MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

SAFETY STANDARDS AND AGENCIES

 OSHA Occupational Safety and Health
Administration
 CLSI Clinical and Laboratory Standards Institute
 CDC Centers for Disease Control and Prevention
 CAP College of American Pathologists
 TJC The Joint Commission (Formerly Jcaho)

 OSHA: Occupational Health and Safety

Administration within the U.S. Department of
Labor to set levels of safety and health for all
workers in the United States.
 CLSI: Clinical and Laboratory Standards
Institute Nonprofits educational organization
that sets voluntary consensus standards for all
areas of clinical laboratories;
 CDC: Center for Disease Control and Prevention
Federal agency that carries out mandated public
health laws and reporting requirements.
TYPES OF SAFETY HAZARD
ALL CLINICAL LAB SHOULD HAVE: TYPE SOURCE POSSIBLE
INJURY
 Chemical Hygiene plan
Biological Infectious (infections)
 Exposure control plan agents Bacterial, fungal,
 Copy of MSDS viral, or parasitic
Sharps Needles, Cuts, punctures or
SAFETY BEGINS WITH THE RECOGNITION OF lancets, blood-born
HAZARDS AND IS ACHIEVED THROUGH THE FF: broken glass pathogen
 Application of common sense exposure
 Listen to the instructions Chemical Preservatives Exposure to toxic,
 A safety-focused attitude and reagents carcinogenic, or
 Good personal behavior caustic agents
Radioactive Equipment Radiation
 Good housekeeping in all laboratory work and
and exposure
storage areas
radioisotopes
 Continual practice of good laboratory technique. Electrical Ungrounded Burns or shock
TWO PRIMARY CAUSES OF ACCIDENTS: or wet
 Unsafe acts equipment;
 Unsafe environmental conditions frayed cords
SAFETY EQUIPMENT Fire/ Bunsen Burns,
 Safety showers and Eyewash stations explosive B burners, dismemberment
 Fire extinguishers organic
 Fume Hoods chemicals
Physical Wet floors, Falls, sprains or
 Biosafety Cabinets
heavy boxes, strains
 Complete PPE patients
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 Category III – no exposure to blood and body
fluids
 These microorganisms are frequently present in  Employers must offer HBV vaccine to all personnel
the specimens received in the clinical (Category I and II)
laboratory.
 CDC  Universal Precautions (1987)
 Blood and body fluid precautions should be
consistently used for all patients
 Specimens should be “capped” during
centrifugation

 *Any blood, body fluid, or other potentially

infectious material spill must be cleaned up
using:
 Spill cleanup kit
 Common aqueous detergent
 10% bleach using appropriate contact time.

STEPS TO CLEAN BLOOD SPILL

 Wear PPE
 use forceps for discarding the broken glass on a
sharps container
 Cover with absorbent paper towels
 Flood area with 10% bleach solution
 Let sit for 10 min.
 Clean up area with paper towels
 Dispose in Biohazard bag
 Repeat if necessary.
OSHA BLOOD-BORNE PATHOGENS STANDARD REQUIRES
WRITTEN “EXPOSURE CONTROL PLAN”
Categories of Exposure:
 Category I – daily exposure to blood and body
fluids
 Category II – regular exposure to blood and body
fluids
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 Fire and explosion hazards
 Reactivity data
 Spill and disposal procedures
 Chemical spills: When skin contact occurs, the  PPE recommendations
best first aid is to flush the area with large  Handling
amounts of water for at least and then seek  Emergency and first aid procedures
medical attention.  Storage and transportation precautions
 Chemical Handling: Chemicals should never be  Chemical manufacturer’s name, address, and
mixed together unless specific instructions are telephone number
followed, and they must be added in the order  Special information section
specified.
 This is particularly important when
combining acid and water.
 Chemical Hygiene Plan: OSHA also requires all
facilities that use hazardous chemicals to have a  The National Fire Protec=on Association (NFPA)
written chemical hygiene plan (CHP) available has developed the Standard System for
to employees. The purpose of the plan is to detail providing codes and standard information about
the following: the chemicals/solutions.
 Appropriate work practices, Standard
operating procedures, PPE, Engineering
controls, such as fume hoods and
flammable safety cabinets, Employee
training equipment's and Medical
consultation guidelines.
STORAGE AND HANDLING OF
CHEMICALS
FLAMMABLE/COMBUSTIBLE CHEMICALS
 Classified according to flash point  the
temperature at which sufficient vapor is given
off to form an ignitable mixture with air
CORROSIVE CHEMICALS
 injurious to the skin or eyes by direct contact or
to the tissue of the respiratory and
gastrointestinal tracts if inhaled or ingested
REACTIVE CHEMICALS
 spontaneously explode or ignite or that evolve
heat or flammable or explosive gases
CARCINOGENIC CHEMICALS

MSDS
 Product name and identification
 Hazardous ingredients
 Permissible exposure limit (PEL)
 Physical and chemical data
 Health hazard data and carcinogenic potential
 Primary routes of entry

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
FIRE/ EXPLOSIVE HAZARDS RADIATION HAZARD
 The Joint Commission on Accreditation of  Equipment and radioisotopes
Healthcare Organizations (JCAHO) requires that  Radiation Safety
all health-care institutions post evacuation - All areas where radioactive materials are
routes and detailed plans to follow in the event used or stored must be posted with caution
of a fire. signs, and traffic in these areas should be
 Laboratory personnel should be familiar with restricted to essential personnel only.
these procedures. When a fire is discovered, all - Radiation monitoring utilizes film badge or
employees are expected to take the actions in survey meter
the acronym:  maximum permissible dose is 5000 mrem/year
 Rescue whole body.
 Alarm MECHANICAL HAZARDS
 Contain  Centrifuges  must be balanced to distribute
 Extinguish the load equally.
FIRE HOTLINE:  Never open the lid until the rotor has
 ANTIPOLO CENTRAL FIRE STATION (BUREAU come to a complete stop
OF FIRE PROTECTION OFFICE)  871-2865  Safety locks on equipment should never
 MAYAMOT SUB-STATION  250-0497 be rendered inoperable
ELECTRICAL HAZARD  Glass beads – help eliminate bumping/boil over
 Ungrounded or when liquids are heated
wet equipment;  Infectious sharps - disposed in OSHA-approved
frayed cords containers.
 Electrical
Safety
 Lock-out or tag  4 Basic Waste Disposal Technique
malfunctioning  Flushing down the drain
electrical or  Incineration
mechanical  Landfill burial
equipment  Recycling
until serviced CHEMICAL WASTE
 Know how to  Flush water-soluble substances down the drain
knock a shocked person loose using a non- with large quantities of water
conductive material.  Strong acids and bases should be neutralized
Electrical Precautionary Procedures: before disposal
 Use only explosion-proof equipment in  Foul smelling chemicals should never be
hazardous atmospheres. disposed down the drain
 Be particularly careful when operating high  Flammable solvents  collected in approved
voltage equipment, such as electrophoresis containers
apparatus.  Flammable material specially designed
 Check for frayed electrical cords. incinerators
 Promptly report any malfunctions or equipment  Solid chemicals  landfill
 Do not work on “live” electrical equipment. RADIOACTIVE WASTE
 Never operate electrical equipment with wet  depends on the type of waste (soluble or non-
hands. soluble), its level of radioactivity, and the
 Know the exact location of the electrical control radiotoxicity and half-life of the isotopes
panel for the electricity to your work area. involved.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
BIOHAZARDOUS WASTE
 All biological waste (EXCEPT URINE) should be
placed in appropriate containers labeled with
biohazard symbol.
 URINE: may be discarded by pouring it into the
lab sink.
 The sink should be flashed also with
water after the urine has been discarded.
 Decontaminate the sink by 1:5 or 1:10
dilution of sodium hypochlorite (bleach
solution).
 Incineration, inactivation, burial, chemical
disinfection, encapsulayion in a solid matrix.
Inactivation:
 Heat sterilization (250oC for 15 minutes)
 Ethylene Oxide (450-500 mg/L at 55-60oC )
 2% Glutaraldehyde
 10% hydrogen peroxide
 5.25 hypochlorite (bleach)
 10% (v/v with tap water) of common household PROPER HAND WASHING
bleach)  HBV (10 minutes), HIV (2 minutes)  After completing lab work, and before leaving
the laboratory.
 After removing gloves.
 Before eating, drinking, applying makeup, and
changing contact lenses, and before and after
using the lavatory.
 Before all activities involving hand contact with
mucous membranes or skin breaks.
 Immediately after accidental skin contact with
blood, body fluids, or issues.

 Wet hands with warm water.

 Apply antimicrobial soap.
 Rub to form a lather, create friction, and loosen
debris.
 Thoroughly clean between fingers, including
thumbs, under fingernails and rings, and up to
the wrist, for at least 20 seconds.
 Rinse hands in a downward position.
 Dry with a paper towel.
 Turn off faucets with a clean paper towel to
prevent recontamination.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

PHLEBOTOMY
 Defined as an incision or a puncture into a vein
in order to obtain blood
 One of the oldest medical procedures, dating Bloodletting-Set of a Barber-Surgeon
back to the early Egyptians and was termed as
“bloodletting” Barbers’ Poles

 Hippocrates believed that disease was caused

by an excess of body fluids, including blood, bile,
and phlegm, and that removal of the excess
would cause the body to return to or maintain a
healthy state, and the first-line treatment before
all others: bloodletting.
 Techniques for bloodletting:
 -suction cup devices with lancets
 -application of blood-sucking worms,
called “leeches,”
 -barber surgery, in which blood from an
incision produced by the barber’s razor
was collected in a bleeding bowl.
 Bloodletting is now called “therapeutic
phlebotomy” and is used as a treatment for only
a small number of blood disorders such as
hemochromatosis.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

