PHLEB LAB Different Methods of Handwashing:

Module 1: Laboratory Safety and

1. A routine hand-washing procedure
Personal Hygiene
Mikee Patricia D. Trinidad ꟾ BSMLS1-G
used plain soap and water to
mechanically remove soil and transient
bacteria.
2. Hand antisepsis requires the use of an
antimicrobial soap to remove, kill, or
Hand Hygiene
inhibit transient microorganisms.
Hand hygiene is one of the most important 3. A 2-minute surgical hand scrub uses
means of preventing the spread of infection an antimicrobial soap or equivalent to
provided that it is achieved properly and when remove or destroy transient
required. Hand hygiene measures include the microorganisms and reduce levels of
frequent use of antiseptic hand cleaners or normal flora prior to surgical
hand washing, depending upon the degree of procedures.
contamination. It is important that all
Hand-Washing Technique Steps:
healthcare personnel learn proper hand
hygiene procedures and recognize situations Step 1: Stand back so that you cannot touch
when they should be performed. the sink
Here is the list of situations that require hand Rationale: The sink may be contaminated
hygiene procedures:
• Before and after each patient contact.
• Between unrelated procedures on a Step 2: Turn on the faucet and wet hands
patient such as wound care and under warm running water
drawing blood. Rationale: Water should not be too hot nor
• Before putting on gloves and after taking too cold and hands should be wet before
them off. applying soap to minimize drying,
• Before leaving the laboratory. chapping, or cracking of hands from
• Before going to lunch or on break. frequent hand washing.
• Before and after going to the restroom.
• Whenever hands become visibly or
knowingly contaminated. Step 3: Apply soap and work up a lather
As part of the CDC Guidelines for Hand Hygiene Rationale: A good lather is needed to reach
in Health Care Settings, it is recommended that all surfaces
artificial fingernails or extenders not be worn
when having direct contact with high-risk
patients, such as infants or those in ICU. Step 4: Scrub all surfaces, including
Natural nails should be preferably less than ¼ between the fingers and around the
inch long. knuckles

If hands are heavily contaminated with organic Rationale: Scrubbing is necessary to

