictions (e.g.
● Westerngren Fasting)
● Wintrobe mtd - Reassure the patient, don’t deceive
● Nowadays: Modified Westergren (Do not tell that it does not hurt)
- Pipette, Vial, Rack (Modified - Position the patient
Westergren) ● Lie in bed for admitted px
● For ambulatory px in an extraction chair

- Normal Values - Westergren

- Assemble supplies accessibility - SPECIMEN TRANSPORT
- Selection of the site ● Proper handling and timely
transport
● Median Cubital (Preferred)
- Proper way of removing gloves (Turgeon
● Median Basilic
page 8, 5th Edition)
● Cephalic veins
● If not veins of the wrist, back of the
hand, ankle or foot (Consult if there is
circulatory problems) -Points:
● Avoid areas with hematoma, burns
(sensitive, easy portal of entry to
pathogens to create infection), scars or
edema, IV line, mastectomy site also
- Apply the tourniquet 3-4 inches above the site
- Clean the site properly (apply firm but gentle
pressure —> start from center and work COMMON DURING VENIPUNCTURE PROCEDURE
downward and outwards covering an area of
2cm or more) (Blood Culture: Povidone Iodine; 1. Failure to dry site
do not use in others for it will falsely increase 2. Bevel down
the potassium, phosphorus, and uric acid) 3. Failure to mix blood ASAP
● READ WHO PHLEBOTOMY 4. Failure to release tourniquet; and
GUIDELINES 5. Excessive pulling of plunger
- Inspect the needle and the vacuum
- Release the tourniquet
● Always before removal of needle
- Remove needle and Apply pressure SPECIAL SITUATION ENCOUNTERED
● Do not allow px to bend the arm,
- IV Site/Med Lock
this will reopen the wound and
● Never drawn on the same side
result to haemorrhage
● Opposite Arm or Distal or below IV
- Discard needle (in Puncture Proof Container /
line only
Sharp Container)
● Both arms with IV lines: Stop for at
- LABEL SPECIMEN
least 2 or 5 mins; Note in the
● No pre-labeling
request form also; Tell the nurse in
● Name, Date and Time, duty
Age/Sex, Initials of phleb, and any - Transfusion
info required by the institution
 ● May collect blood SOURCES OF ERROR
● Should be indicated and drawn
opposite arm - Hemolysis
● Note in the request form - HD - Failure to dry the site
Shunt/Fistula - Failure to wipe the 1st drop of blood
● Avoided - Vigorous massaging or milking the area
● Ask for the status may draw distal - Accidental capturing of bubbles
atleast 4 inches below
- Mastectomy
● Collected on the other side
● Picture of the proper site of skin puncture
● Mastectomy promotes lymphostatis
(Turgeon)
- Hematoma, Burned, or Scarred
Areas
● Avoided
● Scarring causes veins difficult to
palpate

