 Mature

 Gravid – Contains ova ready to release in the system

 Handling feces as the main sample
 Identification of different parasites
 The work of junior medtechs. (Hematology and Clinical Microscopy)
BASIC TERMINOLOGIES
PARASITE
 Any organism that depends on another organism for shelter and
environment/nourishment
 Garapata, ticks
HOST
 Organism that supports the parasite
ECTOPARASITE
 Definitive Host
 Thrive externally on a host (Outside the host); Grossly macroscopic
 Harbors the adult stage
 Infestation
 Intermediate Host
 Harbors the larval/asexual form
 Infection stage

EOSINOPHILIA – Bigger than RBC

 Increase in eosinophil count (WBC)
 Charcot-Leyden crystals
 Possess granules to kill parasites
 Count – in the blood
 Slightly larger than neutrophil
 Combats bigger organisms
 Found mostly in feces and sputum
 Parasites can migrate
TAENIA SPECIES
 Taenia solium - host is pork (pigs); fecal contamination (from ova)
 There is cysticercosis when it reaches the brain
(cerebrum and cerebellum) (larva); Acquired through
ingestion of tapeworm
 CYSTICERCOSIS – In clumps/clusters, when
destroyed, larva is found
 Creates seizures when it inhabits the brain (Other nervous
 INCREASE – hypersensitivity or parasitosis; If infected with a parasite
system conditions)
CHARCOT-LEYDEN CRYSTALS
 Taenia saginata – host is beef (cows)
 Remnant of the granules
 Creates seizures when it inhabits the brain (Taenia solium)
 If found in feces, eosinophil count is high
 TAENIASIS – Ingestion of infected pork, undercook; Disease caused by  Sharp in appearance
 PHYLUM OF TAENIA – PLATYHELMINTHES

MAJOR GROUPS OF IMPORTANT PROTOZOANS

A. PROTOZOANS
 Phylum composed of simplest organism
 Dark part of the body contains the ova which is released to the body  AMOEBA
through a hole at the side of the body  Have locomotor structures
MODE OF TRANSMISSION  Entamoeba histolytica – most common amoeba
 How a parasite successfully enters a susceptible host  Trophozoite and Cyst: Parts of the life cycle; Stages
 There are lots of types  Trophozoite
 Airborne  Vegetative stage
 Motile stage
 Waterborne
 Watery feces
 Vector
 Cyst
 E.g. Mosquito
 Firm stool
 Skin Penetration
 Usually found in firm
 Larval stool
 Sexual Contact  Has a locomotive
 Infestation – Direct or Indirect structure
 E.g. Lice ENCYSTATION – trophozoites ➝ cyst
 Formation of cyst from trophozoite
 Pathogenic parasites – Disease-causing
EXYSTATION – reverse process
 Non-pathogenic parasites – Do not harm the host
 Cyst ➝ trophozoite
ENDOPARASITE
 Found inside the body of the host
 Infection
PROGLOTTIDS – Body of the hookworm
 3 Types
 Immature
  FLAGELLATES – Has flagella for movement
 With whip-like structures
 All demonstrate trophozoites form; No cyst form
 Not all are capable of encystation
 Trichomonas spp. – very motile
 T. vaginalis (Trichomonas vaginalis) –
common
 T. hominis
 T. tenax
 D. fragilis – Dientamoeba fragilis
 Thrive in small intestine
 Hemoflagellates – Blood; Both common diseases are
vector borne and are found in African countries
 Trypanosoma cruzi – Most common;
causes Chagas disease via kissing bug
(bites the lips); causes fever and body
ache
 Leishmania donovani – causes Kala-Azar
(black fever) which is transmitted via
sandflies; causes infections, spleen
fever, and anemia
 CILIATES
 Balantidium coli
 DISEASE: Balantidiasis
 Attributed to pigs
 Pigs
 Has cilia; Very Motile
 Roundworms
 Phylum Nemathelminthes
 Adult Worms: Tapered, Cylindrical bodies with an
esophagus and longitudinal muscles
 Dioecius = They have separate sexes (male and female
adult groups)
 Male = Has a posterior curve; Smaller and
shorter
 Female = Larger because it bears the ova
 Monoecious = Protozoans
 Most common hookworm = Necator americanus
THE UNHOLY THREE
 If one is found, the other two are expected to be seen as well
 Common in school age children
 MODE OF TRANSMISSION – Larval skin penetration (walking
around barefoot)
 Larva = Infected stages of the parasites (also ova)

1 – Hookworm Egg
 Has a clear cytoplasm
 Cannot be identified in ova stage; Differentiated in adult stage
2- Trichuris trichuria egg
 Football shaped
3 – Ascaris lumbricoides (Decorticated egg)
 SPOROZOANS  Most common
 Plasmodium spp. – most common; causes malaria  Shiny and smooth
 Plasmodium falciparum – commonly C. TREMATODES
found in the Philippines  Flukes or flukeworm
 P. vivax  2 Orders: Monogenia and Digenia
 P. ovale  Flat, leaf-like and hermaphroditic (Except schistosoma spp.)
 P. malariae  HERMAPHRODITIC – Contains both male and
 P. knowlesi female reproductive organs
 Vector borne  2 intermediate hosts
 Recently added because of 1. Snail
the involvement of humans; 2. Varies (Plants, snails, crabs)
Comes from monkeys  SCHISTOSOMES

S. haematobium urine = Goes to the urinary bladder (Lacerates mucosa to the

urinary system; Hematuria)
 Spine – Causes damage to the mucosa

Red Blood Cells – If not enlarged but infected – P. malariae or Falciparum spp.
 Enlargement is caused by P. ovale/vivax (infected)
B. NEMATODES