matter and hand-washing facilities are not dislodge microorganisms from surfaces,
available, it is recommended that hands be especially between fingers and around
cleaned with detergent-containing wipes, knuckles.
followed by the use of an alcohol-based
antiseptic hand cleaner.
Step 5: Rub your hands together vigorously
Rationale: Friction helps loosen dead skin,
dirt, debris, and microorganisms
 of properly, it prevents the spread of infection
to others.
Steps 4 and 5 should take at least 15
seconds, about the time it takes to sing the 1. Gloves
ABCs
Gloves are made of a variety of materials (e.g.,
vinyl, nitrile, latex) and come in a variety of
sizes. Latex used to be the most common
Step 6: Rinse your hands in a downward
material for gloves, but because of the increase
motion from wrists to fingertips
in latex allergies, most health care facilities are
Rationale: Rinsing with hands downward phasing out their use of latex gloves and
allows contaminants to be flushed from the replacing them with gloves made of either vinyl
hands and gingers into the sink rather than or nitrile.
flowing back up the arm or wrist
Clean, sterile gloves are worn when collecting
or handling blood and other body fluids,
handling contaminated items, and touching
Step 7: Dry hands with a clean paper towel nonintact skin or mucous membranes.
Rationale: Hands must be dried thoroughly Three (3) Main Reasons for Wearing Gloves
and gently to prevent chapping or cracking.
Reusable towels can be a source of • To prevent contamination of the hands
contamination when handling blood or body fluids or
when touching mucous membranes or
nonintact skin
Step 8: Use a clean paper towel to turn off • To reduce the chance of transmitting
the faucet unless it is foot or motion- organisms on the hands of personnel to
activated patients during invasive or other
procedures that involve touching a
Rationale: Clean hands should not touch patient’s skin or mucous membrane
contaminated faucet handles • To minimize the possibility of
transmitting infectious microorganisms
from one patient to another
7 Steps of Handwashing with Soap and Water
1. Palm to palm
2. Between fingers Proper Glove Removal
3. Back of hands 1. Pinch and hold the outside area of the
4. Base of thumbs glove near the wrist area
5. Back of fingers 2. Peel downwards, away from the wrist,
6. Fingernails turning the glove inside-out
7. Wrists 3. Pull the glove away until it is removed
8. Rinse and wipe dry (7 lang pro ngaa 8 sa from the hand, holding the inside-out
module tf) glove with the gloved hand
4. With the un-gloved hand, slide the
finger/s under the wrist of the remaining
Personal Protective Equipment glove. Do not touch the outer surface of
Protective clothing and other items worn by an the glove
individual to protect mucous membranes, 5. Peel downwards, away from the wrist,
airways, skin, and clothing from contact with turning the glove inside out
infectious substances are called personal 6. Continue to pull the glove down and
protective equipment (PPE). over the inside-out glove being held in
your gloved hand
PPE provides a barrier against infection. Used
properly, it protects those wearing it. Disposed
 2. Gowns 5. Respirators
Fluid-resistant gowns provide full-body NIOSH-approved N95 respirators are required
coverage and prevent body fluids or spills from when entering rooms of patients with
passing through and contacting the skin or pulmonary tuberculosis and other diseases
clothing. The cuffs are designed to close tightly with airborne transmission. Respirators must
around the wrists and are covered by the fit snugly with no air leaks.
gloves for further protection.
Donning and Doffing of PPE
Alternatively, a cloth gown may be worn to
Donning
prevent contamination of skin and clothing
with transmissible infectious microorganisms 1. Hand hygiene
when protection from fluid penetration is not 2. Gown
necessary. 3. Mask
4. Goggles
5. Gloves
3. Lab Coats
Lab coats, like gowns, are worn to protect skin
Donning
and prevent soiling of healthcare workers'
clothing during patient-care activities in which 1. Gloves
splashes, or sprays of blood or body fluids are 2. Gown
possible. 3. Goggles
4. Mask
They are required attire for most phlebotomy
5. Hygiene
situations. Lab coats used for specimen
collection and handling are generally made of
fluid-resistant cotton or synthetic material,
have long sleeves with knit cuffs, and come in Occupational Safety and Health
both reusable and disposable styles. Administration
Workplace safety is regulated by OSHA. The
regulations are designed both to inform
4. Masks, Face Shields, and Goggles workers about hazards in the workplace (e.g.,
by requiring that workers know the health
A mask is worn to protect against droplets
effects of the chemicals they use) and to
generated by coughing or sneezing. To put on a
protect workers from harm (e.g., by requiring
mask, place it over your nose and mouth.
an emergency shower nearby in case of
Adjust the metal band (if applicable) to fit
chemical spills).
snugly over your nose. For masks with ties,
fasten the top ties around the upper portion of Your employer is required by OSHA to maintain
your head; then tie the lower ones at the back a safe workplace, provide a comprehensive
of your neck. If the mask has elastic fasteners, safety training program, and report accidents
slip them around your ears. that occur on the job. OSHA regulations are
revised as needed to increase workplace safety
A face shield or a mask and goggles are worn
in light of new information or new hazards.
to protect the eyes, nose, and mouth from
Therefore, you should keep up to date on all
splashes or sprays of body fluids. If an activity
relevant information as it changes throughout
requires goggles, it also requires a mask. Some
your career.
masks have plastic eye shields attached.
Types of Hazards
1. Biological
Infectious agents, including airborne or
bloodborne organisms such as bacteria and
viruses

2. Physical
Wet floors, heavy lifting (e.g., boxes, patient
transfers)
Avoiding physical hazards in the workplace is 3. Sharps
mostly a matter of common sense, plus
learning some important habits: Needles, lancets, and broken glass

• Avoid running. This is not only a safety Sharps, especially needles and lancets, are the
rule but also a consideration for patients most common hazards you will encounter as a
who may become concerned or phlebotomist. Sharps are dangerous both
agitated because of the physical injury they may cause
• Watch for wet floors. Your facility should and because they may carry bloodborne
use cones or other types of signs to warn pathogens such as human immunodeficiency
when a floor is wet and should have virus (HIV), hepatitis B virus (HBV), or hepatitis
cleanup equipment for spills C virus (HCV).
• Bend your knees when lifting heavy
objects or transferring a patient
• Maintain a clean, organized work area
 4. Chemical 6. Electrical
Preservatives and reagents (laboratory-grade Dangerous high-voltage equipment
chemicals)
Electrical hazards in the laboratory may result
You will encounter many different chemicals in in shock or fire
your work as a phlebotomist, including several
General rules of electrical safety:
that can be quite harmful if handled
improperly. • Avoid using extension cords.
• Report frayed cords, overloaded
For example, hydrochloric acid (HCI), which
circuits, and ungrounded equipment.
burns mucosal tissue and skin, is used as a
• Unplug a piece of equipment before
preservative for urine. Bleach, which causes
servicing it.
irritation of mucosal tissue and skin, is used as
• If a piece of equipment is marked with
a disinfectant in the laboratory.
an electrical caution warning, do not
attempt to open it, even for inspection.
It may contain batteries or electrical
capacitors that store electricity even
when unplugged.
• Know the location of the circuit breaker
box for the equipment you are using.
• Avoid contact with any electrical
equipment while drawing blood.
Electricity may pass through you and
the needle and shock the patient.