BLOODLETTING THROUGH SUCTION DEVICES

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
PHLEBOTOMY AT PRESENT
 primary role of phlebotomy is the collection of
blood samples for laboratory analysis to
diagnose and monitor medical conditions.
 Because of the increased number and
complexity of laboratory tests, phlebotomy has
become a specialized area of clinical laboratory
practice and has brought about the creation of
the job title “phlebotomist.”
DUTIES OF THE PHLEBOTOMIST
IN TODAY’S HEALTHCARE
SETTING
 A phlebotomist is a person trained to obtain
blood samples primarily by venipuncture and
microtechniques.
 Major traditional duties and responsibilities of
the phlebotomist include:
 Correct identification and preparation of
the patient before sample collection
 Collection of the appropriate amount of
blood by venipuncture or dermal puncture
for the specified tests
 Selection of the appropriate sample
containers for the specified tests
 Correct labeling of all samples with the
required information
 Appropriate transportation of samples
back to the laboratory in a timely manner
 Effective interaction with patients and
hospital personnel
 Processing of samples for delivery to the
appropriate laboratory departments
 Performance of computer operations and
record-keeping pertaining to phlebotomy
 Observation of all safety regulations,
quality control checks, and preventive
maintenance procedures
 Attendance at continuing education
programs
 Monitoring the quality of samples collected
on the units
 Performing and monitoring point-of-care
testing (POCT).
Traits That Form the Professional Image of the
Phlebotomist
 Dependable, cooperative, committed
 Compassionate, courteous, respectful
 Integrity, honesty, competence
 Organized, responsible, flexible
 Appearance
 Communication
Dependable, Cooperative, Committed
 Laboratory testing begins with sample
collection and relies on the phlebotomist to
report to work whenever scheduled and on
time.. Failure to appear or arriving late puts
additional pressure on the staff members
present.
 Be willing to demonstrate your commitment to
your job and your cooperation to assist fellow
employees. A committed phlebotomist attends
staff meetings, reads pertinent memoranda, and
observes notices placed on bulletin boards or in
newsletters.
Compassionate, Courteous, Respectful
 Phlebotomists deal with sick, anxious, and
frightened patients every day. They must be
sensitive to their needs, understand a patient’s
concern about a possible diagnosis or just the
fear of a needle, and take the time to reassure
each patient. A smile and a cheerful tone of voice
are simple techniques that can put a patient
more at ease.
 Courteous phlebotomists introduce themselves
to the patients before they approach them. This
also aids in identifying the patient as you can
then ask them to state their name in the same
conversation. Phlebotomists must also
understand and respect the cultural diversity of
their patients.
 Cultural diversity includes not only language
but also religious beliefs, customs, and values.
Do not expect every patient to respond to you in
the same way and do not force your mannerisms
and approach on them.
Honesty, Integrity, Competence
 The phlebotomist should never hesitate to admit
a mistake, because a misidentified patient or
mislabeled sample can be critical to patient
safety. Patient confidentiality must be protected,
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
and patient information is never discussed with
anyone who does not have a professional need
to know it.
 Phlebotomists must demonstrate competence in
the procedures they are trained to perform.
However, overconfidence in one’s abilities can
result in serious errors. Never perform a
procedure that you have not been trained to
perform. When faced with this situation do not
hesitate to ask for assistance from someone
more experienced.
Organized, Responsible, Flexible
 Phlebotomists need to organize their collection
equipment and maintain well-stocked collection
tray or station. They must also organize and
prioritize their work.
Appearance
 Phlebotomists should be neat and should have
clean-looking appearance that portrays a
professional attitude to the patient. Remember
first impressions are lasting impressions often
made within 30 seconds and the phlebotomist
represents the entire laboratory staff.
 Clothing and lab coats must be clean and
unwrinkled. Clothing worn under the laboratory
coat should meet institutional requirements.
Lab coats must be completely buttoned and
completely cover clothing.
 Shoes must be clean, polished, closed toed, and
skid-proof.
 If jewelry is worn, it must be conservative.
Dangling jewelry including earrings can be
grabbed by a patient or become tangled in
bedside equipment. Many institutions do not
permit facial piercings and tattoos; if present,
they must be completely covered. Makeup must
also be conservatively applied.
 Perfume and cologne are usually not
recommended or must be kept to a minimum.
Many persons are allergic to certain fragrances.
Remember the phlebotomist works in close
contact with the patient and the smell of
perfume can be particularly disturbing to a sick
person.
 Hair including facial hair must be clean, neat,
and trimmed. Long hair must be neatly pulled
back.
 Personal hygiene is extremely important
because of close patient contact, and careful
attention should be paid to bathing and the use
of deodorants and mouthwashes.
 Fingernails must be clean and short. Based on
the Centers for Disease Control and Prevention
(CDC) Handwashing Guidelines, artificial nail
extenders are not allowed.
Communication Skills
 Good communication skills are needed for the
phlebotomist to function as the liaison between
the laboratory and the patients, their family and
visitors, and other health-care personnel.
 The three components of communication:
 verbal skills
 listening skills
 nonverbal skills or body language
VERBAL SKILLS
 to introduce themselves, explain the procedure,
reassure the patient, and help assure the patient
that the procedure is being competently
performed.
 Barriers to verbal communication that must be
considered include physical handicaps such as
hearing impairment; patient emotions; and the
level of patient education, age, and language
proficiency.
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
LISTENING SKILLS
 Looking directly and attentively at the patient
 Encouraging the patient to express feelings,
anxieties, and concerns
 Allowing the patient time to describe why he or
she is concerned
 Providing feedback to the patient through
appropriate responses
 Encouraging patient communication by asking
questions
NONVERBAL SKILLS
 include facial expressions, posture, and eye
contact.
 If you walk briskly into the room, smile, and look
directly at the patient while talking, you
demonstrate positive body language. This
makes patients feel that they are important and
that you care about them and your work
 Allowing patients to maintain their zone of
comfort (space) is important in phlebotomy
even though you must be close to them to collect
the sample.
TELEPHONE SKILLS
 The phlebotomy department frequently acts as
a type of switchboard for the rest of the
laboratory because of its location in the central
processing area.
 This is a prime example of the phlebotomist’s
role as a liaison for the laboratory, and poor
telephone skills affect the image of the
laboratory.
To observe the rules of proper telephone etiquette:
 Answer the phone promptly and politely, stating
the name of the department and your name.
 Always check for an emergency before putting
someone on hold, and return to calls that are on
hold as soon as possible.
 Keep writing materials beside the phone to
record information such as the location of
emergency blood collections, requests for test
results, and numbers for returning calls.
 Make every attempt to help callers, and if you
cannot help them, transfer them to another
person or department that can.
 Provide accurate and consistent information by
keeping current with laboratory policies,
looking up information published in department
manuals, or asking a supervisor.
 Speak clearly and make sure you understand
what the caller is asking and that he or she
understands the information you are providing.
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
WHOLE BLOOD
 Has Plasma and formed elements (unclotted).
BLOOD
 Liquid portion of clotted blood
 Without anticoagulant
 Contains albumin and globulin
 Liquid portion of unclotted blood
 With anticoagulant
 Contains albumin, globulin and fibrinogen
DIET
 The tests most affected are glucose and
triglycerides.
 Serum or plasma collected from patients shortly
after a meal may appear cloudy or turbid
(lipemic) due to the presence of fatty
compounds such as meat, cheese, butter, and
cream.
 -Alcohol consumption  transient elevation in
glucose and  chronic consumption  liver
function tests and triglycerides
 -Caffeine  hormone levels
POSTURE
 Can cause variations in some blood constituents,
such as cellular elements, plasma proteins,
compounds bound to plasma proteins, and high
molecular weight substances.
EXERCISE
 Increased activity of muscle enzymes
 Elevated concentration of sex hormones
 Elevated concentration of steroids
STRESS
 Nervous patient before sample collection may
increase levels of adrenal hormones, increase
WBC counts, decrease serum iron, and markedly
affect arterial blood gas (ABG) results.
SMOKING
 Acute effects: increase in glucose, BUN,
cholesterol and triglycerides
 -Chronic effects: Increase in blood hemoglobin
values (carboxyhemoglobin)
 Decrease in IgG, IgA, and IgM  weak immune
system
ALTITUDE
 RBC counts and hemoglobin (Hgb) and
hematocrit (Hct) levels are increased in high-
altitude areas such as the mountains where
there are reduced oxygen levels.
AGE AND GENDER
 Laboratory results vary between infancy,
childhood, adulthood, and the elderly  gradual
change in the composition of body fluids.
 Hormone levels vary with age and gender
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 RBC, Hgb, and Hct values  higher in male
patients
 Needle size varies by both length and gauge
PREGNANCY (diameter)
 Needle gauge: refers to the diameter of the
 caused by the physiological changes in the
needle bore.
body including increases in plasma volume.
 The smaller the gauge number the bigger the
ALCOHOL INGESTION diameter of the needle.
 Needles should be visually examined before use
to determine if any structural defects, such as
nonbeveled points or bent shafts, are
present.
 Venipuncture- as the name implies, it is the
puncture to a vein to for collecting blood
sample.
 The first step in learning to perform a
venipuncture is knowledge of the needed
equipment.

EVACUATED TUBE SYSTEM

 Blood is collected directly into the evacuated
tube, eliminating the need for transfer of
specimens and minimizing the risk of
biohazard exposure

 Double-Pointed Needle
 Needle Holder
 Color-coded Evacuated tubes

 POINT: Sharpened end of the needle

 Bevel: The end of the shaft that forms a flat,
slanted surface
 Lumen/ BORE: the hollow core of the
needle/opening
 Shaft: the long slender stem of the needle
 HUB: where the needle is attached
 Rubber sleeve: for 2 way needle. Where the
tube is punctured
 Needle Size for Venipuncture: 1-inch and
1.5-inch lengths are used.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 To protect phlebotomists from accidental

needlesticks by contaminated needles

 Rigid, puncture-resistant, leak-proof

disposable “sharps” containers labeled
BIOHAZARD that are easily sealed and
locked when full.

Eclipse blood collection needle (Becton,

Dickinson, Franklin Lakes, NJ) uses a shield that the
phlebotomist locks over the needle tip after
completion of the venipuncture.

 made of rigid plastic and may be designed to

act as a safety shield for the used needle.
 Occupational Safety and Health
Administration (OSHA) directs that holders COLLECTION TUBES
must be discarded with the used needle.

EVACUATED TUBES
 also known as Vacutainers and are available in
glass and plastic.
 Contain a premeasured amount of vacuum for
blood collection
 The amount of blood collected in an evacuated
tube ranges from 1.8 to 15 mL and is
determined by the size of the tube and the
amount of vacuum present.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 EDTA: 8x
 8X EDTA, PINK, GREEN, LIGHT GREEN,  Pink:8x
ORANGE, GRAY, TAN, ROYAL BLUE,  Light blue: 3-4x
 Gold: 5x blood clotting time: 30mins
YELLOW  Green: 8x
 Light green: has plasma sep. gel 8x
 3-4X LIGHT BLUE
 Red glass: 0x blood clotting time: 60mins
 5X GOLD, RED PLASTIC,  Red Plastic: 5x
 Orange: 8x STAT purposes of Chemistry Blood
 NONE RED GLASS clotting time: 5mins
 Gray: 8x
 Tan: 8x
 Royal blue: 8x
 Yellow: 8x

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
ORDER OF DRAW
The order of draw as recommended by the CLSI for
both the evacuated tube system and when filling
tubes from a syringe is:

 Blood cultures (yellow stopper tubes, culture

bottles)
 Light blue stopper tubes (sodium citrate)
 Red/gray, gold stopper tubes (serum separator
tubes), red stopper plastic tubes (clot activator),
and red stopper glass tubes
 Green stopper tubes and light green (plasma
separator tubes) (heparin)
 Lavender stopper tubes (EDTA)
 Gray stopper tubes (potassium oxalate/sodium
fluoride)
 Yellow/gray or orange stopper tubes (thrombin
clot activator)

Tubes must be collected in a specific order to

prevent invalid test results caused by bacterial
contamination, tissue fluid contamination, or
carryover of additives or anticoagulants
between tubes.
 Blood drawn in a syringe is immediately
transferred to appropriate evacuated tubes to
SYRINGE prevent the formation of clots.
 Routinely used for venipuncture range from 2  It is not acceptable to puncture the rubber
to 20 mL stopper with the syringe needle and allow the
 For single draw blood to be drawn into the tube.
 For drawing blood from patients with small or
fragile veins.
The
advantage
of this
system is
that the
phlebotomi
st is able to
control the
suction
pressure on
the vein by
slowly
withdrawin
g the
syringe
plunger.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
WINGED BLOOD COLLECTION
SETS
 for performing venipuncture from very small
or very fragile veins often seen in children
and in the geriatric population
 Always hold the apparatus by the needle wings
and not by the tubing.
 or “butterflies” as they are routinely called.
 Winged blood collection needles used for
phlebotomy are usually 21 or 23 gauge with
lengths of 1/2 to 3/4 inch
 Plastic attachments to the needle that
resemble butterfly wings are used for
holding the needle during insertion and to
secure the apparatus during IV therapy.
 Safety Tip 8-8. Extreme care must be taken
when working with winged blood collection
needles to avoid accidental needle punctures.
Always hold the apparatus by the needle
wings and not by the tubing.
 If no palpable veins are found in the
antecubital area, the wrist and hand should be
examined (Fig. 10-1A).
 Tourniquets
 Vein locating devices
 70% isopropyl alcohol, iodine swabs
 2x2 inch gauze pads
 Bandage or adhesive tape
 Phlebotomy collection tray
 Slides
 Antimicrobial hand gel
 Marking pen
VENIPUNCTURE
PROCEDURE
CHECKING REQUISITION FORMS
 provide the phlebotomist with the
information needed to correctly identify the
patient, organize the necessary equipment,
collect the appropriate samples, and provide
legal protection.
 Patient’s name, age and gender
 Patient’s date of birth
 Patient’s location
 Ordering health-care provider’s name
 Tests requested
 Requested date and time of sample collection
GREETING THE PATIENT
 Phlebotomists should introduce themselves
and explain that they will be collecting a blood
sample.
PATIENT IDENTIFICATION
 The most important procedure in phlebotomy
PATIENT PREPARATION
 Positioning the Patient: supine (lying) or
sitting upright positions
 Position of the Phlebotomist: remains in the
standing position for better and greater
freedom of movement and control of the
situation
 If a fasting specimen is required, confirm that
the fasting order has been followed
17
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
TOURNIQUET APPLICATION
 maximum amount of time the tourniquet
should remain in place is 1 minute to avoid
hemoconcentration.
H-SHAPED PATTERN

 Patient Identification: CLSI, TJC, CAP require 2  includes the cephalic, median cubital, and
identifiers to ensure that blood is drawn from basilic veins in a pattern that looks like a
the right patient. slanted H.