Copulation of the adult male and adult female schistosome (Flukeworm)

 Male – Has a spicule (penis of the male flukeworm)
 Female – Larger because it bears the ova
 Possesses the bursa
 BURSA – Where the male worm
resides
D. CESTODES
  Tapeworms (Taenia)  Black – Upper GIT bleeding (smaller
 Dwell in small intestines intestine)
 Parts: Scolex (Head), Neck and several proglottids  Occult (Hidden) Blood – More likely a sign of
(Segments) other GIT disorders
 Abdominal pain, dyspepsia, anorexia, nausea, diarrhea  Stercobilin – Produces the color of the stool
 TREATMENT: Get rid of the scolex (head) of the tapeworm  Stool Consistency; Normal pH = 7.0-7.5 (feces)
in order to stop regeneration  Well-formed – If when poked by
applicator stick, there is pressure
 Semi-formed – If poked by
applicator stick, it is smooth (soft)
 Mushy or liquid (watery)
 Predictor for possible
stages of parasite in
the sample (All)
 If liquid = trophozoite (motil)
 If formed = cyst
 If frothy = cyst
 Frothy – whip cream-
like consistency
 FECAL OCCULT BLOOD TEST – Identifies if
there is blood in the feces/stool (hidden)
 How to test the stool consistency
 Feces is smeared in the
reagent and wait for a
change in the color =
blue
 MACROSCOPIC
 What is seen in the laboratory
results
 Light-brown – color
 Soft – consistency
 Odor does not need to be reported
 Method of Stool Examination – Microscopic Examination
 Identifying the size, done in research lab
DIAGNOSTIC PARASITOLOGY
 For identification of helminth eggs and larvae
 Tests that are important for diagnosis of parasite infection
as well as protozoan’s cyst and trophozoites
A. STOOL EXAMINATION: FECALYSIS
 Parasite micrometry: Use a calibrated Ocular
 Collection of Fecal Specimens Micrometer
 Thumb-sized stool specimen (2-4 grams)
 Placed in a dry, clean wide-mouth container
with tight-fitting lid (bacteria can give off gas)
 LABEL THE STOOL CONTAINER PROPERLY
 Sample must be free from any contamination

 Micrometer is placed in the ocular lens

 Not routinely done, usually for research
 Wet Mounts – Used for actual fecalysis
 Direct wet mount
 Fresh, very soft
or liquid stool
 Label must be placed on the body of the sample
container to avoid switching specimen; Not in  Detect/Identify
the cap = Possibility of switching/loosing motile
sample trophozoites
 Must be read within 1-2 hours  1-2 drops of water stool →
 CONTAMINATION – There must be no water, Glass slide → Cover slip →
diaper contamination, and urine contamination; Microscope
Reflect the sample
 Would lead to a false result
 Defer for 1 week if patient took anti-diarrheal
medication, radio-opaque compounds, or oily
laxatives
 Stool specimen shouldn’t be frozen
 Unpreserved specimen shouldn’t be stored at
room temperature for more than 2 hours
 10% FORMALIN – Preserves stool
specimen for 24 hours
(Morphology of the ova);  Saline Wet Mount – Indirect;
Preservative Depends on what is placed
 Method of Stool Examination – Gross Examination  Norma Saline
 Stool Color Solution (NSS) =
 Brown – Norma; Because bilirubin Fecal emulsion;
turns to stercobilin Adjust light of
 Bright Red Blood – Bleeding microscope –
hemorrhoids/lower GIT bleeding Presence of
 Bloody Mucus (Loose or Liquid) – protozoan
Suggestive of ulceration in large trophozoites
intestines
  NO LUBRICANT – Lubricant will interfere with
the swab (mucosa)
 Mucosa is swabbed and then smeared on the glass slide

 Iodine Wet Method

 Lugol’s Iodine
 Highlights the
details the
protozoan cyst
 Iodine is brown
 IF NONE; REPORTING = No
parasite seen
What is Clinical Chemistry?
 It is the branch of medical science involved in the analysis of biological
materials, usually body fluids, to provide diagnostic results on the state
of human body
BASIC ANALYTICAL TECHNIQUES
1. SPECTROPHOTOMETRY
B. COLLETION OF BLOOD SPECIMENS  The measurement of the intensity of light at selected
 Detects Plasmodium, filanal worms, Trypanosoma, and wavelengths
Leishmania  A useful analytical tool to determine the concentration of
 Definite time of collection = Parasites exhibit periodicity colored material in a solution
(Some are nocturnal)
 Palmar surface of the distal segment of the third (middle)
or fourth (ring) finger
 SKIN PUNCTURE – Use a lancet

 Uses prisms or gratings to isolate a narrow range of

wavelength of light

C. COLLECTION OF SCOTCH TAPE SWAB

 Useful in cases of Enterobiasis = Intense itchiness in
perianal area
 Where adult female deposits ova because it is
airy
 Usually occurs every morning
 SWAB
 Pre-cut scotch tape  It employs two kinds of light:
 Tongue depressor  VISIBLE LIGHT = 340-700nm
 Glass slide  INVISIBLE LIGHT
 Ultraviolet Light = < 340nm
 Infrared Light = > 700
BASIC PARTS OF A SPECTROPHOTOMETER