SPECIMEN COLLECTION BY SKIN

PUNCTURE

● Burned areas are prone to infection

- REMEMBER: Each phlebotomist is allowed
only two attempts. THEN, what to do next?
Call for the other assigned Medtech in the
lab, ideally,
Senior Medtech
- Done in infants particularly newborns
- Hospital induced anemia, osteomyelitis -
puncture not more than 2.4mm
- Safer
- Puncture site should be warm
- Sites: 3rd or 4th finger, big toes, heel, and
last resort is the earlobe
- For adults because of obesity, burns,
extremely small veins
- Blood is a mixture of capillary, venous and
arterial blood with interstitial and intercellular
fluid
- It is important to wipe the first drop of blood
(Because it has tissue juices)
- A good puncture requires no forcing or hard
squeezing
DISCUSSION 5 fetus. Versatile except for that
becoming a
HEMATOPOIESIS fetus)
3. Multipotential (derived from
(Periods / Phases of Hematopoiesis) pluripotent cells. Found in adults
and are limited to specific types of
(Adult Hematopoietic Tissue)
cells to form tissues. Not so
(Stem Cell Theories and Cell Production) versatile for it is limited to what type
of
cells)
HEMATOPOIESIS
HEMATOPOIETIC DEVELOPMENTAL
- the process of blood cell production, PERIODS
differentiation, and development - restricted in the
bone marrow 1. Mesoblastic Period/Phase
- A.K.A. yolk sac
hematopoiesis
- 19-20 days of gestation (2nd week)
ORIGIN OF BLOOD CELLS - Occurs in blood islands of the yolk
sac
- Hematopoietic Stem Cells (HSCs) - Primitive erythroblasts (Found in
● Foundation of adult yolk sac arising from mesodermal
hematopoietic system
cells.
● Capable of repopulating
Occurs intravascularly [the process
(repopulation)
is inside the blood vessel].
● Now widely accepted that it was Angioblasts [forms future Blood
produced by the embryo
Vessels])
- Types of Human Stem Cells
- Forms three types of hemoglobin,
1. Totipotential (present in the 1st few namely,
hours after an ovum is fertilized.
Portland (2ζ 2γ),Gower I (2ζ
The most
2ε), and Gower II (2α 2ε)
versatile)
2. Hepatic Period/Phase
2. Pluripotential (present several days
- Characterized by low levels of
after fertilization. Can develop into hemoglobin A
any cell type except into being a - 4th to 5th (Rodak) or 5th and
6th week of gestation
 - Characterized by recognizable (hematopoietically inactive
clusters of erythroblasts, marrow)
granulocytes, and monocytes - Yellow Marrow = inactive ADULT
- Definitive morphologic
hematopoiesis - presence of HEMATOPOIETIC TISSUE
erythroblasts + lymphoid
- Organs involved: Bone Marrow, Lymph Nodes,
cells
Spleen, Liver, Thymus
- Fetal liver: Primary erythroid organ
- Primary Hematopoietic Tissue: Bone
(extravascular outside the blood
Marrow, Lymphoid
vessel);
- Primary Lyphoid Tissue: Bone Marrow
Retaining the activity from
(B-Cells)and Thymus
1st to 2nd week after birth (T-Cells)
- Spleen: Involved solely in - Secondary Lymphoid Tissue: Spleen, Lymph
lymphopoiesis Nodes, GALT
- Starting of megakaryocyte (Gut-associated Lymphoid Tissue)
production
● Bone Marrow
- Fetal Hgb = 2 alpha and 2 gamma
- Tissue within cavities of cortical
- Hb A1 and Hb A^2 are also
bones
detected (low)
- 2 Types:
- Other organs that will be activated:
- Red Marrow
Spleen, Thymus, Lymph Nodes,
(Hematopoietically
(Kidney)
Active; Flat Bones & Proximal
- Fetal hepatic hematopoiesis = Adult
ends of long bones: Sternum
extramedullary hematopoiesis
(breast bone), Skull, Ribs, Pelvis,
(outside the bond marrow): Rare,
Pelvic Bones, Scapula in the
and
clavicle, Femur);
occur if there is BM failure - Yellow Marrow
3. Myeloid/Medullary Period/Phase (Hematopoietically
- 5th month of gestation Inactive; Adipocytes;
- Developing bone marrow cavity - Normally(Adult) 50% RM, 50%
(long bones): Inner part of BM YM)
- During the end 6th month, the BM - (Rodak) Between 5 to 7 years old,
will become the primary site of yellow marrow will replaced red
hematopoiesis marrow =
- Measurable amounts of the Retrogression
following: EPO ● Liver
(Erythropoietin), G-CSF - Major site in hepatic
(Granulocyte-colony hematopoiesis (fetus)
stimulating factor), - Hematopoietic functions
GranMacro-CSF (adults):
(Granulocyte-Macrophage - Synthesis of various transport CHON
Colony-Stimulating Factor), HbF, - Storage of Minerals and Vitamins
HbA2 and Hb A1 - Bilirubin Conjugation - Bilirubin
- 4th year of life = marrow replaced by Transportation - Pathophysiology:
fats - Porphyrias (Enzymatic
 deficiencies; removed, platelets will increase as you got
Accumulation of examine with
intermediary porphyrins) CBC for the platelets will just go on and forth
- Increase bilirubin since spleen was removed.
conjugation, Fe++ storgae - Pathophysiology:
(Severe H.A. - Hemolysis
and RBC dysplasias) (Increased environmental stress due to
- Extramedullary small
hematopoiesis (Bone openings by
marrow failure) inter endothelial junction)
- Storage Diseases - Splenomegaly
(Monocytes/macroph ages (Enlarged and
[Kupffer Cells, Enzymatic Palpable; Conditions:
deficiencies, Chronic
Hepatomegaly with Leukemia, Genetically defective
liver dysfunction) RBCs, Malaria)
● Spleen - Hypersplenis m (Enlarged leading
- Largest lymphoid organ to
- Vital but not essential for Pancytopenia caused commonly
life by
(mabuhi ka if wala siya) - congestive splenomegaly secondary
Functions: to liver cirrhosis and portal
- Indiscriminate filter of hypertension)
blood ● Lypmh Nodes
- Note: In a healthy - Bean shaped member of the
individual, the spleen lymphatic system occurring in
contains about groups or in chains and are
350mL of blood located superficially or deep
- Sequesters senescent - Functions:
(old) red cells: Culling - Formation of new
(Cells are phagocytozed lymphocytes
with subsequent - Processing of specific
immunoglobulins
degradation of cell
- Filter particulate matter, debris,
organelles), Pitting
bacteria entering via lymph
(Splenic
- Pathophysiology:
macrophages will remove
- Adenitis (Infection of lymph
the damage surface
node)
membrane from the
- Infiltration by malignant
circulating RBC)
cells =
- IgM synthesis
metastasis
- Storage for platelets (⅓)
● Thymus
- Note: In healthy individual,
- Efficient, well-developed at
approximately 30% of the total platelet count is
birth
sequestered in the spleen; so if the spleen will be
- Pathophysiology:
- Non-developmet
 (gestation) - Lack of T-
lymphocytes,
uncontrollable infections,
death
- Thymic disturbance
(adults) - No effect
STEM CELL THEORIES
● Pluripotential Stem Cell
- One cell gives rise the varied blood
cells (Eryhtroid,
Lymphoid, Myeloid )
- Widely accepted theory
● Commited Stem Cells
- Give rise to descendants or
progeny cells that eventually become
restricted to
specific cell line development
CHARACTERISTICS / FATES OF STEM CELLS
- Capable of self renewal
- Give rise to differentiated progeny
- Able to reconstitute the hematopoietic
system of an
irradiated host
- Apoptosis
CYTOKINES (HEMATOPOIETIC GROWTH
FACTORS
- Group of specific glycoCHON
- Regulates the proliferation, differentiation,
and maturation of hematopoietic precursor
cells
- Include interleukins, lymphokines,
monokines, chemokines, interferon, and
colony stimulating factors
COLONY STIMULATING FACTORS
(CSFs)
- Produced by many cells
- High specificity to target cells
- Active at low concentration
a - Example: G-CSF: Primary target is
granulocyte
- Can also have synergistic effect (IL-3 and G-
CSF for Megakaryocyte colony stimulation)
INTERLEUKINS
- Numbered in order of identification -
Characteristics:
- CHON that exhibit multiple biologic activities
- Synergistic interactions with other cytokines
and growth
factors
- Interacting systems with amplification
potential
- Effective at very low concentrations
- Refer to Page 72-73 of Hema Book 3rd Ed
by RODAK for lists of interleukin with
sources, target site and their roles
FACTORS THAT STIMULATE LINEAGE SPECIFIC
HEMATOPOIESIS
● Erythropoiesis
- Erythropoetin
- Hypoxia
● Leukopoiesis
- Colony stimulating factors
- Interleukins-(IL3)
● Megakaryocytopoiesis
 - Thrombopoietin - Stimulating hormonal factor is TPO which is
- Meg-CSF mainly produced by the liver - EPO = kidney,
ERYTHROPOIESIS TPO = liver