5. Radiation
Radioactive reagents, X-ray equipment
The principle involved in radiation exposure are
distance, shielding, and time.
This means that the amount of radiation you
are exposed to depends upon how far you are
from the source of radioactivity, what
protection you have from it, and how long you 7. Fire or explosive
are exposed to it. Open flames, oxygen, chemicals (e.g., nitrous
Exposure time is important because radiation oxide), and Gases under pressure
effects are cumulative. Fires or explosions in the laboratory may occur
as a result of chemical or electrical accidents or
carelessness with flames or other fire sources.
In addition to preventive measures, the most
important steps to take to minimize the risk of
injury are summarized in the acronym RACE:
R: Rescue
A: Alarm
C: Contain Fire
E: Extinguish/evacuate
PASS A second type of skin allergy called "allergic
contact dermatitis" may be caused by
P: Pull the pin
chemicals used to manufacture rubber gloves.
A: Aim at the base This dermatitis is recognized by eczema and
blisters on the back of the hands. It resembles
S: Squeeze trigger a poison ivy rash and begins 1 to 3 days after
S: Sweep wearing rubber gloves.
Direct physical contact with latex products is
not needed to trigger an allergic reaction.
Anaphylaxis and severe asthmatic reactions
have been caused by inhaling latex proteins in
the air resulting from the powder in the latex
glove.

8. Latex sensitivity
Allergic reaction to latex in gloves or other
equipment
In most cases, latex allergy develops after
many previous exposures to latex. Latex
allergy symptoms may include hives, itching,
stuffy or runny nose. It can cause asthma
symptoms of wheezing, chest tightness, and
difficulty breathing.
Symptoms begin within minutes after exposure
to latex-containing products. The most severe
latex allergy can result in anaphylaxis, a
serious allergic reaction involving severe
breathing difficulty and/or fall in blood pressure
(shock).
Allergic skin problems can occur following
direct contact with allergic latex proteins in
latex glove products. Symptoms may include
immediate itching, redness, and swelling of the
skin that touched the item containing latex.
These and other latex allergic reactions are less
common now. Many hospitals or doctors'
offices have switched to non-latex gloves or low
protein latex gloves.
 PHLEB LAB • Left atrium contracts, the oxygenated
Module 2: The Circulatory System blood goes to the left ventricle
Mikee Patricia D. Trinidad ꟾ BSMLS1-G • Left ventricle contracts, the oxygenated
blood is pumped into the aorta
Aorta is the largest artery in the body, and its
function is to supply the body with oxygenated
The circulatory system carries oxygen and food blood.
to the cells of the body and carries carbon
dioxide and other wastes away from the cells
to the excretory organs, kidneys, lungs, and The Systemic Circulation
skin. The systemic circulation serves the rest of the
It also aids in the coagulation process, assists body, carrying oxygenated blood and nutrients
in defending the body against disease, and from the left ventricle of the heart to the body
plays an important part to the phlebotomist, cells and then returning to the right atrium of
who must access it to collect blood specimens the heart with blood carrying carbon dioxide
for analysis. It also helps the phlebotomist and other waste products of metabolism from
appreciate the importance of the many tests the cells.
associated with it.