1 & 2 – Loop tape over end of slide to expose gummed surface

3 – Touch gummed surface several times to perianal region
4 – Smooth tape on side, after apply drop of toluol or I2 in xylo
2. NEPHELOMETRY
 ENTEROBIUS VERMICULARIS – Used in scotch tape swab
 Light scattered by an unknown substance is measured at
 FAMILIAL = Easily obtained within the family
right angles
 Depends on wavelength and particle size
3. TURBIDIMETRY
 Measure the amount of light blocked (absorbance) by a
suspension of particles
 Depends on particle size and concentration
4. ELECTROPHORESIS
 Charged molecules move at different rates when pulled
D. COLLECTION VIA VAGINAL SWABS through an electrical field
 Diagnosis of Trichomonas vaginalis infection (motile)  Cations (Positively charged ions) will move to the cathode
 BD Affirm VPIII Ambient Temperature Transport System (negative electrode)
(ATTS)  Anions (Negatively charged ions) will move to the anode
 Molecular system for detection of vaginitis (positive electrode)
 Speculum without lubricant is used – inserted in vagina and
opens once inside
  FUNCTION
 Act as energy source (next to glucose)
 Important constituent of cell membrane
 Triglyceride
 Cholesterol
 Lipoproteins (HDL, LDL, VLDL, chylomicrons)
C. KIDNEY (RENAL) FUNCTION TESTS
 KIDNEYS
 Paired organs considered as the body’s major
“waste sweeper”
1. Creatinine
 Waste product of
muscle metabolism
2. Blood Urea Nitrogen (BUN)
 Waste product of
protein catabolism
5. FLAME EMISSION SPECTROPHOTOMETRY  90% is excreted in
 Measures light emitted by excited atoms urine
 Certain elements give off a characteristic light after 3. Glomerular Filtration Rate (GFR)
 Used to direct/check
excited atoms return to ground state
 SODIUM – Intense yellow flame how well the kidneys
 POTASSIUM – Violet flame are working
 Estimates how much
 CALCIUM – Brick red flame
blood passes through
6. ATOMIC ABSORPTION SPECTROPHOTOMETRY
the glomeruli each
 Measures light absorbed by ground state atoms
minute
 Routinely used to measure the concentrations of trace D. LIVER FUNCTION TESTS
metals
 LIVER
 Hollow cathode lamp is the usual light source employed  Organ responsible for the synthesis of many
CLINICAL CHEMISTRY ASSAYS organic substances
A. BLOOD GLUCOSE  Detoxifies the body against noxious substances
 Glucose is the primary source of energy for humans 1. Bilirubin
 Normally increase after eating 2. Liver Enzyme Tests
 SHORT TERM STORAGE SITES: Liver and 3. Total Serum Protein Tests
skeletal muscles 4. Prothrombin Time
 LONG TERM STORAGE SITES: Adipose tissues E. CARDIAC FUNCTION TESTS
 May appear in urine if renal threshold (160-180 mg/dL)  To evaluate whether one’s heat is healthy or not
has been exceeded  Patients who experienced episodes of myocardial
 CLINICAL SIGNIFICANCE: Diabetes infarction as manifested by chest pain
 DIABETES MELLITUS 1. Troponin Test
1. Defect in the beta cells of the  Most sensitive and specific test
pancreas for myocardial damage
2. Leads to a decrease in the 2. Myoglobin
production of insulin (A hormone  Along with troponin, ordered as a
necessary to maximize the cardiac biomarker to help diagnose
utilization of glucose for energy or rule out a heart attack
production)  Increase later than troponin but
3. SIGNS & SYMPTOMS: 3P’s not specific and will not stay
 POLYURIA – excessive elevated for long as troponin
urination 3. Other cardiac enzymes:
 POLYDIPSIA –  Creatine Kinase (CK-MB)
excessive thirst  Aspartate Aminotransferase (AST)
 POLYPHAGIA –  Lactate Dehydrogenase (LDH)
excessive eating F. TUMOR MARKERS (SPECIAL CHEMISTRY TESTS)
 TESTS TO BE DONE:  A biomarker indicative of an inherent cancerous condition
 Fasting Blood Sugar (FBS) 1. AFP (Alpha-Fetuprotein) –
1. 6-8 hours fasting prior to blood hepatocellular carcinoma
collection 2. CEA (Carcinoembryonic Antigen) –
 Random Blood Sugar (RBS) gastrointestinal cancer
 Oral Glucose Tolerance Test (OGTT) 3. PSA (Prostate Specific Antigen) –
1. For pregnant women to rule out prostate cancer
cases of gestational diabetes 4. hCG (human Chorionic Gonadotropin)
 2-Hour Post-Prandial Blood Sugar (2HPPBS) – gestational trophoblastic disease
 Glycosylated Hemoglobin HBA, C) 5. NSE (Neuron Specific Enolase) –
1. To determine if diabetes has been neuroendocrine tumor
existing for several months already 6. CA 125 – ovarian cancer
(long-term diabetes) 7. CA 19-9 – pancreatic cancer
2. It reflects the average blood 8. CA !5-3 – breast cancer
glucose over a three-month period 9. Calcitonin – medullary thyroid
3. Measured through affinity carcinoma
chromatography 10. Desmin – smooth muscle
B. BLOOD LIPID PROFILE carcinoma
 LIPIDS
 Organic substances characterized by their
general insolubility in water and solubility in IMMUNOLOGY
organic solvents  The study of all aspects of the immune system, including its structure
 Regarded as “fat” and function
 1 gram = 9kcal of heat
  It deals with the response of an organism to antigenic challenge and 4. Epsilon (IgE)
its recognition of what is self and non-self  Increased during hypersensitivity and