 TOURNIQUET: This requires that the M-SHAPED PATTERN

tourniquet be applied twice during the  Includes the cephalic, median cephalic,
venipuncture procedure: 1st: vein selection median basilic, and basilic veins.
2nd: before the puncture

 -placed on the arm 3-4 inches above the

venipuncture site

SITE SELECTION
 Antecubital fossa- The preferred site for
venipuncture and is located anterior and
below the bend of the elbow.
3 MAJOR VEINS
 MEDIAN CUBITAL VEIN- vein of choice
because it is large and does not tend to move
when the needle is inserted
 CEPHALIC VEIN- usually more difficult to
locate, except possibly in larger patients, and
has more tendencies to move.
 BASILIC VEIN- the least firmly anchored; has
a tendency to “roll” and hematoma formation
is more likely to occur.
 Quite often the veins cannot be seen  The H-shaped pattern is seen in approximately
but usually felt by touching or 70 % of the population
palpating with the index finger of the  Other techniques used by phlebotomists to
non-dominant hand enhance the prominence of veins include
 They will reveal themselves as elastic massaging the arm upward from the wrist to
tubes beneath the surface of the skin. the elbow, briefly hanging the arm down, and
applying heat to the site for 3 to 5 minutes.
 If no palpable veins are found in the
 Palpation- act of locating the vein by antecubital area, the posterior side of the
sight and by touch. The ability to feel a hand, legs and feet (with physician’s approval
vein is much more important than the more susceptible to infection and the
ability to see a vein. formation of thrombi (clots), particularly in
 Veins reveal themselves as elastic tubes patients with diabetes, cardiac problems, and
coagulation disorders.)

18
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 Damaged Vein
 Hematoma
 Edema
 Burns, Scars and Tattoos
 Mastectomy
 Obesity
 IV Therapy
 Heparin and Saline Locks
 Cannulas and Fistulas
Mastectomy can be harmful to the patient and
produce erroneous test results. Removal of lymph
nodes in the mastectomy procedure interferes
with the flow of lymph fluid (lymphostasis) and
increases the blood level of lymphocytes and
waste products normally contained in the lymph
fluid.
CLEANSING THE SITE
 Cleansing is performed with a circular
motion, starting at the inside of the
venipuncture site and working outward
in widening concentric circles about 2 to
3 inches.
ASSEMBLING EQUIPMENT
EXAMINE THE NEEDLE
 visually examined for any defects such as a
non-pointed or rough (barbed) end.
ANCHORING THE VEIN
 Place the thumb 1 or 2 inches below and
slightly to the left of the insertion site and
the four fingers on the back of the arm and
pull the skin taut.
INSERTING THE NEEDLE
 bevel up, at an angle of 15 to 30 degrees
depending on the depth of the vein.
FILLING THE TUBES
REMOVAL OF THE NEEDLE
 Place folded gauze over the venipuncture
site and withdraw the needle in a smooth
swift motion. Apply pressure to the site as
soon as the needle is withdrawn.
 Never draw out the needle without
removing first the tourniquet to avoid
hematoma
DISPOSAL OF THE NEEDLE
LABELING THE TUBES
 Patient’s name and identification number
 Age and Gender of the Patient
 Date and time of collection
 Phlebotomist’s initials
CHECKING THE PATIENT’S ARM
 examine the patient’s arm to be sure the
bleeding has stopped.
 adhesive bandages/micropore tape over a
folded gauze square.
COMPLETING THE VENIPUNCTURE
PROCEDURE
 deliver the sample to the laboratory in
satisfactory condition and all appropriate
paperwork should be completed.
IMMEDIATE LOCAL COMPLICATIONS
 Localized hemoconcentration or Venous
stasis
Remedy: One minute application of tourniquet
 Syncope or Fainting
Remedy: Let the patient lie down
 Failure to obtain blood
Needle Position
Bevel Against the Wall of the Vein
Needle Too Deep/ Too Shallow
Collapsed Vein
Needle Beside the Vein
Faulty Evacuated Tube
19
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
DELAYED LOCAL COMPLICATIONS
 Thrombosis of veins- Formation of blood
clots inside the lumen of the vein due to
trauma
 Thrombophlebitis- Inflammation of the
vein due to thrombus as manifested by an
inflammatory reaction on the outer skin
surface
 Hematomas- Blue or black skin
discoloration commonly due to repeated
trauma or puncture of the veins
 General Delayed Complications- Serum
Hepatitis, AIDS
Prevention:
 Use of disposable syringe or vacutainer set
COLLECTION PRIORITIES
 Follow the procedures from the Universal  ROUTINE SAMPLES- are usually collected early
Precautions in handling infectious in the morning but can be collected
specimens throughout the day during scheduled
“sweeps” (collection times) on the floors
or from outpatients.
OTHER COMPLICATIONS
 ASAP SAMPLES -means “as soon as
Collection Attempts possible.” The response time for the
When blood is not obtained from the initial collection of this test sample is determined
venipuncture, the phlebotomist should select by each hospital or clinic and may vary by
another site. Repeat the procedure using a new laboratory tests.
needle
 STAT SAMPLES - sample is to be collected,
Nerve Injury analyzed, and results reported
Temporary or permanent nerve damage can be immediately.
caused by incorrect vein selection or improper Stat tests have the highest priority and are
venipuncture technique and may result in loss of usually ordered from the emergency
movement to the arm or hand and the possibility of department
a lawsuit.
 FASTING SAMPLE- NPO (NOTHING PER OREM);
Iatrogenic Anemia “nothing by mouth”
pertains to a condition of blood loss caused by
 FBS
treatment. An anemia can occur when large
 Lipid Profile
amounts of blood are removed for testing at one
time or over a period of time.  TIMED SAMPLES

Hemolyzed Samples  Glucose Tolerance Tests

Rupture of the red blood cell membrane releases  2-Hour Oral Glucose Tolerance Test
cellular contents into the serum or plasma and  Lactose Tolerance Test
produces interference with many test results
 BLOOD CULTURE

20
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

ARTERIAL
PUNCTURE
 Generally used for the determination of
blood oxygen, carbon dioxide tension
and blood pH (Blood Gas Analysis).
 Blood collected is called arterial blood or
oxygenated blood
 Special training is required for this
procedure
 Tourniquet is not required
 After removing the needle, apply moderate
pressure with 2 x 2 sterile gauze until
bleeding ceases
 Insert needle (still attached to syringe) in
stopper to prevent air from entering needle

A.K.A : ANAEROBIC puncture

 Radial artery
 Femoral artery (fem tap)
 Brachial artery
 Scalp artery
 Umbilical artery

 Radial artery: WRIST  most preferred

site
 Femoral artery sa singit
 Brachial artery  sa may malapit sa
median cubital vein area

 Before performing a radial artery puncture,

the Modified Allen Test is performed to
determine if the ulnar artery is capable of
providing collateral circulation to the
hand.

21
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
DERMAL PUNCTURE MAY BE
REQUIRED IN MANY ADULT
PATIENTS, INCLUDING:
 Also known as Capillary or Skin puncture  Burned or scarred patients
 Patients receiving chemotherapy who
 Blood collected by dermal puncture comes require frequent tests and whose veins
from the capillaries, arterioles, and must be reserved for therapy
venule  Patients with thrombotic tendencies
 The method of choice for collecting blood  Geriatric or other patients with very fragile
from infants and children younger than 2 veins
years.  Patients with inaccessible veins
 Obese patients
ADDITIONAL NOTES  Patients requiring home glucose
 Certain tests requires capillary blood, monitoring and point-of-care tests
such as newborn screening tests and
LANCETS
capillary blood gases.
 Drawing excessive amounts of blood  Sterile, disposable, sharp-pointed or
from premature and small infants can bladed instrument.
rapidly cause anemia, because a 2-  Punctures or cuts skin to obtain capillary
pound infant may have a total blood blood specimen.
volume of only 150 mL  Designed for either finger or heel
 A venule is a small blood vessel in the puncture.
microcirculation that allows deoxygenated
Unistik 2 Tenderfoot Lancet
blood to return from capillary beds to
larger blood vessels called veins.
FF REASONS:
 Locating superficial veins that are large
enough to accept even a small-gauge needle
is difficult in these patients.
 Use of deep veins, such as the femoral vein,
can be dangerous and may cause
complications
 Drawing excessive amounts of blood
from premature and small infants can
rapidly cause anemia, because a 2-
pound infant may have a total blood BD Microtainer Contact- Tenderlett Lancets
volume of only 150 mL Activated Lancet
EXTRACORPOREAL VOLUME
 The amount of blood outside the
patient’s body at any given time
 Should not exceed 15% of patient's total
estimated blood volume
Feather Lancet Quick Heel Lancet
 For example ang blood volume ng patient is
4,525mL
 678.75mL or 679mL  hindi dapat
lalagpas ang mawala sa katawan ng patient.

22
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 Primary danger in dermal puncture:

Accidental contact with the bone, followed
by infection or inflammation (osteomyelitis or
osteochondritis)
 To prevent contact with bone, the depth of the
puncture is critical.
 The Clinical and Laboratory Standards Institute
(CLSI) recommends that the incision depth
should not exceed 2.0 mm in a device used to
perform heel sticks.
WARMING EQUIPMENT
OSTEOMYELITIS  is an infection of the bone. Warming the site increases blood flow as much as 7
OSTEOCHONDRITIS INFECTION OF THE times
BONE BENEATH OF THE CARTILAGE
COMMERCIAL HEEL WARMER
A packet containing sodium thiosulfate and
CAPILLARY TUBES glycerin that produces heat when the chemicals are
mixed together by gentle squeezing of the packet.
 Also known as microhematocrit tubes
 small tubes used to collect approximately 50 WARM WASHCLOTHS OR TOWELS
to 75 µL of blood for the primary purpose of
performing a microhematocrit test.
 One end of tube is sealed with plastic or clay
sealants.

DERMAL PUNCTURE PROCEDURE

 Phlebotomist Preparation
 Patient Identification and Preparation
BLUE band CAPILLARY TUBE no anti-
 Patient Position
coagulant
 Site Selection
Red band heparinized
 Warming the Site
MICROTAINER TUBES  Cleansing the Site
 Performing the Puncture
 Small plastic tubes designed to hold  Puncture Device Disposal
approximately 600 µL of blood.  Sample Collection
 Color coded in the same way as evacuated  Order of Collection
tubes.  Bandaging the Patient
 Some have stoppers & markings for  Labeling the Sample
min/max fill levels.
 Completion of the Procedure
23
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 Phlebotomists should carefully examine the

information on the requisition form to
ensure that they have the appropriate
equipment to collect all required samples as
well as the skin puncture device that
corresponds to the age of the patient

 Patients for dermal puncture must be

identified using Requisition form, Verbal
identification, and ID band  Primarily required for:
 Patients with very cold or cyanotic fingers
 For heelsticks to collect multiple samples
 FOR FINGERSTICK: Patient must be seated or  For the collection of capillary blood gases
lying down with the hand supported on a
firm surface, palm up, and fingers pointed  Moistening a towel with warm water (42°C) or
downward activating a commercial heel warmer and
 FOR HEELSTICK: infants should be lying on the covering the site for 3 to 5 minutes
back with the heel in a downward position
For pediatric patients: Phlebotomist must present
 Using 70% isopropyl alcohol in a circular
a friendly, confident appearance while explaining
motion.
the procedure to the child and the parents. Do not
say the procedure will not hurt

