HEMATOLOGY

DIAGNOSTIC EXAM
MAGIGING RMT!!!
#1 MGA MUST KNOW!!

1/3 OF THE TOTAL PLATELETS (30-35%) ARE SEQUESTERED IN THE SLPEEN

2/3 OF THE TOTAL PLATELETS (70-75%) ARE FOUND IN THE CIRCULATION

ABOUT 25-30% RBCs ARE STORED IN THE SPLEEN

MGA MUST KNOW

CULLING
-DESTRUCTION OF THE SENESCENT RBCS THAT WILL LEAD TO EXTRAVASCULAR
HEMOLYSIS.
-RBCs ARE ENGULFED BY THE SPLEENIC MACROPHAGES.

PITTING
-THE PROCESS OF REMOVING INCLUSION BODIES FOUND ON THE RBCS
- IT WILL LEAD TO BITE CELL FORMATION
- COMMONLY ASSOCIATED IN CASES OF G6PD
 # 2 MGA MUST KNOW

Capillary tube:
 Length: 7-7.5cm or 70-75 mm

 Bore: 1-1.2 mm in diameter

 Plug: 4 to 6 mm long (seal the capillary tubes at the end of the tube with the colored ring)

 Two types: With red band (with anticoagulant heparin) and With blue or green band (without anticoagulant)

 Can hold up to 0.05mL of blood

 5cm filled portion with blood (2/3 filled portion)
 #3 MUST KNOW!
Fibrinogen group
(Thrombin Sensitive group)
♥ Factor I, V, VIII, XIII
♥ Consumed during coagulation
♥ Absent in serum
♥ Not absorbed by barium sulfate or
aluminum hydroxide
♥ Increases in inflammation,
pregnancy, stress and fear, oral
contraceptives
♥ The largest group
Prothrombin Group
(Vitamin K dependent Group)
♥ Factor VII, IX, X, II
♥ Requires vitamin K
♥ Not consumed during coagulation
♥ Absorbed by barium sulfate or
aluminum hydroxide
♥ Heat stable
♥ Increases in pregnancy, and oral
contraceptives
♥ Decreased in oral anticoagulants
Contact Group
♥ Factor XII, XI, HMKW, PK
♥ Not consumed during coagulation
♥ Not absorbed by barium sulfate or
aluminum hydroxide
♥ They are Stable
♥ They are so named “contact factors”
because they are activated by contact
with negatively charged foreign
surfaces
#5 MUST KNOW

1. Parameters that are directly obtained from histogram – WBC count, RBC count, Hemoglobin

2. Parameters that are derived from RBC Histogram – MCV and RDW

3. Parameters that are derived from platelet Histogram- MPV and PDW / MPV and platelet count

4. Parameters that are computed = Hematocrit, MCH, MCHC

5. THREE PART DIFFERENTIAL – Lymphocytes, Monocytes, and Granulocytes

6. FIVE PART DIFFERENTIAL – Neutrophils, Lymphocytes, Monocytes, Eosinophils, and Basophils

7. Ohm’s law: voltage = current x resistance

#7 MGA MUST KNOW

FAB ALL DESCRIPTION

SUBTYPE
L1 →Lymphoblasts are small and homogenous, varies little in size
→Scanty cytoplasm and inconspicuous nucleoli; nucleus is round and
irregular/indistinct in shape; have high N:C ratio
→Most common CHILDHOOD ALL with best prognosis
L2 →Lymphoblasts are large and heterogenous, variable in size
→Abundant, basophilic cytoplasm, and the nuclei are often clefted with
nucleoli present
→Adult type ALL
L3 → Burkitt-like/type
→Lymphoblasts are large, homogenous and vacuolated
→Rarest subclass, can be found in both children and adult
→Poor prognosis
#12 MUST KNOW

♥ NORMAL MYELOID TO ERYTHROID RATIO 2:1, 3:1, 4:1

♥ Once the differential (500-1000 COUNT) is completed in a bone marrow sample, the
myeloid-to-erythroid (M:E) ratio is computed from the total myeloid to the total nucleated
erythroid cell stages.

♥ Excluded from the M:E ratio are lymphocyte and its precursors, plasma cells, monocytes
and its precursors, histiocytes, non-nucleated erythrocytes, and nonhematopoietic stromal
cells
HUWAG MACONFUSE!!!