STRUCTURES
The Vascular System The structures of the vascular system are the
FUNCTIONS various blood vessels that, along with the heart,
form the closed-system through which blood
The vascular system is the system of blood flows.
vessels that, along with the heart, forms the
closed loop through which blood is circulated to Blood vessels are tubelike structures capable of
all parts of the body. There are two divisions to expanding and contracting. According to
this system, pulmonary circulation, and information from the Arizona Science Center,
systemic circulation. the human vascular system has around
250,000 miles of blood vessels, 95% of which
are capillaries, which make up what is called
The Pulmonary Circulation the capillary bed. The rest are arteries and
veins.
The pulmonary circulation carries blood from
the right ventricle of the heart to the lungs to
remove carbon dioxide and pick up oxygen; the Arteries
oxygenated blood is then returned to the left
atrium of the heart. Arteries are blood vessels that carry blood
away from the heart. They have thick walls
• Right atrium receives deoxygenated because the blood that moves through them is
blood from the superior and inferior under pressure from the contraction of the
vena cava ventricles.
• Right atrium contracts, the
deoxygenated blood goes to the right This pressure creates a pulse that can be felt,
ventricle distinguishing the arteries from the veins.
• Right ventricle contracts, deoxygenated NOTE: When arterial blood is collected by
blood is pumped into the lungs through syringe, the pressure normally causes the
the pulmonary artery blood to "pump" or pulse into the syringe under
• Deoxygenated blood is oxygenated in its own power.
the lungs
• Oxygenated blood is carried by the Systemic arteries carry oxygenated (oxygen-
pulmonary vein to the left atrium rich) blood away from the heart to the tissues.
Because it is oxygen-rich or full of oxygen, Blood in the capillaries is a mixture of both
normal systemic arterial blood is bright red. venous and arterial blood. In the systemic
circulation, arterial blood delivers oxygen and
The pulmonary artery is the only artery that
nutrients to the capillaries. The thin capillary
carries deoxygenated, or oxygen-poor, blood.
walls allow the exchange of oxygen for carbon
It is part of the pulmonary circulation and
dioxide and nutrients for wastes between the
carries deoxygenated blood from the heart to
cells and the blood.
the lungs. It is classified as an artery because
it carries blood away from the heart. Carbon dioxide and wastes are carried away
in the venous blood. In the pulmonary
The smallest branches of arteries that join with
circulation, carbon dioxide is delivered to the
the capillaries are called arterioles.
capillaries in the lungs and exchanged for
The largest artery in the body is the aorta. It is oxygen.
approximately 1 inch (2.5 cm) in diameter.

Veins
Veins are blood vessels that return blood to the
heart.
Veins carry blood that is low in oxygen
(deoxygenated or oxygen-poor) except for the
pulmonary vein, which carries oxygenated
blood from the lungs back to the heart.
Because systemic venous blood is oxygen-
poor, it is much darker and more bluish-red
than normal arterial blood.
The walls of veins are thinner than those of BLOOD VESSEL STRUCTURE
arteries because the blood is under less Arteries and veins are composed of three main
pressure than arterial blood. Since the walls are layers. The thickness of the layers varies with
thinner, veins can collapse more easily than the size and type of blood vessel. Capillaries
arteries. are composed of a single layer of endothelial
Blood is kept moving through veins by skeletal cells enclosed in a basement membrane.
muscle movement, valves that prevent the
backflow of blood, and pressure changes in the
abdominal and thoracic cavities during Layers:
breathing. Tunica adventitia: the outer layer of a blood
The smallest veins at the junction of the vessel, sometimes called the tunica externa. It
capillaries are called venules. The largest veins is made up of connective tissue and is thicker
in the body are the venae cavae (singular, vena in arteries than in veins.
cava). The longest veins in the body are the Tunica media: the middle layer of a blood
great saphenous veins in the leg. vessel. It is made up of smooth muscle tissue
and some elastic fibers. It is much thicker in
arteries than in veins.
Capillaries
Tunica intima: the inner layer or lining of a
Capillaries are microscopic, one-cell-thick blood vessel, sometimes called the tunica
vessels that connect the arterioles and venules, interna. It is made up of a single layer of
forming a bridge between the arterial and endothelial cells with an underlying basement
venous circulation. membrane, a connective tissue layer, and an
elastic internal membrane. It has direct up in the process of clot formation. Some
contact with blood. chemistry tests, such as ammonia and
potassium, cannot be performed on serum
because clotting releases these substances
Valves from the cells.
Venous valves are thin membranous leaflets In addition, some chemistry test results are
composed primarily of epithelium similar to needed stat (immediately) in order to respond
that of the semilunar valves of the heart. Most to emergency situations; having to wait 30
of the venous system flows against the pull of minutes or more for a specimen to clot before
gravity. centrifuging it to get serum would be
unacceptable.
As blood is moved forward by the movement of
skeletal muscle, for example, the valves help If clotting is prevented, however, coagulation
keep it flowing toward the heart by allowing factors and other substances affected by
blood to flow in only one direction. clotting are preserved, and the specimen can
be centrifuged immediately. Blood can be
Contracting skeletal muscle compresses the prevented from clotting by adding a substance
vein and drives blood forward, opening the called an anticoagulant. Adding an
proximal valve, while the distal valve closes to anticoagulant initially creates a whole-blood
prevent the backflow of blood. When the specimen.
muscles relax again, the distal valve opens and
the proximal valve closes until blood moving in When a whole-blood specimen is centrifuged,
the vein forces it to open again. it separates into three distinct layers: a bottom
layer of red blood cells; a thin, fluffy-looking,
whitish-colored middle layer of WBCS and
Blood Specimens platelets referred to as the buffy coat; and a top
layer of liquid called plasma, which can be
Serum separated from the cells and used for testing.
Blood that has been removed from the body Normal fasting plasma is a clear to slightly
will coagulate or clot within 30 to 60 minutes. hazy pale-yellow fluid visually indistinguishable
The clot consists of the blood cells enmeshed in from serum. The major difference between
a fibrin network. plasma and serum is that plasma contains
The remaining fluid portion is called serum and fibrinogen. Many laboratory tests can be
can be separated from the clot by performed on either serum or plasma.
centrifugation (spinning the clotted blood at a
very high speed in a machine called a
centrifuge). Whole Blood