 NON-SELF – foreign substance; stimulates antigenic parasitism
response 5. Delta (IgD)
 Principle  Found on the B-cell surface (outside the B-cell)
SEROLOGY
 A division of immunology that specializes in the laboratory detection
and measurement of antigens and antibodies
 Laboratory; Do various tests in the laboratory
 Receive samples that are the liquid portion of the clotted blood (serum)
 Plasma – Anti-coagulated blood (liquid portion)
 Serum – Clotted blood (liquid portion) BODY LINES OF DEFENSE
IMMUNITY Specific Defense
 The ability of an organism to resist a particular infection or toxin by Nonspecific Defense Mechanisms Mechanisms (Immune
the action of specific antibodies or sensitized white blood cells System)
 Needs to be built through time First Line of Defense Second Line of Defense Third Line of Defense
 Child needs to go out more often to build resistance from various  Skin  Phagocytic  Lymphocytes
infections or diseases  Mucous white blood  Antibodies
 Immunobuilders – Vitamins & minerals; Helps you improve membranes cells
your immune system  Secretions  Antimicrobial
ANTIGENS (Ag) of skin and proteins
 A substance that is capable of stimulating an immune response mucous  The
 Any foreign substance (protein, virus) membranes inflammatory
 Agglutinogen (Ag) response
 EPITOPES – Binding site for antibody (different/various) FIRST LINE OF DEFENSE
ANTIBODIES  Skin
 Proteins made by plasma cells in response to an antigen  Must be intact; Open wound is a great avenue for the
 Comes from our body bacteria to go in
 Counteracts antigens  Except larval skin penetration
 Y-shaped  Mucous Membranes – Cilia; Mucus; Sebum (secretion)
Transformed B-cells → Plasma cells → Proteins →Antibodies  Has antibacterial properties
 Proteins – In general, they are specific to a type of disease  Normal Flora – Normal bacteria residing in our body
2 types of lymphocytes:  Stomach Acidity
 Named according to which body part they matured  Has chyme, which is made of HCl acid
 B-cell – Originated from the bone marrow  Some bacteria dies of too much acidity
 T-cell – Originated from the thymus  Tears – Has an enzyme Lysozyme, which has antibacterial properties
 2 main parts of the antibody that cleans the eyes
 The 2 portions of the immunoglobulin SECOND LINE OF DEFENSE – Cellular
 FAB (Fragment of Antigen Binding)  Phagocytes – Cells capable of phagocytosis
 Where the antigen binds/binding  Phagocytosis – Engulfing foreign substances or bacteria
site (debris, fragments)
 FC (Fragment of Crystallization)  Neutrophil – Most phagocytic
 Antibodies are capable of  Eosinophil
crystallization  Basophil
 Monocyte
 Granulocytes are all phagocytic
 WBC – Capable of creating phagocytosis; Biggest WBC is also a
phagocyte (except Lymphocytes)
 Antimicrobial Proteins – Secreted by other cells
 Inflammatory Response – Fever is included; Body wants to get rid of
foreign substance which is why it heats (Some bacteria cannot thrive
in high temperatures)
 Lymph Nodes – Also part of the immune system; Biggest
 5 types of antibodies lymph nodes are inside the upper thighs; Will inflame to
 IgM, IgG, IgA, IgD, IgE (GAMED) isolate the bacteria from a wound which would come in
 Ig – Immunoglobulin; (Letter Y in structure) the system of the body
1. Gamma (IgG) THIRD LINE OF DEFENSE
 Most common  Lymphocytes – B-cells; Memory cells; Has a lifespan of 20 years
 Occurs after an infection  Antibodies – Once infection is done, it becomes a memory cell
 Capable of crossing the placenta  Shingles – Reactivated chicken pox; Not contagious
 IgG is passed from the mother to the fetus in
 Vaccines – Should be infected at childhood; Develops antibodies
the womb
2. Alpha (IgA)  Measles
 Found in secretions in the body (saliva, feces)  Mumps
 2 types of alpha  Rubella
 Monomer (monomeric) – G, E, and TYPES OF IMMUNITY
D are monomeric
 Dimer (dimeric)
3. Mu (IgM)
 First
 Primary response to an infection
 Has the biggest structure
 Bigger structure – More efficient
in combatting the antigens
 The only pentameric antigen
 Pentameric – 5x larger than the
usual size
  Population  Plays an important role in the diagnosis of diseases using
 Farmed Immunity – Most of the people in the community different tests
has been vaccinated  IMMUNOASSAYS – Designed to detect the presence of either antigen
 Person not vaccinated becomes immune as well or antibody in the unknown sample
 Lagtime – Gives the body time to create the antibody  Enzyme Immunoassay (EIA)
 NATURAL  Radio Immunoassay (RIA)
 Active  Fluorescent Immunoassay (FIA)
 Body makes the antibody when there is an  Chemiluminiscent Immunoassay (CLIA)
infection IMMUNOLOGY AND SEROLOGY LABORATORY TESTS
 Acquired natural active immunity  Bacterial Agglutination Tests
 Passive  Agglutination/Agglutinate – Involves antigens; Clumps
 Body receives the antibodies  To determine bacterial infections associated with
 Accepts the antibody persistent fever
 Natura because it comes from a person  Makes use of bacterial antigens to detect the presence
 E.g. Transplacental transfer of immunoglobulin of antibodies (reagent)
gamma  Examples:
 COLOSTRUM – Yellowish  Widal Test – Typhoid fever (foodborne) – Salmonella typhi
substance in breastmilk that is rich  Quite similar to dengue; Coinfects with dengue
in immunoglobulins (from mother to  Usually misunderstood as dengue
womb)  Weil-Felix Test – typhus fever (flea borne) – Rickettsia
 ARTIFICIAL typhi
 Active  Antistreptolysin O (ASO)
 No sickness detected but is infected by the  Diagnostic test for streptococcal infections
vaccine (antibody)  Rapid Plasma Reagin (RPR)
 Not necessarily needed
 Non-specific test for syphilis (treatable at an early stage)
 Body produces antibody that is still part of the
 Treponema Pallidum Hemagluttination Assay (TPHA) Test
virus (antigenic)
 Specific test for syphilis
 E.g. MMR vaccines
 Hemagluttination
 Passive
 Reagent used contains blood
 Antibody itself is injected in the body
 Can use animal blood
 Body did not produce but antibody is introduced
 Cross Reactivity – Other viruses and bacteria can cause positive
to your body
results
 Has parts of the virus that cause the disease
(Not capable of creating disease because of  C-Reactive Protein (C-RP) Test and C3C Radioimmunodiffusion-Assay
the vaccine components) (RID) Test
IMMUNOLOGY-SEROLOGY  Non-specific marker for inflammation/infection
 Needs more volume of blood
 Tests for Heterophilic antibodies
 Infectious Mononucleosis (IM) – caused by Epstein-Barr
virus
 Paul-Bunnell Test – Presumptive Method
 Davidson Differential Test – Classical
1. Identify and quantifying Reference Method
 Antigens  Monospot Test – Qualitative test that detects
 Antibodies IM heterophilic antibodies in the sample
 Antibody Titer (Quantification)  Qualitative – sense or presence
 Hepa B (required): Normal value = 100 and  Quantitative – Quantitate number
above of antibodies and antigens
 Quantity of antibodies needed to combat the  Pregnancy Test
disease  Detects -human Chorionic Gonadotropin (-hCG)
 If quantity does not reach required amount =  Hormone produced by a developing embryo
BOOSTER  Sample is blood
2. Investigating problems or disorders with the immune system, such as  Results can be seen in 3-5 minutes
autoimmune diseases and immunodeficiency disorders  HIV Tests
 AUTOANTIBODY – Produces antibodies when the self and  HIV Rapid Test – Screening
non-self is no longer detected  Wait 10-15 minutes for the results
 Systematic Lupus Erythematosus (SLE)  Detects HIV 1 and 2
 Autoimmune disorder that is common in young  If REACTIVE (Positive), send to:
women  Western Blot Test – Confirmatory; For cross
 Cause is unknown; Research says that it is reactivity sent to RITM (When blood comes
attributed to UV rays, medicine, viruses, and from blood banks) and San Lazaro Hospital (If
stress positive or reactive)
 If you have antinuclear antibodies
 Multiple Sclerosis (MS)
 Affects the nervous system and makes you
wrinkly like a vegetable
 Rheumatoid Arthritis
 Psoriasis
 Skin disorder that has no treatment but is not  Hepatitis B Antigen Rapid Test (HBsAg surface rapid test)
contagious  Rapid qualitative screening test for the detection of
3. Determine organ, tissue, and fluid compatibility for transplantation hepatitis B infection
 Important
 E.g. Bone marrow transplant – Best donor is an identical
twin or a sibling
PRINCIPLES OF IMMUNOLOGIC AND SEROLOGIC METHODS
 Binding of antigen to a specific antibody
S (Sample well) – Where you put the sample
 Serum is the sample; Needs particular amount of drops HISTOPATHOLOGY
II (Two lines) – Reactive  Combination of histology and pathology
Cl (Control line) – Non-reactive; Test is valid  Also called “Anatomic Pathology”
TI (Test line) – Invalidate the test because C has no line  The art and science of producing a quality tissue section to enable the
pathologist to diagnose the presence or absence of a disease
 Should be timed (timer is needed); read immediately after the  Autopsy and biopsy specimens
time PIONEERS IN HISTOPATHOLOGY
 If checked late – The line will fade  Ferdinand Blum
 Check with co-MT for the result  Proposed the use of formaldehyde as a fixative
 LIGHT LINE – virus is not that abundant TESTS IN ANATOMIC PATHOLOGY
 Dengue Duo Test  Routine histopathologic examination
 A rapid test designed to detect NS1 Antigen (Non-  Fine needle aspiration biopsy
Structural Protein 1); If there is = Positive  Papanicolaou smear
 Antigen released by the mosquito  Cytopathological techniques:
 Not solely in dengue  Cell block, cytospin
 Antibodies (IgG and IgM)  Frozen section
 IgM – infection is new  Breast panel
 Both – Infection has been there for a long  Special staining
time; Had dengue before and it returned  Immunohistochemical staining
(Dengue has 4 strains)  Post-mortem examination
 IgG – Old infection or a different disease ROUTINE EXAMINATION
 Both tests are expensive NUMBERING
 Invalid if there is no line in C  Recording the tissue specimen in a logbook and assigning identification
 If the result is positive = Kit is kept for evidence (Include numbers to the received specimen
patient details)
 If the result is negative = Throw kit away because it is
infectious