 While the puncture is performed, the heel or

The primary dermal puncture sites:
 Plantar surface of the heel -infants
younger than 1 year old
 3rd and 4th fingers on the palmar side of the
nondominant hand-performed on adults
and children over 1 year of age.
Areas not to be punctured:
 Callused, scarred, bruised, edematous, cold
or cyanotic, or infected areas.
1. Plantar: bottom
2. Heel for baby: heel contains more tissue
than the fingers and has not yet become
callused from walking.
3. ** earlobes is not recommended.
4. ***Do not use fingers on the side of a
mastectomy.
5. Cyanotic bluish discoloration of the skin
due to lack supply of oxygen
finger should be well supported and held
firmly, without squeezing the puncture area.
Massaging the area before the puncture
may increase blood flow to the area.
Use of povidone-iodine is not recommended for
cleaning sites of dermal punctures because sample
contamination may elevate some test results,
including bilirubin, phosphorus, uric acid, and
potassium.
 Puncture device should be placed in an
appropriate sharps container.
 First drop of blood must be wiped away
with a clean gauze
24
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 Alternately applying pressure to the area
and releasing it will produce the most
satisfactory blood flow.
 Capillary Blood Gases
 Blood Smear
 EDTA Tubes
 Other anticoagulated tubes
 Serum Tubes
 Pressure is applied to the puncture site with
sterile gauze.
 Microsamples must be labeled with the
same information required for venipuncture
samples.
 Proper waste disposal of all used materials
 Proper Handwashing
 Thanking the patient and/or the parents for
their cooperation
Collection of Newborn Bilirubin
 Bilirubin is a very light-sensitive chemical
and is rapidly destroyed when exposed to
light.
INCLUDES IS DISORDERS:
 Samples must be collected quickly and
protected from excess light
 Bilirubin levels may decrease as much as
50% in a blood sample that has been
exposed to light for 2 hours.
 Require testing for as many as 29 metabolic
disorders.
 because many of these disorders cause the
buildup of unmetabolized toxic food
ingredients, it is important that the defects
be detected early in life.
The testing of newborn babies for
genetic, metabolic, hormonal, and
functional disorders that can cause
physical disabilities, mental retardation,
or even death, if not detected and treated
early.
 Levels of these substances are elevated
more rapidly in blood than urine.
 Testing for many substances is now
performed using tandem mass
spectrophotometry (MS/MS). MS/MS is
capable of screening the infant blood
sample for specific substances associated
with particular IEMs.
 Performed from blood collected by heelstick
and placed on specially designed filter
paper.
NEWBORN SCREENING
PROGRAM IN THE
PHILIPPINES:
1. CH  congenital hypothyroidism
2. CAH congenital adrenal hyperplasia
3. PKU  phenylketonuria
4. G-6-PD deficiency Glucose-6-phosphate
dehydrogenase deficiency
5. GAL  galactosemia
6. MSUD maple syrup urine disorder
25
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
CAPILLARY BLOOD GASES
 Arterial blood is the preferred sample for
blood gases (oxygen and carbon dioxide
content) and pH levels in adults
 Samples are collected in heparinized blood
gas pipettes
 The pipette should fill in less than 30
seconds.
PROCEDURE:
 Step 1. Obtain and examine the requisition
form.
 Step 2. Greet the patient and explain the
procedure to be performed.
 Step 3. Identify the patient verbally by
having him or her state both the first name
and last name and compare the information
on the patient’s ID band with the requisition
form. A parent or guardian may do this for a
child.
 Step 4. Prepare the patient and/or parents
and verify diet restrictions, as appropriate,
allergies to latex, or previous problems with
blood collection.
 Step 5. Position the patient’s arm on a firm
surface with the hand palm up. The child
may have to be held in either the vertical or
horizontal restraint.
 Step 6. Select equipment according to the
age of patient, the type of test ordered, and
the amount of blood to be collected.
 Step 7. Wash hands and put on gloves.
 Step 8. Select the puncture site in the fleshy
areas located off the center of the third or
fourth fingers on the palmar side of the
nondominant hand. Do not use the side or tip
of the finger.
 Step 9. Warm the puncture site if necessary.
 Step 10. Cleanse and dry the puncture site
with 70% isopropyl alcohol in concentric
circles and allow to air dry.
 Step ep 11. Prepare the lancet by removing
the lancet locking device and open the cap to
the microcollection container.
 Step 12. Hold the finger between the
nondominant thumb and index finger, with
the palmar surface facing up and the finger
pointing downward.
 Step 13. Place the lancet firmly on the
fleshy area of the finger perpendicular to the
fingerprint and depress the lancet trigger.
 Step 14. Discard lancet in the approved
sharps container.
 Step 15. Gently squeeze the finger and wipe
away the first drop of blood that may contain
alcohol residue and tissue fluid.
 Step 16. Collect rounded drops into micro-
collection containers in the correct order of
draw without scraping the skin. Do not milk
the site. Collect the sample within 2 minutes
to prevent clotting.
 Step 17. Cap the micro-collection container
when the correct amount of blood has been
collected.
 Step 18. Mix tubes 5 to 10 times by gentle
inversion as recommended by the
manufacturer. They may have to be gently
tapped throughout the procedure to mix the
blood with the anticoagulant.
 Step 19. Place gauze on the site and ask the
patient or parent to apply pressure until
bleeding stops
 Step 20. Label the tubes before leaving the
patient and verify identification with the
patient ID band or verbally with an
outpatient. Observe any special handling
procedures.
 Step 21. Examine the site for stoppage of
bleeding and apply bandage if the patient is
older than 2 years.
 Step 22. Dispose of used supplies in
biohazard containers.
 Step 23. Thank the patient.
 Step 24. Remove gloves and wash hands.
 Step 25. Complete paperwork.
 Step 26. Deliver sample to the laboratory
END OF PRELIM
26
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
WHAT IS URINE?
 URINE- refers to an ultrafiltrate
containing waste products excreted by
the kidneys, temporarily stored in the
urinary bladder and excreted through the
urethra.
 Normally found in the urine: sodium and
potassium ions, urea, uric acid, creatinine,
ammonia, bicarbonate ions
 Not normally found in the urine: glucose,
blood proteins, red blood cells, hemoglobin,
white blood cells (pus), bile.
CHARACTERISTICS OF URINE SPECIMEN
 Urine is a readily available and easily
collected specimen.
 Urine contains information, which can be
obtained by inexpensive laboratory tests.
URINE COLOR
 Normal daily urine output is usually 1200-
1500 mL, but range 600-2000 mL is
considered normal.
 Factors that influence urine volume:
 Fluid Intake
 Fluid loss from non renal sources
 Variations in the secretion of
Antidiuretic hormone
 Food
 Temperature
 Drugs
 Age
 POLYURIA- increase in daily urine output
 OLIGURIA- decrease in daily urine output
 ANURIA- refers to complete cessation of
urine flow
 DYSURIA- refers to painful urination
 NOCTURIA- refers increased excretion of
urine at night
 Polyuria: Greater than 2.5L/day in Adults
 Greater than 2.5 - 3mL/kg/day in
Children
 Oliguria: < 1ml/kg/hr  Infants
 < 0.5 ml/kg/hr  Children
 < 400 ml/day  Adults
 The testing of urine with procedures
commonly performed in an expeditious,
reliable, accurate, safe, and cost-effective
manner.
A ROUTINE URINALYSIS CONSISTS OF:
 Physical Examination
 Chemical Examination
 Microscopic Examination
 The microscopic portion of the urinalysis is
not a part of POCT and should not be
performed by phlebotomists.
PHYSICAL EXAMINATION OF URINE
 Urine color can range from Colorless to Black.
 metabolic functions
 physical activity
 ingested materials
 pathologic conditions
 Common color descriptions of normal urine
include pale yellow, light yellow, yellow, dark
yellow, and amber and may vary among
institutions.
27
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
PIGMENTS THAT GIVE COLOR TO
URINE
 UROCHROME- the major pigment in urine
that gives yellow color to urine which was
named by Thudichum.
 UROERYTHRIN- a pink-colored pigment,
most evident in specimen that have been
refrigerated, resulting in the precipitation of
amorphous urates.
 UROBILIN- an oxidation product of the
normal urinary constituent urobilinogen, NORMAL URINE CLARITY
imparts an Orange-Brown color to the urine  CLEAR (Midstream Clean-Catch)
that is not fresh.
PRESENCE OF TURBIDITY PROVIDES A KEY TO
THE MICROSCOPIC EXAMINATION RESULTS
 The amount of turbidity should correspond with
the amount of material observed under the
microscope.
Correlated in microscopic examination

URINE CLARITY

 Transparency/Turbidity of a urine specimen

 View in clear container (TEST TUBE) & against
newsprint

28
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
CHEMICAL EXAMINATION OF URINE
 performed using reagent strips containing
reagent impregnated test pads that test for:
-glucose -bilirubin
-ketones -specific gravity
-blood -pH
-protein -urobilinogen
-nitrite -leukocytes
 A color-producing chemical reaction occurs
when the reagent pads come in contact with
urine.
 The color reaction can be read visually by URINE SPECIMEN AND COLLECTION
comparing the strip against a color chart on the  CONTAINERS:
container.
 Clean, dry, leak-proof containers
 Wide-mouth and wide flat bottom
 Clear
 Recommended capacity: 50mL
 Disposable containers should be used to
eliminate the chance of contamination

URINE CONTAINERS

 Glu and Bil – 30 seconds
 SpGr – 45 seconds
 Ketones- 40 seconds
 PPBUN – 60 seconds
 Leukocytes- 120 seconds
 That is the specified amount of time for the
reaction to occur
 *Failure to read the reactions within the
time frame specified by the manufacturer
will also produce inaccurate results.
 4 basic parameter GLUCOSE, PROTEIN,
Ph, S.G
 11TH PARAMETER= ASCORBIC ACID- 40
SECONDS
URINE COLLECTION
 LABELS:
 Patient’s name and identification
number
 Patient’s age and sex
 Date and time of collection
 Labels must be attached to the container, not
to the lid, and should not become detached if the
container is refrigerated or frozen.
REQUISITION:
 A requisition form must accompany specimens
 Information on the form must match the
information on the specimen label
 Method of collection or type of
specimen
 Possible interfering medications
 Patient’s clinical information
 Time the specimen is received in the
laboratory
29
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
through the urethra into the bladder. This
type of sample is collected on a patient
unable to void, babies, or bedridden patients.
 Improperly labeled and collected specimens
 MIDSTREAM CLEAN-CATCH: Patients are
should be rejected by the laboratory
provided with sterile containers and antiseptic
 Specimens in unlabeled containers
materials for cleansing the genitalia.
 Nonmatching labels and requisition forms
 SUPRAPUBIC SAMPLE: external introduction of
 Specimens contaminated with feces or toilet
a needle through the abdomen into the
paper
bladder.
 Containers with contaminated exteriors
 Specimens of insufficient quantity
 Specimens that have been improperly
transported

URINE SPECIMEN HANDLING

SPECIMEN INTEGRITY

 Following collection, specimens should be

delivered to the laboratory promptly and tested
within 2 hours.

 A specimen that cannot be delivered and tested

within 2 hours should be refrigerated or have
an appropriate chemical preservative added.

TYPES OF URINE SPECIMEN

URINE DRUG SAMPLE COLLECTION

 RANDOM: most commonly received specimen;
collected anytime of the day
 FIRST MORNING/ 8 HR SPECIMEN: Concentrated
specimen
 24-HR: Patients are provided with large plastic
containers that may contain a preservative. To
obtain an accurate timed sample, it is
necessary for the patient to begin and end the
collection period with an empty bladder.
 CATHETERIZED: collected under sterile
conditions by passing a sterile hollow tube
 For urine samples to withstand legal scrutiny, it
is necessary to prove that no tampering (e.g.,
adulteration, substitution, or dilution) took
place.
 If a witnessed sample collection is ordered, a
same-gender collector will observe the
collection of 30 to 45 mL of urine. Witnessed
and unwitnessed collections should be
immediately handed to the collector.