Normal fasting serum is a clear, pale yellow Some tests, including most hematology tests
fluid. Serum has the same composition as and some chemistry tests such as
plasma except that it does not contain glycohemoglobin, cannot be performed on
fibrinogen because the fibrinogen was used in serum or plasma. These tests must be
the formation of the clot. Many laboratory performed on whole blood (blood in the same
tests, especially chemistry and immunology form as it is in the bloodstream). This means
tests, are performed on serum. that the blood specimen must not be allowed
to clot or separate.
To obtain a whole-blood specimen, it is
Plasma necessary to add an anticoagulant. In addition,
Not all tests can be performed on serum. For because the components will separate if the
example, most coagulation tests cannot be specimen is allowed to stand undisturbed, the
performed on serum because some of the specimen must be mixed for a minimum of 2
coagulation factors (e.g., fibrinogen) are used
minutes immediately prior to performing the C. Secondary Hemostasis Plug Formation
test.
For injuries with more tissue damage, the
stimulus for coagulation is strong, and the
coagulation process continues until the initial
Basic Hemostasis and Fibrinolytic Process
platelet plug becomes a stable blood clot called
Hemostasis, which means the arrest or the secondary hemostatic plug.
stoppage of bleeding, is the body response that
This involves the amplification and propagation
stops the loss of blood after injury without
phases of the coagulation process.
affecting the flow of blood within the rest of the
vascular system. (The opposite of hemostasis is
hemorrhage.) The hemostatic process requires
D. Fibrinolysis
the coordinated interaction of endothelial cells
lining the blood vessels, platelets, other blood Fibrinolysis is the process by which fibrin is
cells, plasma proteins, and the coagulation dissolved. It is an ongoing process responsible
(clotting) process. for two important activities: (a) it dissolves
clots that form within intact vessels (thrombi),
This involves four interrelated responses: (a)
thus reopening the vessels, and (b) it removes
vasoconstriction; (b) formation of a primary
hemostatic clots from the tissue as healing
platelet plug in the injured area; (c)
occurs.
progression, if needed, to a stable blood clot
called the secondary hemostatic plug; and (d) The process is possible because activation of
fibrinolysis, the dissolving of the clot after the the clotting process also activates factors like
site has healed. Xlla and promotes the release of plasminogen
activators from vessel lining cells and white
blood cells.
A. Vasoconstriction
These substances convert plasminogen to
The immediate reaction to blood vessel injury plasmin. Plasmin is the enzyme that breaks
is vasoconstriction, a reduction in the diameter down fibrin into smaller fragments called fibrin
of the blood vessel caused by the contraction of degradation products, which are then removed
smooth muscle fibers in the tunica media. This by phagocytic cells.
narrows the blood vessel, which decreases the
flow of blood past the injured area and limits
blood loss. Fibrin- an insoluble protein formed from
fibrinogen during the clotting of blood. It forms
Vasodilation prevents the vessel from
a fibrous mesh that impedes the flow of blood
becoming too narrow and blocking effective
blood flow. Fibrinogen- a soluble protein present in blood
plasma, from which fibrin is produced by the
action of the enzyme thrombin
B. Primary Platelet Plug Formation
Platelet plug formation occurs during the
initiation phase of the coagulation process; it
involves the adhesion, activation, and
aggregation of platelets and the generation of
a small amount of fibrin. Normal platelet plug
formation depends on an adequate
concentration of platelets (determined by a
platelet count), normally functioning platelets
(determined by platelet aggregation testing),
and blood vessel integrity.
Major Veins for Venipuncture possibility of accidental puncture of the
anterior or posterior branch of the
Antecubital Fossa
medial cutaneous nerve (a major nerve
Antecubital means "in front of the elbow." of the arm) or the brachial artery, both
Fossa means a shallow depression. The of which commonly underlie this area.
antecubital (AC) fossa is the shallow Punctures in this area also tend to be
depression in the arm that is anterior to (in more painful.
front of) and below the bend of the elbow. It is
the first-choice location for venipuncture
because several major arm veins lie close to
the surface in this area, making them relatively
easy to locate and penetrate with a needle.
These major superficial veins are referred to as
antecubital veins. The anatomical
arrangement of antecubital veins varies
slightly from person to person; however, two
basic vein arrangements, referred to as the H-
and M-shaped patterns are seen most often.