MARROW CELLULARITY
- Ratio of marrow cells to yellow marrow (fat) and typically decreases with age
MARROW TYPE DESCRIPTION
Red/Active marrow Hematopoietically active marrow consisting of
the developing blood cells and their
progenitors
Yellow /Inactive marrow Hematopoietically inactive marrow composed
primarily of adipocytes (fat cells), with
undifferentiated mesenchymal cells and
macrophages
HUWAG MALITO!!

• RETROGRESSION
- The process of replacing the active marrow by adipocytes during development
#18 MUST KNOW!!
#20 MGA MUST KNOW!!
#21 MUST KNOW

PARAMETER SHIFT TO THE LEFT SHIFT TO THE RIGHT

PH Increase Decrease

PCO2 Decrease Increase

2, 3- DPG Decease Increase

TEMPERATURE Decrease Increase

HB AFFINITY FOR O2 Increase Decrease

• SHIFT TO THE LEFT

=DON’T LEFT the Fetus (fetal hb) inside My (Myoglobin) Car(Carbon monoxide)

• SHIFT TO THE RIGHT

= Ha HA! (High altitude, Hypoxia, Anemia)
 WBC  1-3% Acetic acid
 1% HCL
#25 MUST KNOW 

1% Ammonium oxalate
Turk’s diluting fluid
(Glacial acetic acid + Crystal/Gentian violet)

EOSINOPHIL  Pilot’s fluid

 Randolph’s fluid
 Manner’s fluid
 Tannen’s fluid
 Hinkelman’s fluid

RBC  NSS
 Dacies or Formol citrate (BEST)
 3.8 % Sodium citrate
 Hayem’s
 Toisson’s
 Grower’s
 Bethell’s

PLATELET  1% Ammonium oxalate

 Direct methods for platelet counting (Neubauer) = Brecher-
Cronkite, Leake and Guy, Unopette, Reese Ecker

Hemoglobin  Drabkin’s reagent

#29 MUST KNOW

GRANULES MATURATIONAL STAGE/S PRODUCED

Primary (Non-specific or azurophilic) Promyelocyte stage
Secondary (Specific) Myelocyte and Metamyelocyte stages
Tertiary (Gelatinase) Metamyelocyte and Band cell stages
Secretory vesicle or granules Band cell and Segmenter(mature) stages
#30 MUST KNOW
#35 MUST KNOW!

IMMATURE CELLS MATURE CELLS

Cell Is large Cell becomes smaller

Nucleoli is present Nucleoli absent

Chromatin fine and delicate Chromatin coarsed and clumped

Nucleus round Nucleus round, lobulated or segmented

Cytoplasm dark blue (rich in RNA) Cytoplasm light blue (less RNA)

High N:C ratio Low N:C ratio

AS THE CELL MATURES:

1. DECREASE IN SIZE
2. DECREASE N:C RATIO
3. DECREASE NUCLEAR SIZE
4. LOSS OF BASOPHILIA
5. LOSS OF NUCLEOLI
6. FROM FINE AND DELICATE CHROMATIN TO CCC
(COARSED, CLUMPED CONDENSED)

chromatin pattern = best way in differentiating immature from mature cell (WBC and RBC)
#42 MUST KNOW

ENDOMITOSIS!!
- WITH NUCLEAR DIVISION
- NO CYTOPLASMIC DIVISION
- NO CYTOKINESIS
- NO TELOPHASE
- NO DAUGTHER CELLS CREATED
 Microtubules  8 to 20 tubules per platelet
 Major responsible for platelet disc shape. It maintains platelet disc
shape.
 On cross section, microtubules are cylindrical, with a diameter of 25 nm.

#43 MUST KNOW

The circumferential microtubules could be a single spiral tubule
 Disassemble at refrigerator temperature or when platelets are treated
with colchicine
 When microtubules disassemble in the cold, platelets become round, but
on warming to 37° C, they recover their original disc shape

Microfilaments  Composed of actin and myosin or actomyosin (thrombosthenin)

 Actin and myosin, which upon stimulation of platelet will interact to form
actomyosin (thrombosthenin), a contractile protein for platelet contraction
 Actin is present throughout the platelet cytoplasm, constituting 20% to 30%
of platelet protein
 Resting platelet actin is globular and amorphous
 Activated platelet actin becomes filamentous and contractile

Intermediate  Composed of desmin and vimentin

filaments
 They are ropelike polymers 8 to 12 nm in diameter
 Intermediate filaments connect with actin and the tubules, maintaining the
platelet shape
#45 MUST KNOW
ISBB DIAGNOSTIC
EXAM
MAGIGING RMT!!!
#1 MGA MUST KNOW!