- Occurs in BM
- CFU-GEMM (Colony Forming Unit
DISCUSSION 6
Granulocyte, Erythroid Cell, Monocyte, and
Megakaryocyte) gives rise to earliest
identifiable colony of RBC = Burst Forming NORMOBLASTIC MATURATION
Unit
- Erythroid (BFU-E)
RED BLOOD CELLS
- BFU-E will become CFU-E that has many
EPO receptors
- called erythrocytes
- EPO induces Hb synthesis
- its 1 true function: caryy oxygen from lungs
to tissue
LEUKOPOIESIS
- 3 nomenclature
- Major Categories: - Erythroblast terminology
(Europe)
1. Myelopoiesis
- Normoblastic terminology
- GM-CSF
(USA)
(Granulocyte-macrop hage
- Rubriblast terminology (some since it
colony-stimulating
parallels the WBC development
factor), G-CSf
nomenclature
(Granulocyte colony
stimulating factoR), M- NOMENCLATURE FOR ERYTHROID
CSF (Macrophage PRECURSORS
Colony-Stimulating
Factor) including IL-3, IL- Normoblastic
5, IL-11
- IL-3 multineage: - Pronormoblast
stimulating granulo, mono, - Basophilic Normoblast
mega, and - Polychromatic (Polychromatophilic)
erythroid cells Normoblast
2. Lymphopoiesis - Orthrochromic Normoblast
- B-cell - Reticulocyte
- B-cell growth factor - Erythrocyte
- T-cell - IL-2
Rubriblastic
MEGAKARYOPOIESIS
- Rubriblast
- Platelets came from megakaryocytes - Prorubricyte
- Involved in hemostasis and thrombus - Rubricyte
development - Metarubricyte
- Earlier influences include GM-CSF, - Reticulocyte
IL-3, IL-6, IL-11, kit ligand and EPO - Erythrocyte