H-Shaped Antecubital Veins

The H-shaped venous distribution pattern is
displayed by approximately 70% of the
population and includes the median cubital
vein, cephalic vein, and basilic vein. M-Shaped Antecubital Veins

• Median cubital vein: Located near the The veins that form the M-shaped venous
center of the antecubital area, it is the distribution pattern include the cephalic vein,
most preferred vein for venipuncture in median vein, median cephalic vein, median
the H-shaped pattern. It is typically basilic vein, and basilic vein.
larger, closer to the surface, better The veins most commonly used for
anchored, and more stationary than the venipuncture in this distribution pattern are
others, making it the easiest and least described as follows:
painful to puncture and the least likely
to bruise. • Median vein (also called the
intermediate antebrachial vein): The
• Cephalic vein: Located in the lateral first choice for venipuncture in the M-
aspect of the antecubital area, it is the shaped pattern because it is well
second-choice vein for venipuncture in anchored, tends to be less painful to
the H-shaped pattern. It is often harder puncture, and is not as close to major
to palpate than the median cubital but nerves or arteries as the others, making
is fairly well anchored and often the only it generally the safest one to use.
vein that can be palpated (felt) in obese
patients. • Median cephalic vein (also called the
intermediate cephalic vein): The second
• Basilic vein: A large vein located on the choice for venipuncture in the M-
medial aspect (inner side) of the shaped pattern because it is accessible
antecubital area, it is the last-choice and is for the most part located away
vein for venipuncture in either venous from major nerves or arteries, making it
distribution pattern. It is generally easy generally safe to puncture. It is also less
to palpate but is not as well anchored likely to roll and relatively less painful to
and rolls more easily, increasing the puncture.
 • Median basilic vein (also called the
intermediate basilic vein): The last
choice for venipuncture in the M-
shaped pattern (even though it may
appear more accessible) because it is
more painful to puncture and, like the
basilic vein, is located near the anterior
and posterior branches of the medial
cutaneous nerve and the brachial
artery.

Other Arm and Hand Veins

According to CLSI, although the larger and
fuller median, median cubital, cephalic, and
median cephalic veins are used most
frequently, veins on the back of the hand and
wrist are also acceptable for venipuncture.
Leg, Ankle, and Foot Veins
Because of the potential for significant medical
complications such as phlebitis or thrombosis,
veins of the leg, ankle, and foot must not be
used for venipuncture without permission from
the patient's physician.
Puncture of the femoral vein is performed only
by physicians or specially trained personnel.
 PHLEB LAB o Benzalkonium chloride (e.g.,
Module 3: Blood Collection Zephiran chloride)
Equipment, Additives, and Order of o Chlorhexidine gluconate
Draw o Hydrogen peroxide
Mikee Patricia D. Trinidad ꟾ BSMLS1-G o Povidone-iodine (0.1%-1%
available iodine)
o Tincture of iodine

General Blood Collection Equipment and • Disinfectants – are chemical solutions

Supplies or substances that are used to remove
or kill microorganisms on surfaces or
• Blood drawing station - A dedicated instruments.
area of a medical laboratory or clinic o 1:100 dilution Sodium
equipped for performing phlebotomy Hypochlorite (household bleach)
procedures on patients. o 1:10 for spills of large amounts
o Includes tables for supplies, a
chair for the patient, and a bed or
• Gauze pads/Cotton pads – Clean 2 by
reclining chair for patients with a
2-inch gauze pads are used to hold
history of fainting, persons pressure over the site following blood
donating blood, and other collection procedures.
situations.
o A bed or padded table for infants • Bandages – Adhesive bandages are
and small children. used to cover a blood collection site
after the bleeding has stopped.