30
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

FECES / STOOL
 SOFT AND WATERY – Diarrhea
 Solid/ semi-solid body waste discharged
from the large intestine through the anus  HARD AND SCYBALOUS (LIKE GOAT
during defecation. DROPPINGS)- Spastic constipation
 BULKY FROTHY STOOL- Bile duct obstruction,
COMPONENTS OF A NORMAL STOOL
Pancreatic disorders, abundant fats
 Bacteria  RIBBON-LIKE STOOL – Intestinal constriction
 Undigested food stuff due to malignancy (colon cancer)
 Bile pigments (Urobilin/ Stercobilin)  MUCUS AND BLOOD-STREAKED STOOL –
 Water and electrolytes Amoebic colitis, Dysentery, Malignancy
 Enzymes
 Trypsin
 RICE WATERY STOOL- cholera
 Chymotrypsin
 Aminopeptidase
 Lipase
 Amylase

 Volume: 100-200 grams/day

 Color: Light to Dark Brown
 Odor: Offensive (due to Indole and Skatole)
 Form and Consistency: Soft and well formed
REACTION:
 Alkaline: Increase protein diet
 Acidic: Increase vegetable, CHO, and fats in the
diet
ABNORMAL VARIATION IN
COLOR
FECAL SAMPLE COLLECTION
 BLACK- upper gastrointestinal bleeding, intake
iron, etc.  Detailed instructions and appropriate
 RED- bleeding in lower GIT, intake of beets, food containers should be provided.
coloring, Rifampin and Pyridium compounds.  Containers for collection of fecal (stool)
 PALE YELLOW, WHITE, GRAY- Bile duct obstruction, samples:
Barium intake  Random samples used for cultures, ova and
 GREEN- Presence of biliverdin, Oral antibiotics, parasites (O&P), microscopic examination
Green Vegetables for cells, fats and fibers, and detection of
blood are collected in cardboard

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
containers with wax-coated interiors or iodine by rotary motion using the applicator
plastic containers with wide openings and stick.
screwcapped tops similar to those used for
 Formed stool: portion of stool from area to
urine samples.
include inside and outside of the specimen
 Stool with mucus: small portion of mucus and
mix
 Watery stool: if mucus is not present, small
portion of stool
Cover the saline/ iodine with cover slip

CHEMICAL EXAMINATION OF STOOL

Bedpan: For quantitative testing, such as for fecal
fats collected within 3-day collection.
 The stool collected should only be about thumb
size/ pea size of forming or soft specimen and
5-6 spoonful of watery specimen.
 For specimen with no preservatives, tighten
the lid to prevent leakage and refrigerate the
sample if will not be transported
immediately.
 Patients should understand that the specimen
must not be contaminated with urine, toilet
water, fibers, gauze threads, and tissue
papers.
SPECIMEN: FECAL OCCULT BLOOD
TESTING (FOBT)
 Hidden Blood, not seen by microscopic
examination.
 Normally found in small amount, 2.5mL/ 150
grams of stool
 Screening test for colorectal cancer and GIT
bleeding.
Based on the pseudoperoxidase
activity of hemoglobin molecule reacting with
chromogen.
>2.5mL/ 150 grams of stool = SIGNIFICANT

PRESERVATION OF STOOL
 Gum Guaiac
 Refrigeration  Benzidine
 Chemical Preservative  Ortho toluidine
 Sodium Acetate
 Formalin Positive Result: BLUE CHROMOGEN
 Merthiolate Iodine Formaldehyde
 NSS w/ glycerine
DIETARY RESTRICTIONS 3 DAYS
 Polyvinyl Acohol BEFORE THE EXAMINATION (FOBT)
 Red Meat
 Horse radish
 Melons
 Raw broccoli
 Cauliflower
 1 drop of Normal Saline Solution/ 1% Lugol’s  Turnip
Iodine Solution on a glass slide. Place a small  Vitamin C
portion of stool specimen to the drop of solution.  Aspirin and NSAIDS- restriction of 7 days
Make a smooth emulsion in the drop of saline/ prior specimen collection

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 Many phlebotomists are employed by FOUR TRADITIONAL HOSPITAL SERVICES

hospitals.

 Other employment settings include physician

office laboratories (POLs), health
maintenance organizations (HMOs),
reference laboratories, urgent care centers, Professional Nursing
nursing homes, home health-care agencies, services services
clinics and blood donor centers.

HOSPITAL ORGANIZATION
 Hospitals are classified in different terms such
as community hospitals, teaching hospitals
(university-based), and nonprofit and for-
profit hospitals. There are hospitals that
specialize in a particular type of patient or
illness, such as children, mental health,
rehabilitation, and cancer treatment.
 The traditional hospital contains many
different patient areas and departments to
which the phlebotomist must travel to collect
samples.
Fiscal Support
services services
NURSING SERVICES
 This service deals directly with patient care.
 Members associated with this service are
registered nurses (RNs), licensed practical
nurses (LPNs), certified nursing assistants
(CNAs), and the unit secretary.
 This service consists of the cardiac care unit
(CCU), central supply, emergency
department (ED), hospital patient-care units,
infection control, intensive care unit (ICU),
nursery, social services, and the operating
room (OR) where phlebotomists interact
most often with this service.

SUPPORT SERVICES
 Maintain the hospital and include
communications systems, food
service/dietary,
housekeeping/environmental services,
laundry, engineering and maintenance, and
security.
 Mainly for the maintenance of the hospital.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

FISCAL SERVICES
 Fiscal services manage the business aspect of
a hospital.
 These services include:
- Accounting
- Admitting
- The business office
- Credit and collection  Radiology department uses various forms of
- Data Processing radiant energy to diagnose and treat disease.
- Health information management  The allied health-care professional in this
- Planning, and public relations departments department is a radiographer or radiologic
that include marketing and outreach technologist.
programs.  A radiologist, a physician that administers
diagnostic procedures and interprets
radiographs.

 This department uses high-energy x-rays or

ionizing radiation to stop the growth of
cancer cells.
 These procedures are performed by Radiation
Therapy Technologists.

 uses the characteristics of radioactive

substances in the diagnosis and treatment of
disease. Radioactive materials, called
PROFESSIONAL SERVICES radioisotopes, emit rays as they disintegrate,
and the rays are measured on specialized
 This service consists of the departments of instruments.
the hospital that assist the physician in the  Nuclear medicine technologists perform
diagnosis and treatment of disease. these procedures under the supervision of a
 Main departments in this service include: physician.
- The Clinical Laboratory  Two types of tests are used:
- Radiology/Medical imaging  In vitro tests analyze blood and urine
- Radiation Therapy samples using radioactive materials to
- Nuclear Medicine detect levels of hormones, drugs, and
- Occupational Therapy other substances.
- Pharmacy  In vivo tests involve administering
- Physical Therapy radioactive material to the patient by
- Respiratory Therapy intravenous (IV) injection and
- Cardiovascular Testing measuring the emitted rays to
examine organs and evaluate their
function.
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 teaches techniques that enable patients with
physical, mental, or emotional disabilities to
function within their limitations in daily
living.
 Occupational therapists and technicians
provide this instruction.
 The pharmacy department dispenses the
medications prescribed by physicians.
 Persons trained to dispense medications are
called pharmacists who may be assisted by
pharmacy technicians.
 provides treatment to patients who have
been disabled as a result of illness or injury
by using procedures involving water, heat,
massage, ultrasound, and exercise.
 Physical therapists and physical therapy
assistants are the professionals trained to
provide this therapy.
 provide treatment in breathing disorders and
perform testing to evaluate lung function,
performed by Respiratory Therapists.
 RTs may also perform the arterial punctures
used to evaluate arterial blood gases.
 Cardiac technicians under the supervision of a
cardiologist evaluate cardiac function using
electrocardiograms, stress tests, and
imaging techniques.
 clinical laboratory provides data to the
healthcare team to aid in determining the
diagnosis, treatment, and prognosis of a
patient.
PHYSICIANS OFFICE
LABORATORIES (POLS) AND
GROUP PRACTICES
 A diagnostic lab in a physician's office with an
abbreviated menu of tests.
 -That can be performed while the physician is
in the office, and the patient will have a better
care from the physician.
HEALTH MANAGEMENT
ORGANIZATIONS (HMOS)
 a group or an organization of medical
insurance providers that limit coverage to
medical aid provided from doctors that are
under the contract of HMO.
 Members are charged a prepaid fee for all
services performed during a designated time
period.
REFERENCE LABORATORIES
 Large, independent reference laboratories
contract with health-care providers and
institutions to perform both routine and
highly specialized tests.
 Phlebotomists are hired to collect samples from
patients referred to the reference laboratory.
They may be stationed at the laboratory or at
off-site designated collection facilities.
Phlebotomists also may be assigned to
process samples received in the reference
laboratory from its contracted outside
health-care facilities.
GOVERNMENT AND HOSPITAL
CLINICS
 Hospital-sponsored specialty clinics, such as
cancer, urology, and pediatric clinics, provide
more cost-effective delivery of health care to
more patients.
 Increased emphasis on preventive medicine and
alternative medicine has resulted in the
establishment of wellness clinics for health
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
screening. Phlebotomists may be employed in ACID HEMATIN FOR HEMOGLOBIN
these settings.
DETERMINATION
HOME HEALTH CARE
 Home Health Care includes any professional
support services that allow a person to live
safely in their home. A Medical Care provided
in a patient's home.

 Home health care can include broad care given

by skilled medical professionals, including
skilled nursing care, physical therapy,
occupational therapy and speech therapy.
Home health care can also include skilled, non-
medical care, such as medical social services
or assistance with daily living from a highly TWO TEST FOR HEMOGLOBIN DETERMINATION
qualified home health aide. MANUAL: DRABKIN’S CYANMETHEMOGLOBIN
METHOD
 here we used Drabkin’s Reagent. It is used for
quantitative choloremetric determination of
HEMATOLOGY SECTION hemoglobin concentration in whole blood.
 Complete Blood Count  PRINCIPLES: This method lies in conversion of
 Hemoglobin hemoglobin. Icoconvert yung hemoglobin na
 Hematocrit nakita sa whole blood to cyanmethehemoglobin
 Red Blood Cell Count by the addition of postassium cyanide and ferri
 RBC indices cyanide. With the absorabance value of 540
-Mean Corpuscular Volume nanometers. So we will use photoelectric
-Mean Corpuscular Hemoglobin calorimeter or spectrophometer.
-Mean Corpuscular Hemoglobin
ACID HEMATIN
Concentration
PRINCIPLES:
 White Blood Cell Count
 The blood is mix with normal hydrochloric acid.
 WBC Differential Count
In the conversion of hemoglobin to acid hematin
 Platelet (PLT) count
which is a brown color.
 Erythrocyte sedimentation rate (ESR)
 In the container our solution is same color.
 Reticulocyte (Retic) count
 Then we will now see the values of how much
HEMOGLOBIN the hemoglobin.
 Identified by Felix Seyler and proved that this
HEMATOCRIT
was the true coloring matter of the blood
 Upon centrifugation of blood, the volume of the
 An iron-bearing protein, hemoglobin’s main
red blood cells represents the packed red cell
function is to transport oxygen from the lungs to
volume or hematocrit
tissues and transport carbon dioxide from the
 Is done to measure the RBCs in blood.
tissues to the lungs for exhalation.
 Also known as packed cell volume.
HEMOGLOBIN DETERMINATION  Measure of the ratio of the volume occupied by
 Manual: Drabkin’s Cyanmethemoglobin Method RBC.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
MICROHEMATOCRIT PLATELET COUNT
 Direct Platelet count
 Indirect Platelet count
To avoid reading it repeatedly, in zigzag motion
starting from the bottom to up. (left to right first).

CLINICAL CHEMISTRY SECTION

 Glucose Test (FBS, RBS)
 Lipid Profile
Cholesterol
LDL
HDL
Triglycerides (TAG)
 Determination of packed cell volume or  Liver Function Test
hematocrit using a small quantity of whole SGPT
blood. SGOT
RED BLOOD CELL Bilirubin
 Also known as erythrocytes, they are small cells ALP
shaped like biconcave discs  Kidney Function Test
 Primary function is to ferry oxygen in blood to BUN
all cells of the body Creatinine
WHITE BLOOD CELL  Enzyme testing
 Also known as leukocytes, the white blood cells
help defend the body against damage by
bacteria, viruses or parasites