Erythroblastic
 - Proerythroblast
- Basophilic Erythroblast
- Polychromic Erythroblast
- Orthochromic Erythroblast
- Reticulocyte
- Erythrocyte
MATURATION PROCESS
1. Erythroid Progenitors
- BFU-E (Burst Forming
Unit-Erythroid) 1 week
to mature
- CFU-E (Colony Forming
Unit-Erythroid) 1 week also towards
becoming
pronormoblast
- Approximately 6 days for
precursors to mature
- It will take approximately 18-21
days to produce a RBC
MATURATION SEQUENCE
1. Pronormoblast
- Nucleus: High (N8:C1),
round to oval with 1 or 2 nucleoli
- Cytoplasm: Quite blue (ER),
Golgi complex may be visible
- Division: Mitosis
- Location: in Bone Marrow
- Size: 18-20 um
- Cellular Activity: Accumulate
components for HB production and
globin production begins
- Length of time: Slightly more than
24hrs
2. Basophilic Normoblast
- Nucleus: Ratio (6:1), chromatin begins
to condense, staining reaction is deep
purple red
- Cytoplasm: deep blue
- Division: Mitosis
- Location: only in BM
- Size 16um
- Cellular Activity: Detectable HB
synthesis occurs
- Length of time: Slightly more than 24
hrs.
the
3. Polychromatic (Polychromatophilic)
Normoblast
- Nucleus: Ratio (4:1), chromatin
condensation, no nucleoli
- Cytoplasm: 1st stage Redness/pink is
associated with HB and concurrent
decrease of RNA; Mixed pink and blue
(Murky Gray-Blue)
- Division: Mitosis (Last stage capable)
- Location: in BM
- Size: 13um
- Cellular Activity: HB synthesis in
increasing and the accumulation is
visible in the cytoplasm; Organelles still
present
- Length of time: 30hrs.
4. Orthochromic Normoblast
- Nucleus: Completely condensed or
nearly so; Low NC ratio 1:2
 - Cytoplasm:
Pink-orange/salmon pink color (nearly
complete Hb production)
- Division: None
- Location: in BM
- Size: 8um
- Cellular Activity: Continuation of HB
production; Nucleus is ejected —>
Howell Jolly (HJ) bodies
- Length of time: approximately 48 hrs.
- Cytoplasm: Mature cells are biconcave
discs; Salmon-pink in color when
stained with a central area
(concavity)
- Division: Can’t Divide
- Location & Length: Active approximately
120 days in the circulation
- Size: 6-8um
- Shape: Biconcave
- Cellular Activity: Delivers Oxygen to
issues, releases it, and returns to the
lung to be re-oxygenated.

5. Polychromatophilic Erythrocyte
(Retyculocyte)
- Nucleus: None
- Cytoplasm: Predominantly the color of
HB
- Division: None
- Location: in BM for 1 - 2 days then
peripheral blood 1 day
- Size: 8um
- Cellular Activity: Completes the HB
production and endoribonuclease
digests the ribosomes
- Length of time: 2-3 days

6. Erythrocytes
- Nucleus: None