• Equipment Carriers • Needles and Sharps Disposal

o Handheld carriers - trays or containers- special containers to which
carriers that come in various used needles, lancets, and other sharp
sizes and shapes designated to objects are disposed of. It should be
be easily carried by the rigid, puncture-resistant, leakproof, and
phlebotomist and to contain disposable with locking lids. The
enough equipment for numerous container should also be marked with a
draws. biohazard symbol.

o Phlebotomy carts - have several • Tourniquet - is a device applied or tied

shelves to carry adequate around a patient's arm prior to
supplies for obtaining blood venipuncture to restrict the blood flow to
specimens for many patients allow distention or inflation of veins,
making them easier to find.
• Gloves and Glove liners – new pair
must be used for each patient and be • Needles – Phlebotomy needles are
removed when a procedure is complete. sterile, disposable, and designed for
single use only
• Antiseptics – are substances used to o Multisample needles-
prevent sepsis. Antiseptics prevent or evacuated tube system
inhibit the growth and development of o Hypodermic needles- syringe
microorganisms and do not necessarily system
kill them. o Winged infusion (butterfly
Types of Antiseptics used in Blood Collection needles)- used both in the
evacuated tube system and the
o 70% Ethyl alcohol syringe system
o 70% Isopropyl alcohol
(isopropanol)
 Parts of a needle
o Bevel- end that pierces the vein. Gauge: 22
Allows the needle to easily slip Needle Type: Multisample, Hypodermic
into the skin and vein without Typical Use: Used on older children and adult
coring (removing a portion of the patients with small veins or for syringe draws
skin or vein) on difficult veins.
o Shaft- long cylindrical portion
o Hub- end that attaches to the Gauge: 23
blood collection device Needle Type: Butterfly
o Lumen- internal space of the Typical Use: Used on the veins of infants and
needle children or difficult hand veins of adults.

Needle color codes:

• Yellow- 20-gauge
• Green- 21-gauge
• Black- 22-gauge
Gauge- needle
gauge is Needle length
indicated by a number that is related to • Multisample needles- 1 or 1.5-inches
the diameter of the lumen. A needle’s length
diameter and gauge have an inverse • Syringe needles- 1 and 1.5- inches are
relationship; the higher the gauge the ones most commonly used
number, the smaller the actual • Butterfly needles- 1/2 to 3/4 inches long
diameter of the needle. Smaller gauge,
larger diameter.
Evacuated tube system (ETS)
Gauge: 15-17
Needle Type: Special needle attached to a It is a closed system in which the patient's blood
collection bag flows through a needle inserted into a vein and
Typical Use: collection of donor units, then directly into a collection tube without
autologous blood donation, and therapeutic being exposed to the air or outside
phlebotomy contaminants.
1. Multisample Needles- allow multiple
Gauge: 18
tubes of blood to be collected during a
Needle Type: Hypodermic
single venipuncture. They are threaded
Typical Use: used primarily as a transfer
in the middle and have a beveled point
needle rather than for blood collection; safety
from each end. The threaded portion
issues have diminished use
screws into a tube holder. The end of the
needle that pierces the vein is longer
Gauge: 20
and has a longer bevel. The shorter end
Needle Type: Multisample, Hypodermic
penetrates the tube stopper during
Typical Use: sometimes used when large-
specimen collection. It is covered by a
volume tubes are collected or large-volume
sleeve that retracts as the needle goes
syringes are used on patients with normal-size
through the tube stopper so that blood
veins
can flow into the tube. When the tube is
removed, the sleeve slides back over the
Gauge: 21
needle to prevent leakage of blood.
Needle Type: Multisample, Hypodermic
2. Tube holders- a clear, plastic,
Typical Use: needle for routine venipuncture
disposable cylinder with a small
on patients with normal veins or syringe blood
threaded opening at one end, also
culture collection.
called a hub, where the needle is
 screwed into it and a large opening at a. Chelating (binding) or precipitating
the other end where the collection tube calcium so it is unavailable for the
is placed. The large end has flanges or coagulation process
extensions on the sides that aid in tube b. Inhibiting the formation of thrombin
placement and removal. needed to convert fibrinogen to
fibrin
Chelation- binding or removing metals from
the blood

Different Types of Anticoagulants

EDTA (Ethylene Diamine Tetraacetic acid)
• Removes calcium by chelation,
preventing it from ionizing
• Two forms:
o Versene- di-sodium salt of EDTA
o Sequestrene- di-potassium salt
of EDTA has better stability than
Versene
Evacuated tubes
are used with both the ETS and the syringe Citrate
method of obtaining blood specimens. (With
• Prevents coagulation by binding or
the syringe method, blood is collected in a
chelating calcium
syringe and must be immediately transferred
• Intended for blood transfusion because
into the tubes.)
citrate reserves well the cells
• Most common is sodium citrate which
Evacuated tubes are available from a number
is used for coagulation tests since it
of different manufacturers and come in various
preserves the coagulation factors
sizes and volumes ranging from 1.8 to 15 mL.
Heparin