Granulocytes
Gram Stain /
• Neutrophils Culture and
Sputum
AFB staining
coiling
• Eosinophils Sensitivity
• Basophils
Agranulocytes GRAM STAIN
• Lymphocyte  The most commonly used differential stain in
• Monocyte clinical microbiology laboratory
 It is a complex and differential staining
 Granulocytes – means have granules. procedure use to differentiate the gram positive
 Agranulocytes –doesn’t have granules. and gram negative bacteria.
STAINING OF BLOOD SMEAR  Gram positive contains thick layers of
 Wright stain peptidoglycan. Stain purple.
How to Read:  Gram Negative thin layer of peptidoglycan and
 Need to do Blood Smear. stain pink.
 Usage of Wright Stain
PLATELETS
 Platelets are needed for the clotting process that  Crystal Violet
occurs when blood vessels are ruptured or  Iodine / Gram’s Iodine
broken  Acid Alcohol/ alcohol
 Safranin
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 Smear of bacteria.
 Stain the smear with crystal Violet. This is the
primary stain. Stay for 1 minute.
 Rinse to running water or ilulubog lang.
 Iodine then stay 1 minute. Rinse again.
 Acid Alcohol. Decolorizer.
 Then wash again.
 Add the safranin.
SPUTUM SMEAR
ANTIMICROBIAL SUSCEPTIBILITY
TESTING
 Used to identify the susceptibility patterns of the
bacterial isolate to the antimicrobial agents
 Determine what particular antibiotic or anti-
fungal drugs to stop growth of bacteria or fungi.
CULTURE AND SENSITIVITY
 Pag maliit yung hallow zone is the bacteria is
resistance doon sa antibiotic.
 Pag nakakapatay, susceptible ang bacteria.
 It allows rapid and reliable identification of
patient with tuberculosis or pneumonia.
 Gumagawa ng Sputum Coiling.
 Choose a new and clean slides
 Label the slides.
 Use wood applicator stick.
 Collect the thick or greenish part.
 Evenly distribute the specimen.
 Allow to dry.
 Then Heat Fix – para di mawashout ang
specimen.
- Bunsen Burner
- Alcohol Burner
- Electrical Heating Block.
 If using burner pass the slide 2 to 3 times.
 If using Electrical Heating Block (-2 Hours, 65C –
70C)
 Main aim in staining: Differentiate bacteria into
acid fast group and non-acid fast group by the
use of stain.
 By the use of this stain: Ziehl- Neelsen Staining
- Carbol-Fuschin (Primary Dye)
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
- 20% Sulphuric Acid or Acid Alcohol IMMUNOLOGY AND SEROLOGY SECTION
(Decolorizer)
- Methylene Blue Dye (Counterstain) or  HbsAg testing
Malachite Green  C-reactive Protein
 Carbol Fuschin for about 3 mins. (5-10 mins)  Anti-HIV Testing
 Rinse and Shake
 Destain with Acid Alcohol. Drop and drop until
HISTOPATHOLOGY SECTION
the color remove.  Pap Smear
 Rinse and Shake Again.  Biopsy
 Methylene Blue Dye for 2 minute.  Cytological Testing
 Rinse Again and Shake.
 Blot Gently with Kimwipe.
 Acid Fast bacilli will be red.
 Heating.

 Laboratory Director (Pathologist)

 Laboratory Manager (Administrator)
 Technical Supervisor
 Medical Laboratory Scientist
 Medical Laboratory Technician
 Laboratory Assistant
 Phlebotomist

ADDITIONAL NOTES

 LUNG CENTER OF THE PHILIPPINES (Quezon City)

MICROBIOLOGY SECTION – They cater clinical chemistry samples.

 Culture and Sensitivity  EAST AVENUE MEDICAL CENTER- For Drug

 Gram Stain Testing and Water Bacteriology.
 AFB
 KOH preparation  NATIONAL KIDNEY AND TRANSPLANT INSTITUTE –
For Hematology samples.
CLINICAL MICROSCOPY SECTION
 Urinalysis (UA)
 RESEARCH INSTITUTE FOR TROPICAL MEDICINE –
 Fecalysis (FA)
for microbiology and parasitology samples.
 Occult Blood Testing

BLOOD BANKING SECTION  PHILIPPINE HEART CENTER – For cardiovascular

 Blood Typing
 Crossmatching
 Antibody Screening and Identification
 Direct Coomb’s Test and Indirect Coomb’s Test
disease.
 SAN LAZARO HOSPITAL – For HIV, AIDS, Sexually
Transmitted Disease, Hepatitis.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

CODE OF ETHICS
 Principles of right and wrong
 Provide the personal and professional rules of
performance and moral behavior as set by
members of a profession.
BIOETHICS/ MEDICAL ETHICS
 focus on the patient to ensure that all members
of a health-care team possess and exhibit the
skill, knowledge, training, professionalism,
and moral standards necessary to serve the
patient.
THE PATIENT’S BILL OF RIGHTS
 Specified what the patient has a right to expect
 Failure to respect patients’ rights and
during medical treatment.
performing beneath the standard of care can
result in legal action initiated by the patient or
the patient’s family.
 Penalties may include revocation of
professional licenses, monetary fines, or
imprisonment.

 The Patient’s Bill of Rights- A document first
published by the American Hospital
Association (AHA) in 1973
 A patient’s rights and dignity must be
protected in the process of providing quality
care.
THE PATIENT CARE
PARTNERSHIP
 In 2003 the AHA published a revision to the
Patient’s Bill of Rights titled The Patient Care
Partnership: Understanding Expectations,
Rights and Responsibilities.
 A major goal of the revision was to provide
information in plain, straightforward language
to better communicate with patients.
 AHA-American Hospital Association
PLAINTIFF
 The person who brings the lawsuit or action.
DEFENDANT
 the health-care worker or institution against
whom the action or lawsuit is filed.
CRIMINAL LAWSUIT
 is an action initiated by the state for
committing an illegal act against the public
welfare and can be punishable by
imprisonment.
CIVIL LAWSUIT
 is a court action between parties seeking
monetary compensation for an offense.
 Plaintiff: Someone who makes a legal
complaint against someone else in court
 Civil: consists of a monetary award and never
consists of imprisonment
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 A wrongful act committed by one person

against another that causes harm to the person
or his or her property is called a tort.
CLASSIFICATION OF TORT
1. INTENTIONAL TORT
 Assault
 Battery
 Defamation
2. UNINTENTIONAL TORT
 Negligence
 Malpractice

The concept of this area of law is to redress a wrong

done to a person and provide relief from the wrongful
acts of others, usually by awarding monetary
damages as compensation. The original intent
of tort is to provide full compensation for proved
harms.
 MALPRACTICE - is misconduct or lack of
skill by a health-care professional that results
in injury to the patient.
 NEGLIGENCE- which is defined as failure to
give reasonable care by the health-care
provider, must be proven in a malpractice
suit.

 ASSAULT is the threat to touch another person

without his or her consent and with the
intention of causing fear of harm.
 BATTERY is the actual harmful touching of a
person without his or her consent.
 DEFAMATION is spoken or written words that
can injure a person’s reputation.
- Libel- false defamatory writing that is
published.
- Slander - false and malicious spoken
word.
- Invasion of privacy - the violation of the
patient’s right to be left alone and the right
to be free from unwanted exposure to
public view.
Libel: through written words (published)
Slander: through verbal actions
 DUTY: Indicates that there was an established
standard of care and proof that it was not
followed. The standard of care can be for the
procedure and for the training and
evaluating protocols of the phlebotomist
performing the procedure.
 BREACH OF DUTY: The plaintiff (patient)
must show what actually happened and that
the defendant (phlebotomist) failed to
perform.
 CAUSATION: Indicates that the breach of
duty directly caused the injury and that no
other factors could have contributed.
 DAMAGES: Actual physical, emotional, or
financial injury had to occur to the plaintiff
(patient) because of the negligent act.

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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
LABORATORY MEDICINE
 Refers to the discipline involved in the
selection, provision, and interpretation of
diagnostic testing.
 Analytical testing
 Research
 Administration
 Teaching Activities
 Clinical service
 Study of biochemical processes associated
with health and disease.
 Measurement of constituents in body fluids or
tissues to facilitate diagnosis of disease.
 Monitor the effect of treatment and measuring
the drug levels in blood and other body fluids.
TOOLS AND METHODS
 Theory and Practice of Reference Intervals
 Use of Internal Quality Control and Proficiency
Testing
 Introduction of Automation
 Concepts of Diagnostic Testing
UNITS OF MEASURE
 Reporting of laboratory results is often
expressed in terms of substance concentration
(e.g., moles) or the mass of a substance (e.g.,
mg/dL, g/dL, g/L, mEq/L, and IU) rather than
in SI units.
 Analytes must be reported using moles of
solute/volume of solution (mmol/L).
 Liter is used as the reference volume.
PATIENT PREPARATION
FASTING
HEMOLYZED SPECIMEN
 Fasting requirement is between 8-12 hours
 Fasting specimen: FBS, GTT, Lipids,
lipoproteins, gastrin and insulin
DIET
 Metabolic products of food can increase in
venous blood.
 The tests most affected are glucose and
triglycerides
 Clinical chemistry tests are performed primarily
on serum collected in gel barrier tubes.
 Tests are also performed on plasma, CSF,
 Urine, Synovial fluid, Pleural Fluid,
Pericardial Fluid, Peritoneal Fluid.
 Serum and plasma are obtained by
centrifugation (for 10mins).
 For acquiring serum, blood samples
must be allowed to clot fully (takes
about 20–30 minutes) before
centrifugation to ensure complete
separation of the cells and serum.
 Serum or plasma should be separated
from cells as soon as possible.
 Ideally, all measurements should be
performed within 1 hour after collection.
STORAGE AND TRANSPORT OF
SPECIMENS
 Serum or plasma must be stored at 4° C to 6 °
C or;
 Freeze at -20 ° C, if analysis is to delayed by
more than 4 hrs.
 During storage (ambient temperature,
refrigeration or freezing), the concentration
of a blood constituent in the specimen may
change as a result of various processes,
including adsorption to glass or plastic tubes,
protein denaturation, water movement into
cells resulting in hemoconcentration of serum
and plasma, etc.
 Appear red because of the release of
hemoglobin from RBCs
 Severe hemolysis causes a slight dilutional
effect on the analytes present in serum or
plasma.
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
ICTERIC SPECIMEN COLORIMETRY
 Appear yellow because of the presence of KINDS OF COLORIMETRY
excess bilirubin.
LIPEMIC SPECIMEN  Visual Colorimetry – uses the eyes in
determining end point
 Appear cloudy because of increased lipids.
 Photoelectric Colorimetry

2 CATEGORIES
 Spectrophotometry (Spectrophotometric
measurements)
 Filter photometry (Photometric
measurements)

BEER’S LAW OR BEER-

LAMBERT’S LAW
IT IS FOLLOWED ONLY IF THE FOLLOWING
CONDITIONS ARE MET:

 Incident radiation on the substance of interest

1. SPECTROMETRY
- spectrophotometry, atomic absorption,
and mass spectrometry
2. LUMINESCENCE
- fluorescence, chemiluminescence, and
nephelometry
3. ELECTROANALYTIC METHOD
- electrophoresis, potentiometry, and
amperometry
4. CHROMATOGRAPHY
- gas, liquid, and thin-layer
 The spectrophotometer technique is to
measure light intensity as a function of
wavelength.
 an appearance of cold body radiation. ,
LUMINESCENCE is the low-temperature
emission of light
 Electroanalytical methods exploit
relationship between electricity and
chemistry to characterize a sample.
 a physical method of separation that
distributes components to separate between
two phases, one stationary (stationary phase),
the other MOBILE PHASE
is monochromatic.
 The solvent absorption is insignificant
compared with the solute absorbance.
 The solute concentration is w/in given
limits.
 An optical interference is not present.
 A chemical reaction does not occur between
the molecule of interest and another solute or
solvent molecule.
TRANSMITTANCE
 Ratio of the radiant energy transmitted,
divided by the radiant energy incident on the
sample.
 %T = It/Iox100
ABSORBANCE
 The amount of light absorbed
 Proportional to the inverse log of
the
transmittance
 Mathematically derived from %T
 A = 2 – log%T
 A = -log%T = 1/log%T
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MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 The piston does not come in contact with the
liquid.
POSITIVE DISPLACEMENT PIPETTE
 operates by moving the piston in the pipette
tip or barrel.
DISPENSER/DILUTOR
 obtains liquid from a common reservoir.

Spectrophotometer measures light absorption as a

function of wavelength in UV as well as visible
regions and follows the Beer Lambert’s law of light
absorption.