CYTOKINES
• CYTOKINES ARE CHEMICAL MESSENGERS

• CYTOKINES ORCHESTRATE BOTH NATURAL AND ADAPTIVE IMMUNITY

• PRODUCED BY MANY CELLS BUT THE MAIN PRODUCERS ARE T-CELLS , MONOCYTES,
AND MACROPHAGES
• CYTOKINES ARE NOT PRODUCED BY B-CELLS

CYTOKINES PRODUCED BY T AND NK CELLS = LYMPHOKINES

CYTOKINES PRODUCED BY MONO/MACRO = MONOKINES
#2 MUST KNOW
#3 MUST KNOW
#6 MUST KNOW

 Bga = Represents HLA B-7

 Bgb = Represents HLA B-17

 Bgc = Represents HLA A-28

BENNETH GOODSPEED ANTIGENS

#7 MUST KNOW

ACQUIRED B PHENOMENON

*NAHALUAN NG B antigen ANG RBC MO!

* TRUE PHENOTYPE: Blood type A

*MISTAKEN AS: Blood type AB (Psuedo AB)

*ASSOCIATED WITH EPIC ( E.coli 086, P.vulgaris, Intestinal obstruction, Colorectal cancer)
#8 MUST
KNOW
 #14 MUST KNOW
ADJUVANT COMPOSITION

Aluminum One or more of the following: amorphous aluminum hydroxyphosphate sulfate (AAHS), aluminum hydroxide,
aluminum phosphate, potassium aluminum sulfate (Alum)

AS01B Monophosphoryl lipid A (MPL) and QS-21, a natural compound extracted from the Chilean soapbark tree, combined in
a liposomal formulation
AS04 Monophosphoryl lipid A (MPL) + aluminum salt
CpG 1018 Cytosine phosphoguanine (CpG), a synthetic form of DNA that mimics bacterial and viral genetic material

Matrix-MTM Saponins derived from the soapbark tree (Quillaja saponaria Molina)

MF59 Saponins derived from the soapbark tree (Quillaja saponaria Molina)
#14 MUST KNOW

THE FIRST ADJUVANT ( NOT FOR HUMAN USE!!!)

-The first such adjuvant was discovered by Freund in the 1930s and is known as Freund’s
complete adjuvant (FCA).
-FCA is a powerful adjuvant containing killed mycobacteria. It has been used in
animal studies, but is not suitable for use in humans because it produces abscesses and scar
formation at the site of inoculation

-Freund’s complete adjuvant (mineral oil, emulsifier, and killed mycobacterium)

#17 MGA MUST
KNOW
DR -2
= Two (too) Good to have MuScle
= Good Pasteur and Multiple Sclerosis

DR-4
= Si Juven ay isang Malaking Pegasus (pemphigus) na may Rheumatoid arthritis
= Juvenile DM, Giant cell arthritis, Pemphigus, RA
NOTE: RA pwede gawing R4 (bawasan lang ng ISANG paa yung letter A xD)

DR-3
= TATLONG BESES SaSaGAD!!
= SLE, Sjogren, Gluten enteropathy, Addison, Dermatitis Herpetiformis
NOTE: SLE pwede gawing SL3 (ang E pag binaliktad ay parang 3)
DR-5
=Hashi CLLeaN that Scary Pussy (posi)
=Hashimoto, CLL, Nepropathy, Scleroderma, KaPOSI (pussy) sarcoma
#17 MUST KNOW

 HLA B8 = Confers additional protection and increased resistance to HIV infection

 Celiac disease is associated with HLA DQ2 and DQ8
 Type 1 diabetes mellitus is associated with HLA DQ2 and DQ8
 Psoriasis vulgaris is associated with HLA-Cw6
 Gold-induced nephropathy, Kaposi sarcoma, & Chronic lymphatic leukemia are associated
with HLA-DR5
#23 MUST KNOW

• Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-

related morbidity and mortality
# 25 MUST KNOW

Type of blood donation Hemogloin Hematocrit

Allogeneic donation At least 12.5 g/dl At least 38%

Directed donation At least 12.5 g/dl At least 38%

Autologous donation At least 11 g/dl At least 33%

Double RBC pheresis Not indicated At least 40%

#26 MUST KNOW

• CDC criteria for Western Blot to confirm as positive: a result should be reported as
positive if at least two of the following three bands are present: p24, gp41, and
gp120/gp160.
#28 MUST KNOW