GROSS EXAMINATION
FIXATION
 Preserving the tissue specimen in as life-like a manner as possible
 Systematic Lupus Erythematosus (SLE) Latex Test
 Routine fixative: 10% formalin
 Detects presence of antinuclear antibodies associated
with SLE
 Rheumatoid Factor (RF) Latex Test
 Used to determine rheumatoid arthritis
 Reagent contains latex which causes agglutination
 Human Leukocyte Antigen (HLA)
 To determine compatibility in organ, tissue, and bone
marrow transplantation DECALCIFICATION
 To determine paternity (NEW NAME: DNA Testing)  Removal of calcium from some tissues or organs
 To diagnose HLA-related disorders such as autoimmune  Routine decalcifying agent: Nitric Acid
diseases
 Not found in Davao, only in Manila; Very expensive
MACHINES IN THE IMMUNOLOGY-SEROLOGY AREA
 Mini-VIDAS
 Place the sample in the cartridge and push buttons and
the result will be seen
 Can be put on the table top

DEHYDRATION
 Removing water from the specimen by using the increasing grades of
ethyl alcohol

 Architect i1000SR
 Fun automation machine
 Bigger
 Has a monitor and a connected computer that transmits
the results
 Reagents can be found inside (Also tubes)

CLEARING
 Removing excess alcohol in tissues
 Makes tissues transparent
  Routine cleaning agent: Xylene
INFILTRATION
 Filling up tissue spaces or cavities with melted paraffin wax
EMBEDDING
 Placing the infiltrated tissue inside a mold

TRIMMING
 Removing excess paraffin wax from the block

CYTOSPIN
 To concentrate cells on a slide in a uniform monolayer using a high-
speed centrifuge

SECTIONING
 Employs the use of hematoxylin and cosin dyes to differentiate the
cells and the cell constituents
MOUNTING
 Putting the cover slip on the stained tissues using a mounting medium

FROZEN SECTION
 Performed when an immediate or rapid microscopic analysis of
specimen is needed
LABELLING  Cryostat
 Specimen number is indicated on the glass slide
FINE NEEDLE ASPIRATION BIOPSY
 To investigate superficial masses or lumps to detect any pathologic
conditions like malignancy
 Insertion of a hollow needle into the mass for sample collection

BREAST PANEL
 Biomarkers important in the genetic testing for breast cancer
 Estrogen receptor
 Progesterone receptor
 HER2-NFU
 P-53
 DNA Ploidy analysis
HISTOCHEMISTRY
 Uses special stains to determine the chemical compounds and their
distribution within and in between the biological cells of the body
IMMUNOHISTOCHEMICAL STAINING
PAPANICOLAOU SMEAR/PAP SMEAR  Detecting antigens in the cells of tissue sections by using antibodies
 Screening for cervical cancer and any pre-cancerous changes in the
cervix
 Also done to detect STD’s such as trichomonas, candidiasis, and human
papillomavirus (HPV)

POST MORTEM EXAMINATION/AUTOPSY

 Thorough examination of a dead body to determine the cause of death,
manner of death, and to evaluate any disease or injury that may have
been present

 Section in the laboratory

 CBC is part of the annual physical laboratory exam together with
CELL BLOCK fecalysis, urinalysis, x-ray
 Paraffin-embedded specimen prepared from dried mucus, sputum,  The science of studying the blood and its components
and debris found in body fluids  Comes from the Greek words: “Haima” – blood & “Logos” – science
of
PHLEBOTOMY
 Standard procedure of blood collection
 Must be learned
 TYPES:
 Skin Puncture – Capillaries using a lancet (has lots of types)
 Punctures skin in fingers (Middle or ring finger)
 Autolancet – Covered in plastic and has a spring inside
  Color coded in order to trick children or for children to play
with
 Needle is not exposed

 FUNCTIONS:
SYRINGE  Transports RBC, WBC, and platelets through the blood
 Venipuncture = veins using: vessels
 The 3 most common methods of blood collection using  Remove waste products of metabolism
venipuncture  Blood will pass through and be filtered in
1. Syringe kidneys
2. Butterfly Infusion Set – Used for gluta and  Waste product is in the form of urine
other medicine RED BLOOD CELL (ERYTHROCYTE)
3. Evacuated Tube System (ETS) – Uses 2 way  Anucleated biconcave cells produced from the bone marrow
needle  1st formed element found in the bottom
 Adapter (Plastic covering)  Hemoglobin = gas transporting protein molecule; Major component of
 2-way needle – cannot be
RBC
determined if blood is hit
 Hem = Nitrogenous substance (Contains irons in the ferrous
 Arterial Puncture – Arteries done by respiratory therapists
state)
 Much more dangerous because it looks for the pulse; Quite  Globin = Protein portion
painful
 Functions:
 45 angle (needle) so that it won’t hurt so much
 Transport oxygen (Lungs → Tissues); Inspiration = Inhale
 TEST: Arterial Blood Gas (ABG) is determined oxygen
 Transport carbon dioxide (Tissue → Lungs); Expiration =
Exhale carbon dioxide
 Mature RBC does not have a nucleus; Only the younger ones
 Small parts are the platelets
 In the cell there is a CENTRAL PALLOR – cavity

AUTOLANCET

EVACUATED TUBE SYSTEM (ETS) ELECTRON MICROSCOPIC PICTURE OF A RED BLOOD CELL
 Lavender Top – Most common tube used
 Contains the anticoagulant: EDTA or Ethylenediamine
Tetraacetic Acid
 RBC is found at the bottom (Heavy substance)
 Blood is not centrifuged

BLOOD FILM (SMEAR) MICROSCOPIC PIC

 2 types of electron microscopic used to view RBC
 Transmission Electron Microscope (TEM)
 Scanning Electron Microscope (SEM)
 Associated conditions:
 Anemia – Low number of RBC’s (low RBC count); Not a
 Blood is centrifuged disease
 Polycythemia – High number of RBC’s (high RBC count)
 Average total volume of blood = 5L (liters)

PLASMA
 Clear, yellow liquid
 Water, sugar, fat, protein, and salt solution
 Composed majorly of water – 95% and the rest are in fraction
components (Include other vitamins and minerals)
 Blood is a nutritive material

WHITE BLOOD CELL (LEUKOCYTE)

 Nucleated cells that lack hemoglobin (Has a nucleus compared to RBC)
  Acts as defense against infections
 Leukocytosis – Increased number of leukocyte; Osis –
increase
 Leukopenia – Decreased number of leukocytes; Penia –
Decrease