BASIC COMPONENTS
 Light source
PIPETTE TIPS
 Device to isolate light of a desired wavelength-  Designed for use with air-displacement
Monochromator pipette.
 Cuvette  Ensure that pipette tip is seated snugly onto the
 Photodetector end of the pipette and free from any
 Read-out device deformity.
 Data system  Positive displacement pipettes use tips made
up of straight columns of glass or plastics.
AUTOMATIC PIPETTE (MACRO: > 1 ML;
MICRO: < 1 ML)
ADVANTAGES
 time saving
 safety
 stability
 ease of use
 increase in precision
 lack of required cleaning
 tips (contaminated) are often disposable
TYPES:
 Air Displacement
 Positive Displacement
 Dispenser/ Dilutor

AIR-DISPLACEMENT PIPETTE
 relies on a piston for creating suction to draw
the sample into a disposable tip

44
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 Insert the test strip into test strip slot until it will
go no further with gold-colored bars printed
“SD- Check” facing up and toward the meter.
 The meter turns on automatically. Check that the
code number on the meter matches the code
on the container of test strips you are using.
 Obtain a drop of blood sample using the lancet
and lancing device.
 Touch and hold drop of blood to the tip of the
strip.
 The blood will be drawn into the strip
automatically. Hold your finger to the edge of the
strip until the yellow window is completely
filled with blood.
POINT-OF-CARE TESTING: BLOOD GLUCOSE  When blood is applied to strip, the display
MONITORING counts down from 1 to 5 second and your
PROCEDURE: result appears on the display in just 5
seconds.\
TESTING BLOOD GLUCOSE  Remove and discard the used test strip.
 Remove a new strip from container. Be sure
to tightly replace container cap after
removing test strip.
END OF MIDTERM 45
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
MICROBIOLOGY SECTION
 Responsible for the identification of
pathogenic microorganisms and for hospital
infection control.
DIVIDED INTO
 Bacteriology
 Mycology
 Virology
 Parasitology
As part of Laboratory Safety, CDC has classified
microorganisms into various biosafety categories.
 BSL 1- no known pathogenic potential for
immunocompetent individuals, NO RISK.
 BSL 2 – this category includes the most common
microorganisms associated with laboratory
acquired infections; MODERATE RISK
 BSL 3- Mycobacterium tuberculosis, Brucella,
Coccidiodes immitis, Rickettsiaea and
Arboviruses
 BSL 4- used in research facilities includes a
limited of exotoxic viruses Filovirus and
Arenavirus.
BSL 1: Example of microorganism: Bacillus subtilis
BSL 2: Staphylococcus, HBV, HIV, enteric
pathogens like Salmonella and Shigella
CHEMICAL GERMICIDE CATEGORIES
A.) STERILIZATION
 complete destruction or removal of all living
forms, including their spores
MOIST HEAT
 Autoclave- ***steam under pressure, 121°C
at 15 psi for 15- 30minutes
 Tyndallization- 100°C for 30mins for 3
consecutive days
 Pasteurization- 63°C for 30 mins; 72°C for 15
seconds
 Boiling- 100°C for 15-30 minutes
DRY HEAT
 a. Oven- 160-180°C for 1-2 hrs, for glasswares
 b. Incineration- most common for disposal of
infectious wastes - 870 - 980°C
 c. Flaming- for loops and needles
 d. Cremation- burning materials into ashes, to
control disease.
FILTRATION
 a. Seitz filter- used to filter heat labile fluid
 b. Membrane filter or cellulose filter
B.) DISINFECTION
 to destroy or irreversibly inactivate bacteria,
viruses and fungi but not necessarily the
spores on inanimate object.
ANTISEPTIC
 70% ethyl/ isopropyl alcohol
 Iodophors (iodine + detergent)
 Chlorhexidine
 Hexachlorophene
DISINFECTANTS
 Sodium Hypochlorite
 QUATS- Quaternary Ammonium Compounds
46
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
URINE
 Specimen of choice for bacterial culture is
Midstream Clean Catch
BLOOD  *Catheterized for those unable to produce
 Avoid the normal skin flora while collecting  *Suprapubic urine for anaerobic culture
blood for culture
 *Escherichia coli- major cause of UTI
 Most common anticoagulant for blood  *Staphylococcus saprophyticus- cause of UTI
collection: ***Sodium Polyanethol Sulfonate among young females
(SPS)
STOOL
CSF  Used for the detection of enteric pathogen
 Samples are collected in sterile tubes usually GENITAL TRACT SPECIMENS
numbered 1 through 3, representing the order  To detect presence of sexually transmitted
in which the sample was collected: pathogen
 Tube No. 1 is designated for *chemistry.
 Tube No. 2 is designated for *microbiology.
 Tube No. 3 is designated for *hematology.
 Tube No. 4 for additional testing
(Microbiology, chemistry, serology or
hematology)
THROAT AND NASOPHARYNGEAL SWABS
 Specimen of choice to detect carrier state of
Neisseria meningitidis and to detect presence
of Bordetella pertussis and Haemophilus
influenzae
SPUTUM
 May often be contaminated with normal flora
so it is important to evaluate the quality of
specimen.
 Note the number of Squamous epithelial cells/
LPF and PMNs to evaluate acceptability of
specimen.
 Collected ideally in morning when it is most
concentrated.
CHARACTERISTICS OF BACTERIA
 Prokaryotic
 Has both RNA and DNA
 Measured in µm
 Requires microscopy for visualization
 Average size 0.4-2.0µm
 Produce either exotoxin or endotoxin
Exotoxin - Associated with gram positive bacteria
Endotoxin - Associated with gram negative bacteria

47
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
COMMON STRUCTURAL COMPONENTS OF
BACTERIAL CELL
 CELL WALL  also referred to as the
*peptidoglycan layer or murein layer
 CELLULAR APPENDAGES aside from a cell wall
of bacteria, some bacteria contains additional
structures like:
 FLAGELLA
 are complex structures, mostly GENERAL FEATURES OF FUNGI
composed of the protein “flagellin”.
 Responsible for bacterial motility  Eukaryotic cell
 CAPSULE  Most are obligate aerobes
 referred to as the slime layer.  Fungi can be stained using gram staining
 Protects bacteria from attack by procedure.
cells of the human defense system.  Saprophytic and heterotrophic
 FIMBRIAE OR PILI  A chlorophyll
 are hairlike, proteinaceous  Presence of Chitin and glycan as component of
structures that extend from the cell cell wall
membrane into the external  Ergosterol is the steroid molecule that replaces
environment. cholesterol found in the cell membrane of an
Two types common pili and sex pili animal cell.
GRAM STAINING SEXUAL AND ASEXUAL REPRODUCTION OF
 The most fundamental test used in bacterial FUNGI
identification schemes.
 Separates almost all medically important  Asexual reproduction: happens when a single
bacteria into two general types: parent cell divides to form a daughter cell via
 Gram positive bacteria mitosis.
 Gram negative bacteria  Production of either of the following
 Reactions of bacteria with gram stain depends asexual spores:
on the ability of bacterial cell wall to trap  sporangiospore, blastospore,
certain dyes, that even after decolorization, the conidiospore, arthrospore and
primary stain is stuck or trap inside the cell chlamydospore
wall.  Sexual reproduction: happens when two
PURPOSE REAGENT Gram Gram similar or even dissimilar morphology/fungi
Positive Negative fuse together.
 Production of either of the following
Primary Crystal Violet Violet sexual spores:
Stain/ Initial Violet  Zygospore, basidiospore,
Stain ascospore
Mordant Gram’s Violet Violet
Iodine

Decolorizer 95% Violet colorless

Alcohol

Secondary Safranin Deep Blue- Red/pink

Stain Violet/purple

48
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 They cannot make food, take in food, or
produce wastes
 Viruses are non-cellular particles made up of
genetic material and protein that can invade
living cells.
 Contains DNA or RNA
 Contain protein coat
 Some are enclosed by an envelope
 Some viruses have spikes
 Most viruses infect only specific types of cells
in one host
 Host range is determined by specific host
 Superficial mycoses attachment sites and cellular factors.
 Cutaneous mycoses
 Subcutaneous mycoses
 Systemic mycoses
 Opportunistic

 Respiratory Tract Secretions

 Cerebrospinal Fluid
 Blood CHARACTERISTICS OF PARASITE
 Hair, Skin and Nail scrapings  Parasites are non producers (they feed off
 Urine their host)
 Tissue, Bone Marrow and Sterile Body fluids  They can be classified based on their location
in the host body, namely: ectoparasites,
endoparasites and mesoparasites.
 Endoparasites are parasites that live
 Direct Microscopic Examination inside the body of the host
 Nucleic Acid amplification  Ectoparasite are parasites that live
 Cultivation on the outer surface of the host and
 Serodiagnosis generally attach themselves during
feeding
 Mesoparasite These are the parasites
that live relatively embedded in its host.
CHARACTERISTICS OF VIRUS They penetrate external openings,
 Does not belong to any kingdom. like the buccal cavity or the external
ear.

49
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
DEHYDRATION
 Removal of water from tissue in preparation
for tissue impregnation
 Microscopic study of the tissue affected by
disease.
CLEARING (DE-ALCOHOLIZATION)
 Intermediate process wherein the dehydrating
 Pathologists are specialized doctors who
agent in the tissue is removed and replaced
examine the diseased tissues.
with the clearing agent.
HISTOPATHOLOGICAL SPECIMENS  Primary application is to make the tissues
 Histo-Surgical Pathology Specimen transparent.
 Cytopathology Specimen INFILTRATION (IMPREGNATION)
 Autopsy/Post-Mortem Specimen  Process whereby clearing agent is completely
TISSUE PROCESSING removed from the tissue and replaced by a
SOLID STRUCTURES AND TISSUES MUST BE PRESERVED medium that will completely fill all the tissue
AND CAREFULLY PROCESSED IN THE FOLLOWING cavities.
ORDER: Types of Infiltrating / Embedding Media:
1. Fixation 1. Paraffin Wax
2. Decalcification* 2. Celloidin
3. Dehydration 3. Gelatin
4. Clearing 4. Resins
5. Infiltration/ impregnation
EMBEDDING (CASTING / BLOCKING /
6. Embedding
7. Trimming MOLDING)
8. Section- Cutting  Process by which the impregnated tissue is
9. Staining placed into a precisely arranged position in a
10. Mounting mold containing a medium which is then
11. Ringing allowed to solidify.
12. Labeling
TRIMMING
 Done prior to cutting to expose the tissue in
FIXATION the paraffin block.
 Preservation of the tissue by physical or
chemical means
SECTIONING (CUTTING / SECTION-
Goals of Fixation: CUTTING)
1. Primary Aim: Preserve the  Done to produce thin slices of tissues using the
morphologic and chemical integrity of microtome.
the cell in as life like manner. STAINING
2. Secondary Aim: Harden and protect the
 Process of giving color to the tissue
tissue from damage of further handling.
 Most commonly used Staining technique:
DECALCIFICATION Hematoxylin and Eosin Technique
 Special processing technique done to remove MOUNTING
calcium salts from calcified tissues.
 Process of putting a coverslip to the prepared
Methods of decalcification:
sample
3. Use of Acid Decalcifying Agents
4. Use Chelating agents RINGING
5. Use of Ion exchange resins  process of sealing the margins of the cover slip
6. Via Electrophoresis

50
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
LABELING
 process of indicating the year and specimen
number on one end of the prepared slide for
proper identification

 HISTOPATH REPORTS
 Surgical Pathology
 Cytopathology report
 Autopsy report
 SIGNATORIES
 Request Forms
 Result Forms CARE OF THE MICROTOME
***Report ***Retention  After sectioning, all accumulated paraffin and
small pieces of tissues must be brushed away
Autopsy forensic reports Permanently
with a soft brush and not allowed to stay in
Surgical Pathology reports 10 years microtome, since this may later interfere with
Cytogenetics reports 20 years cutting of tissue blocks.
Requisition forms 2 years  Parts should be wiped with xylol/xylene.
 Movable portions should be oiled thoroughly
to prevent rusting.