“TORCH or TORCHeS”
 CONGENITAL INFECTIONS!!!!
 NOT STD OR NOT HEREDITARY CONDITION
 Transmitted from mother to fetus (vertical or perinatal)

T= Toxoplasmosis
R = Rubella (3 DAYS MEASLE / GERMAN MEASLES)
C= CMV (most common)
H= Hepatitis, Herpes, HIV
S = Syphilis
 #34 MUST KNOW

EXCHANGE TRANSFUSION

*Primary indication: Removal of Unconjugated bilirubin

*Sample used for crossmatching: Mother’s serum (preferred), Infant’s serum, or Cord RBCs

*DONOR RBC Should be:

Fresh as possible (less than 5 or 7days old), Group O negative or group specific, CMV negative, Hgb S
negative, and irradiated
#37 MUST KNOW

GENE GLYCOSYLTRANSFERASE IMMUNODOMINANT SUGAR Antigen

H L-fucosyltransferase L-fucose H

A N-acetylgalactosyltransferase N-acetyl-D-galactosamine A

B D-Galactosyltransferase D-galactose B

AB N-acetylgalactosyltransferase N-acetyl-D-Galactosamine AB

D-Galactosyltransferase D-galactose

O -- -- Unchanged
#40 MGA MUST KNOW

CD4: CD8 RATIO

*NORMAL = 2:1
*In AIDS = 1:2 or 0.5:1

Normal CD 4+ COUNT = 500-1300 cells/ ul (or 450-1500 cells/ul – Steven’s)

#41
#41 MGA MUST KNOW

HYBRIDOMA Formation of a hybridoma in monoclonal antibody production. A mouse is

TECHNOLOGY immunized, and spleen cells (plasma or B cells) are removed. These cells are fused
with non-secreting myeloma cells and then plated in a restrictive medium. Only the
hybridoma cells will grow in this medium, where they synthesize and secrete a
monoclonal immunoglobulin specific for a single determinant on an antigen.
HYBRIDOMA An immortal cell capable of producing indefinite sequence of nucleotides
Animal used Mouse
Surfactant PEG (Polyethylene glycol)- for fusion of plasma cell with myeloma cell
Selective medium Medium with HAT (Hypoxanthine, aminopterin, and thymidine)
Monoclonal antibodies purified antibodies cloned from a single cell. These antibodies exhibit exceptional purity
and specificity and are able to recognize and bind to a specific antigen.
 #47 MUST KNOW
S>E>C>E>S (ANTIGEN MUNA BAGO ANTIBODY)

MARKER COMMENT
HBsAg First serological maker to appear
(Surface) Present in active, acute, and chronic infection
Important marker in Screening blood donors

HBeAg Marker for active replication of the virus

(Envelope) Marker for high degree of infectivity

HbcAg Not detectable in serum (not tested in serology)

(Core) Detected only through biopsy of the infected liver
 HEPATITIS B MARKERS
MARKER COMMENT

Anti-HBc IgM (HBcAb IgM) Indicator of current, recent, or acute infection

Marker of “Core Window” period

Anti-HBc IgG (HBcAb IgG) Indicator of chronic or past infection

Serves as a lifelong marker of HBV infection

Anti-HBc Total Measure of both IgM and IgG

Indicates previous or ongoing infection

Anti-HBe (HBeAb) Marker of convalescence and favorable(good) prognosis

A negative Anti-HBe test is associated with ineffective therapy
A positive Anti-HBe test indicates that the current therapy is effective

Anti-HBs (HBsAb) It is the Serologic marker of recovery and immunity

Persists for years and provide protective immunity
 TESTS RESULTS INTERPRETATION
HBsAg Negative Susceptible

Anti-HBc (total) Negative

Anti-HBs Negative
HBsAg Negative Immune because of natural infection

Anti-HBc Positive

Anti-HBs Positive
HBsAg Negative Immune because of hepatitis B vaccination

Anti-HBc (total) Negative

Anti-HBs Positive
HBsAg Positive Chronically infected

Anti-HBc (total) Positive

IgM anti-HBc Negative

Anti-HBs Negative

HBsAg Negative 1. Might be recovering from acute HBV infection.

Anti-HBc Positive 2. Might be distantly immune and test not sensitive enough to detect very low level of anti-HBs
in serum.
Anti-HBs Negative
3. Might be susceptible with a false-positive anti-HBc.

4. Might be undetectable level of HBsAg present in the serum and the person is actually
chronically infected.
END