MONOCYTE
 Biggest among the 5 WBCs
 Kidney-shaped or horseshoe-shaped nucleus
 Has an indention or indentation
 Has a large cytoplasm; Large cytoplasm = Large cell
 Monocyte is what is called during circulation in the blood vessels;
Outside there are 2 types (In tissues)
 Dendritic Cell – Marks out cells that are antigens (foreign
bodies) that should be destroyed by lymphocytes
NEUTROPHIL  Macrophage – Act as antigen-presenting cells; Has
 Most numerous different names based on where they reside in the body
 Most phagocytic  E.g. Cockfur cells (liver); splenic macrophage
(spleen)
 Primary response to an infection
 Multi-lobed nucleus – Has 3-5 lobes (multinucleated); If there are more
lobes, there is an underlying condition (usually 8 lobes)
 Pale lilac granules
 Has fine granules (small) – Granulated
 Function: Immune defense (Phagocytosis)
PLATELETS (THROMBOCYTES)
 Color lavender
 Cell fragments; Not cells but cell fragments which come from a big
cell: MEGAKARYOCYTES – Comes from the bone marrow; 10-15x
larger than normal RBC (50-100 m in diameter)
 When it sheds off, the shedding are known as the platelets
LYMPHOCYTE  Function:
 Spherical nucleus  Form clots during injury to prevent blood from leaking out
 “Robin’s Egg Blue” Cytoplasm  Acute blood loss can kill a person
 Nucleus occupies entirely the cell (less cytoplasm) Platelet plug formed by platelets → Forms clot (Fibrin clot) → Hardens the wound
 Types:  Thrombocytosis – Increased number of platelets
 T-cell – Cellular immune response  Thrombocytopenia – Decreased number of platelets
 B-cell – Antibody production
 NK Cells (Natural Killer Cells) – Kills cancer cells; Combats
cancer cells
 Cancer – Easily metastasizes (Metastasis);
Most common cancer is women – Breast
cancer

EOSINOPHIL HEMATOLOGY SECTION

 Usually with a bilobed nucleus (But can have up to 3 lobes)  Sample: Whole blood and blood film (smear)
 Granules sarin bright-reddish orange (Bright color is the indicator) Complete Blood Count (CBC)
 Has large granules  Not required all at once (separately)
 Slightly larger than neutrophil  Hemoglobin
 Late to arrive because of its bigger size, it has a sluggish movement  Hematocrit
compared to neutrophil  Red Blood Cell count
 Functions:  White Blood Cell count
 Defense against parasites (Arises during parasitism)  Platelet count (estimate)
 Activates allergic response  RBC indices – Checks morphology of anemia; Classification
and quality control (QC)
 Mean Cell Volume (MCV) – Refers to the size
of RBC
 Mean Cell Hemoglobin (MCH) – Refers to the
hemoglobin color
 Mean Cell Hemoglobin Content (MCHC) – Refers
to the hemoglobin color
BASOPHIL
 Most common type of anemia = Iron deficiency anemia – Indices are
 3rd member of granulocytes
low; RBC is microcytic (RBC is small in size) and hypochromic (lacks
 With purple-blue/blue-black granules
hemoglobin content)
 Obscures the cell (Nucleus cannot be seen) PROCEDURES PERFORMED IN THE HEMATOLOGY SECTION
 Usually bilobed 1. Counting the number or concentration of cells
 Functions: 2. Determining the relative distribution of various types of cells
 Inflammatory response 3. Measuring biochemical abnormalities of blood
 Involved in allergic response 4. Homeostasis and coagulation assays (PT- Prothrombin time; APTT-
Activated Partial Thromboplastin Time
TESTS PERFORMED IN HEMATOLOGY SECTION
HEMOGLOBIN DETERMINATION
 Hemoglobin – Iron-containing oxygen transport metalloprotein in the
red blood cells
  Metal – Iron
 Methods:
 Cyanmethemoglobin Method
 Reference method
 Reagent: Drabkin’s Reagent
 Principle: Oxidation of ferrous iron (2+) to
ferric iron (3+) by potassium ferricyanide
(reagent) = Methemolgobin
 Methemoglobin is converted to
cyanmethemoglobin by virtue of potassium
cyanide ions
 Drabkin’s reagent is no longer used because of
presence of cyanide (Poses a threat to health
because of its toxicity; Cyanide poisoning)
 Now: Modified Drabkin’s Reagent
(No presence of cyanide)
 Color of reagent = Yellow
 Automated Hemoglobinometry
 Utilizes cyanmethemoglobin method with
modified Drabkin’s reagent – No cyanide
 Point-of-Care (POC) Hemoglobin Assay
 Brought to the patients in their bedside
 HemoCue method
 Modified azidemethemoglobin reaction
 Reactants: sodium deoxycholate, sodium nitrite,
and sodium azide
Thoma Pipette – Most important; Has blood and diluting fluid
Manual diluting fluid used for WBC
 2% glacial acetic acid
 1% hyaluronic acid
Function of diluting fluid – Hides RBC so that WBC will be left and is easier to count
Hemacytometer with cover slip – Slide which is quite thick; Also important
BLOOD CELL COUNT – AUTOMATED
1. Electrical Impedance
 Also known as Coulter Principle
 Sizing and counting of particles is based on changes in
electrical resistance creating voltage pulses
 Pulse will become number which will become the result
2. Optical Detection
 Hydrodynamic focusing method
 Uses laser light in cell counting and sizing
 A combination of principles in cell counting
PERIPHERAL BLOOD SMEAR

HEMATOCRIT DETERMINATION
 Also known as Packed Cell Volume (PCV) or Erythrocyte Volume
Fraction (EVF)
 For evaluation or treatment of anemia and determine presence of
nutritional deficiencies
 Methods – Done in the laboratory
 Unit of Measurement:
 Hemoglobin – g/dL or g/L
 Hematocrit - % or L/L (SI unit)
A. Spun Microhematocrit
 Manual
 Blood collection method = Skin puncture
 Spin a blood-filled capillary tube using a microhematocrit
centrifuge (Spin for 3-5 minutes)
 Read in Hematocrit reader
 Capillary Tube Sealing Clay – To seal the capillary tube so
blood won’t spill
B. Automated
 Computed from the mean cell volume and the red cell count