THE FOLLOWING EQUIPMENT ARE ALSO

REQUIRED DURING THE PROCESS OF
SECTIONING:
 Microtome
 MICROSCOPE
TYPES OF MICROTOME
 WATER BATH- Temperature of the water
- Rocking Microtome
- Rotary Microtome should be about 6 - 10°C below the melting
- Sliding Microtome point of paraffin wax.
- Freezing Microtome  DRYING OVEN- its temperature setting is at the
- Ultra thin Microtome melting point of the wax to allow the sections
to dry.
 ***ROTARY MICROTOME AKA MINOT IS THE  FORCEPS / SABLE HAIRBRUSH- needed for
MOST COMMONLY USED TYPE (PARAFFIN handling sections during cutting, and for
EMBEDDED SECTIONS) removing folds and creases on the sections
 Rocking for large blocks or paraffin during “Floating out” in water bath.
embedded sections  FROSTED END CLEAN GLASS SLIDES-
 Rotary paraffin embedded sections 76x25 mm slides that are 1.0-1.2mm thick are
 Sliding celloidin embedded sections usually preferred because they do not break
 Freezing using cryostat easily.
- ***For cutting frozen sections
 Ultrathin for cutting sections for Electron
 COPLIN JARS / SLOTTED STAINING DISHES
microscopy
51
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
FACTORS INFLUENCING THE IMMUNE RESPONSE:
 Age
 Overall health
 Dose
 Route of inoculation
IMMUNOLOGY/ SEROLOGY SECTION
IMMUNOLOGY- Defined as the study of the

reactions of a host when foreign substances
ANTIBODY
are introduced into the body.  A.K.A. IMMUNOGLOBULINS
 ANTIGEN- these are foreign substances  A gamma globulin protein
that can induce immune response. It can  Produced by “Plasma Cells”
be harmful/ harmless.  They play an essential role during “Antigen
 ANTIBODY- Serum factors in the blood recognition” and in biological activities related
formed in response to foreign to immune response such as “opsonization”
substance exposure. and “complement activation”
 Antibodies are produce by  Classification of immunoglobulins: IgG, IgA,
plasma cells. IgM, IgD and IgE
 SEROLOGY- Study of serum and is known for THERE ARE FIVE MAJOR IMMUNOGLOBULIN CLASSES:
the qualitative detection or quantitative analysis  IgM
of antibodies or antigens concerning infection  IgA
or disease diagnosis.  IgD
 IgG
IMMUNITY  IgE
 All those physiological mechanisms that endow
the animal with the capacity to recognize
materials as foreign to itself and to neutralize,
 The response of the body will depend on the
eliminate or metabolize them with or without
antigen introduced into the host.
injury to its own tissues.
 Factors to consider:
 The immune system is structured to
 First line of defense  Natural
recognize, respond to, and destroy a wide
 Second line of defense  Natural
variety of invading organism that would
 Third line of defense  Acquired
otherwise be capable of promoting infections,
harmful to the body.
TYPES OF IMMUNITY
 NATURAL IMMUNITY: ability of an individual to
resist infections by means of normally present
body functions.  Blood is collected aseptically by
 ACQUIRED IMMUNITY: type of resistance that is venipuncture into a clean, dry, sterile tube.
characterized by specificity for each  Care must be taken to avoid hemolysis, since
individual pathogen, or microbial agent. this may produce a false positive test.
ANTIGEN  Serum should be promptly separated into
another tube without transferring any cellular
 The immune response of lymphocytes is
elements.
triggered by materials called “immunogens”.
 Fresh, non-heat inactivated serum is usually
 The term antigen refers to a substance that
recommended for testing.
reacts with antibody or sensitized T cells but
 There are certain serological test that
may not be able to evoke an immune
requires inactivated serum.
response in the first place.
52
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 Like VDRL and FTA-Abs
 Serum should be heated at
56oC for 30 minutes
 If testing cannot be performed immediately 2-
8oC for up to 72hrs, -20oC if >72 hours delay.

 Depending on the scope of testing performed in

an immunology laboratory, transporting and
receiving patient specimens must be
considered.
 Regulations for packaging and labeling
developed by the U.S. Department of
Transportation (DOT), the International Air
Transport Association (IATA), and the United
Nations must be followed.
CHARACTERISTIC OF A TRANSPORT
CONTAINER SUSPECTED
INFECTIOUS SPECIMEN- DOT AND
IATA RULES
 Watertight primary containers made of
glass, metal, or plastic with a positive (screw-
on) cap.
 The primary container must be wrapped
with enough absorbent material to be
capable of absorbing all of its contents.
Multiple specimens must be wrapped
individually prior to placing them in the
leak-proof secondary container.
 The secondary container is placed in a sturdy
outer container made of corrugated
fiberboard, wood, metal, or rigid plastic. An
itemized list of contents in a sealed plastic bag is
also placed in the outer container.
 Ice packs are placed between the secondary
and the outer container. Additional
measures must be taken when using ice and BLOOD BANK SECTION
dry ice.
 The section of the laboratory where blood may
 In January 2007, labeling of the outer
be collected, stored, and prepared for
container changed.
transfusion.
 The terms clinical specimen and diagnostic
 In the blood bank, blood from patients and
specimen have been replaced with biological
donors is tested for its blood group (ABO)
substances, Category B.
and Rh type.
 This wording is placed next to the label UN
3373.
53
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
Tests performed in Blood Bank Section

 AMERICAN ASSOCIATION OF BLOOD BANKS

(AABB)- was established in 1947. It is an
international association of blood banks that
 The presence and identity of abnormal includes hospital and community blood
antibodies, and its compatibility (crossmatch) centers, transfusion and transplantation
for use in a transfusion. centers, and individuals involved in transfusion
 Units of blood are collected from donors, tested medicine. The mission of AABB is to establish
for the presence of bloodborne pathogens such and provide the highest standard of care for
as hepatitis viruses and human patients and donors in all aspects of
immunodeficiency virus (HIV). transfusion medicine.
 FOOD AND DRUG ADMINISTRATION (FDA) inspects
blood banks on an annual basis; its regulations
for donor are outlined in the Code of Federal
Regulations, parts 211, 600-799.
 Whole Blood
 Packed Red Cells DONOR SCREENING
 Platelet Concentrate  Encompasses the medical history
 Fresh Frozen Plasma requirements for the donor, the physical
 Cryoprecipitate examination, and serologic testing of the
donor blood.
MEDICAL HISTORY AND PHYSICAL EXAMINATION
 Blood bank samples are collected in plain red Is designed to answer two questions:
(serum), lavender, or pink (plasma) stopper  Will a donation of approximately 450 mL of
tubes. whole blood at this time be harmful to the
 Serum separator tube containing gel are not donor?
acceptable because the gel will coat the RBCs  Could blood drawn from this donor at this time
and interfere with testing. potentially transmit a disease to the
 Hemolysis also interferes with the recipient?
interpretation of test results.
MEDICAL HISTORY
 Patient identification is critical in the blood
 Obtaining an accurate medical history of the
bank, and phlebotomists must carefully follow
donor is essential to ensure benefit to the
all patient identification and sample labeling
recipient.
procedures to ensure that a patient does not
 The interviewer should be familiar with the
receive a transfusion with an incompatible
questions, and the interview should be
blood type.
conducted in a secluded area of the blood
center.

54
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
 The questions are designed so that a simple  James Homer Wright (1902) development of
“yes” or “no” can be answered but elaborated Wright stain
if indicated.
 The medical history is conducted on the same HEMATOLOGY
day as the donation.  The study of the formed (cellular) elements
 Medications the donor taking are present in of the blood.
plasma, may cause deferral  Deals with the physiology, pathology,
 Infections the donor has may be passed to etiology, diagnosis, treatment, prognosis and
recipient, may be cause for deferral prevention of blood-related disorders.
PHYSICAL EXAMINATION
 Provides a general screening of health and Formed elements: RBCs, WBCs, Platelets
vital signs to ensure good health on the day of
donation. FORMED ELEMENTS
 Specific screening assessments are performed
and the results are recorded in the donor
record.

 GENERAL APPEARANCE  STEP 1: PHLEBOTOMY

 Observe the prospective donor for  STEP 2: CENTRIFUGATION PROCESS
presence of excessive anxiety, drug or  STEP 3: SEPARATION OF COMPONENTS
alcohol influence, or nervousness.
 This should be done in a gentle manner
so as to not deter the donor from
donations in the future.
 TEMPERATURE- Less than or equal 37.5°C or
99.5°F
 BLOOD PRESSURE- Systole: Less than or equal to
180 mm Hg
Diastole: Less than or equal 100 mm Hg
 PULSE- 50-100 beats per minute
 *HEMOGLOBIN- Greater than or equal to 12.5
g/dL
 *HEMATOCRIT- greater than or equal to 38%
 WEIGHT- 50 kg or 110 lbs.

HISTORY
 Athanasius Kircher (1657) described “worms”
in the blood
 Anton van Leeuwenhoek (1674) gave an
account of RBCs
 Giulio Bizzozero (1800s) described platelets
as “petites plaques”

55
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 Occurs in red bone marrow, or myeloid tissue.

In adults, this tissue is found chiefly in the axial
skeleton.
 Formed elements arise from a common stem
cell, the hemocytoblast (“blood cell former”).
 LYMPHOID STEM CELL
 MYELOID STEM CELL

FORMED ELEMENTS OF THE

BLOOD
ERYTHROCYTES
 Immature Erythrocytes (Reticulocyte)
 Mature Erythrocytes (Discocyte)
LEUKOCYTES
 GRANULOCYTES
- Neutrophils
- Eosinophils
- Basophils
 AGRANULOCYTES
- Lymphocytes
- Monocytes
- Plasma Cells
THROMBOCYTES

 More commonly known as Red Blood Cells.

 Anucleate, biconcave discoid cell that contain
reddish protein that transports gases to and
from different parts of the body.
 RBCs appear pink to red.
 With a zone of pallor that occupies one third
of their center.
 Lifespan: *120 Days

56
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
RBC MEMBRANE  Unsegmented or bilobed
 50% Proteins  release mediators of inflammation
 40% Lipids
 10% Carbohydrates

GRANULOCYTES
 Neutrophils (50-70%)
 Eosinophils (1-3%)
 Basophils (0-2%)
AGRANULOCYTES
 Lymphocytes (18-42%)
 Monocytes (2-11%)
LYMPHOCYTE
 Pale to moderate blue cytoplasm
NEUTROPHIL  Round or oval-shaped nucleus and may be
 Has pink to rose-violet specific granules slightly indented.
 Multilobed (2-5 lobes) nuclei  Slightly larger than RBCs
 Phagocytic cells  Function in destroying cancer cells, cells
 Engulfs and destroys microorganisms and infected by viruses, and foreign invading cells.
foreign material

MONOCYTE
EOSINOPHIL  Blue-gray cytoplasm
 Has reddish-orange granules
 Contains fine azure granules
 Usually has 2 lobes
 Round, kidney shaped nucleus and may show
 Release enzymes that disable parasites brain-like convolutions
 Phagocytic cells in many places of body

BASOPHIL
 Has dark-purple to blue-black irregular
granules
57
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

 More commonly known as platelets

 Measure 2-4um in diameter
 Light blue to purple, very granular
 Responsible for blood clotting

THICK BLOOD SMEAR

 Used to screen for blood parasitic organisms
THIN BLOOD SMEAR
 Used to identify blood parasites
 Used in visualizing and identifying blood
cells (manual differential counting)
WHOLE BLOOD
 The most common body fluid analyzed in the
hematology section.
 Obtained by using a collection tube with an
anticoagulant.
*Most tests performed in the hematology section
require blood that has been collected in tubes with a
lavender stopper that contain the anticoagulant EDTA
CELL COUNTING
 Manual Cell Count WEDGE TYPE SMEAR
 Improved Neubauer Counting
Chamber (Hemocytometer)
 Automated Cell Count

NEUBAUER COUNTING CHAMBER

58
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA
THIN BLOOD FILM
PREPARATION:
 Transfer a drop of blood to the Stationary slide
about .25 inch from the frosted end
 Position the spreader slide at a 25<, allow the
blood to spread to the back edge of the spreader
slide.
 Immediately push the spreader slide forward
with a smooth and rapid stroke maintaining the
angle
 Air dry rapidly but thoroughly before staining

QUALITIES OF A GOOD THIN

BLOOD FILM:
 Cover at least half the length of the slide
 Smear should be narrower than the slide
 Smooth spread should display a gradual
transition from thick to thin
 No waves
 No streaks
 No troughs
 No holes or bubbles
 Straight feathered edge
 Contain at least 10 LPF in which 50% of the
RBCs do not overlap

Factor Thick smear Thin smear PROCEDURE

Increased  Allow smears to dry
Pressure Light Pressure  Dip slide five times, for one second each, into
pressure
Fixative. Allow excess to drain after each dip.
High angle of  Dip slide five times, for one second each, into
Low angle of the Stain 1. Allow excess to drain after each dip.
Angle the spreader
spreader slide  Dip slide five times, for one second each, into
slide
Stain 2. Allow excess to drain after each dip.
Size of  Rinse slide or tape-strip in distilled water or
the Large drop of Small drop of Weise's buffer, pH 7.2.
blood blood blood  Blot or allow to dry in air.
drop  Examine at low power to identify structures and
then under oil immersion.
Speed Fast spreading Slow spreading

59
MLSP 112_1ST YEAR_2ND SEM ©ABEGAIL G MAGBANUA

END OF SEMESTER

END OF SEMESTER 60