HOW TO MAKE A GOOD SMEAR

1. Needs 2 slides: Spreader slide and glass slide
2. Drop blood and use spreader slide to create smear

 It is important to have a good transition from thick to thin until the

feathered edge (Create a thumb shape)
 Place name of patient by the frosted slide using pencil (So that it won’t
be erased by the alcohol)
 Check the distribution of the cell (RBC’s barely touch one another);
Identification of cells
BLOOD CELL COUNT – MANUAL ADDITIONAL HEMATOLOGY PROCEDURES
RETICULOCYTE COUNT – Done in Hematology 2
 Reticulocytes: Young RBC’s without nucleus but still bears cytoplasmic
RNA
 RBC has 6 stages of development (Reticulocyte is in the 5th stage)
 Determines how the bone marrow produce and release new RBC’s to
compensate lost/damaged RBCs
 Blood film is stained with supravital stain
 Reticulum: Remnants of RNA

ERYTHROCYTE SEDIMENTATION RATE – Done in Hematology 2 Laboratory
 Rate at which RBCs fall in a column in known as erythrocyte
sedimentation rate
 Non-specific test for inflammation
 Reference method: Westergren Method
 Anticoagulant = Sodium citrate
 Time of testing = One hour
 Sample: Whole blood
 Unit of measurement: mm/h – Millimeters per hour
 For Reticulum - % (Unit of measurement)
 Result is seen by virtue of gravity (RBC goes down)
PLASMA COAGULATION ASSAYS
PROTHROMBIN TIME (PT) and ACTIVATED PARTIAL THROMBOPLASTIN TIME (APTT)
 Coagulation testing
 Detects abnormalities in hemostasis (Coagulation)
 Anticoagulant: Sodium citrate
 Sample: Plasma only
 After centrifugation, get the plasma
 Centrifugation time: 15 minutes (Longer centrifugation time)
 A branch of immunology which deals with the uses of immunologic
principles to study and identify the different blood groups
 It reflects the importance of the blood bank and the MT profession
in ensuring the safety and welfare of patients that require blood
transfusion
BLOOD BANK
 A separate area in a clinical laboratory hospital where blood is collected
from donors
 Performs ABO and Rh typing
 Prepares blood and blood components for transfusion
 Blood that is transfused into a recipient must be tested first to ensure
compatibility with the recipient’s blood
 To reduce the risk of transfusion reactions
 To ensure that the blood/blood components are safe
ABO BLOOD GROUP SYSTEM
 Discovered by Karl Landsteiner (1900’s) and received the Nobel Prize
(1930)
 He categorized the blood groups as A, B, and O
 Based on the presence of agglutinating antibodies in the
serum/plasma of individuals who do not possess the
corresponding ABO antigen
 AB – 4th major ABO Blood type discovered by Alfred von Decastello
and Adriano Sturli
ABO BLOOD TYPING
 A test to determine the blood type of an individual
 Cell typing (Direct or Forward typing)
 To determine antigens in the RBCs of an
individual by using commercially prepared
antisera of known specificity
 Serum typing (Backward, direct or indirect typing)
 To determine antibodies in the serum/plasma
of an individual by using RBCs of known
specificity
Rh BLOOD GROUP SYSTEM
 Discovered by Karl Landsteiner and Alexander Weiner (1940)
 They injected rabbits with Rhesus Macaque monkey RBCs
and Rh antibodies were produced
 Rh antibodies + human RBC’s = AGGLUTINATION = Rh Positive
 Rh antibodies + human RBC’s = NO AGGLUTINATION = Rh Negative
 5 important Rh antigens = D,C,F,e,c
 D antigen = Most important and immunogenic antigen
  Rh typing – Based on the presence and absence of the D antigen on 8. PLATELET CONCENTRATE
the surface of RBC’s using commercially prepared anti-D antisera  Indications: Thrombocytopenia & Platelet dysfunction

9. CRYOPRECIPITATE
 Indications: Fibrinogen deficiency

COMPATIBILITY TEST
 It is a series of procedures assigned to ensure the safety of
transferring blood
 It must be performed before the transfusion of blood
components
 Blood typing and crossmatching must be done to prevent
harmful transfusion reactions between the recipient
blood and the donor blood
 2 Parts:
 Major Crossmatching 8.
 Patient serum is mixed with the donor RBCs
 Detects if there are antibodies in the patient
serum that can destroy the transfused RBC
from the donor
 RS-DR
 Minor Crossmatching
 Patient RBCs are mixed with the donor RBCs
 Detects if there are antibodies in the donor
serum that can destroy the patients RBCs
 PR-DS
BLOOD COMPONENTS and their INDICATORS
1. WHOLE BLOOD
 Effect: Volume replacement and restoration of oxygen-
carrying capacity
 Indications: Acute blood loss
 Irradiated Whole Blood: Avoidance TA-GVHD

2. PACKED RED BLOOD CELL (PRBC)

 Effect: Restoring oxygen carrying capacity
 Indication: Anemic conditions with hypoxia

3. WASHED PRBC
 Indications: Allergic response to plasma proteins
4. LEUKOCYTE-REDUCED PRBC
 Indications: Febrile transfusion reactions

5. FROZEN PRBC
 Indications: Unusual blood types
6. IRRADIATED PRBC
 Indications: Avoidance TA-GVHD
7. FRESH FROZEN PLASMA (FFP)
 Effect: Replacement of plasma factors
 Indicators: Severe bleeding in unknown factor deficiency