Vacuum - artificially created by pulling • Prevents clotting by inhibiting thrombin

air from the tube. The amount of vacuum, formation
which is the amount of air removed and • Thrombin is an enzyme needed to
negative pressure created, is measured convert fibrinogen to fibrin necessary for
precisely by the manufacturer so that the tube clot formation
will draw the exact volume of blood indicated Oxalates
on the label.
• Prevent coagulation by precipitating
Additives - is any substance placed calcium
within a tube other than the tube stopper. • Potassium oxalate is most widely used
Additives have one or more specific functions, and is commonly added to tubes
such as preventing clotting or preserving containing glucose preservatives
certain blood components, or activating clots
to obtain serum.
Special-use Anticoagulants

Different Types of Additives: Acid Citrate Dextrose

Anticoagulants- are substances that prevent • The acid citrate prevents coagulation by
blood clotting by either of two methods: binding calcium, with little effect on cells
and platelets
 • Dextrose acts as an RBC nutrient and • Tubes containing clot activators require
preservative by maintaining RBC five gentle inversions for complete and
viability rapid clotting to occur.
• ACD tubes have yellow tops and require
Thixotropic Gel Separator
eight inversions immediately after
collection to prevent clotting • Is an inert (nonreacting) synthetic
substance initially contained in or near
Citrate Phosphate Dextrose
the bottom of certain blood collection
• CPD is used in collecting units of blood tubes.
for transfusion. Citrate prevents clotting • The density of the gel is between that of
by chelating calcium, phosphate the cells and the serum or plasma.
stabilizes pH, and dextrose provides • Stoppers or tubes with mottled red/gray
cells with energy and helps keep them rubber stoppers called serum-
alive. separator tubes (SSTS)
• Tubes containing thrombin that clot in 5
Sodium Polyanethol Sulfonate (SPS)
minutes called Rapid Serum Tubes
• SPS prevents coagulation by binding (RSTS)
calcium. It is used for blood culture • Heparinized plasma gel-barrier tubes
collection because it reduces the action include BD tubes with light-green plastic
of a protein called complement which or mottled gray/green rubber stoppers
destroys bacteria. called plasma-separator tubes (PSTS)
• It also slows down phagocytosis • EDTA gel-barrier tubes with pearl-
(ingestion of bacteria by leukocytes), colored stoppers called plasma-
and reduces the activity of certain preparation tubes (PPTS).
antibiotics.
• SPS tubes have yellow stoppers and
require eight inversions to prevent Nonadditive tubes - are additive-free tubes
clotting. that yield serum samples.
Antiglycolytic Agents Stoppers - tube stoppers (tops or closures) are
typically made of rubber that serves to plug
• Is a substance that prevents glycolysis.
joints or holes. Some rubber stoppers have a
• If glycolysis is not prevented, the glucose
plastic shield for extra protection for the
concentration in a blood specimen
aerosols and sprays of the specimen if the
decreases at a rate of 10 mg/dL per
stopper is removed.
hour.
• Most common antiglycolytic agent is Tube stoppers are also color-coded to identify
sodium fluoride and have gray tubes a type of additive placed in the tube.
and require five and ten inversions
Clot activators
SST- serum seperator tubes
• A clot activator is a substance that
PST- plasma separator tubes
enhances coagulation in tubes used to
collect serum specimens.
• Clot activators include substances that
provide more surface for platelet
activation, such as glass (silica)
particles and inert clays like Celite, and
clotting factors such as thrombin. Silica
particles are the clot activators in
serum-separator tubes (SSTS) and
plastic red-top tubes.
Order of Draw
• A special sequence of tube collection
that reduces the risk of specimen
contamination by microorganisms (e.g.,
blood cultures) and additive carry-over,
which affects some chemistry tests.
• Additive carry-over can occur when
blood in an additive tube touches the
needle during venipuncture or during
transfer from a syringe. Additive in the
blood that is on or within the needle can
then be transferred to the next tube
drawn or filled.
• The most common tubes in the order of
draw can be remembered by recalling
the phrase "stop, light red, stay, put,
green light, go"
• The first letter of each word in the
phrase stands for a tube in the order of
draw: S (sterile), L (light blue), R (red), S
(serum separator tube or SST), P
(plasma separator tube or PST), G
(green), L (lavender), and G (gray).
Syringe System
Sometimes used for patients with small or
difficult veins. This system consists of a sterile
syringe called a hypodermic needle and a
sterile plastic syringe.
Syringe Needle- those appropriate for
phlebotomyare generally gauged 21 to 23, in 1
or 1.5in. lengths.
Syringe- most common volumes for
phlebotomy are 2, 5, and 10 mL. Syringe
volume is selected according to the size and
condition of the patient’s vein anf the amount
of blood to be collected.

This system consists of a sterile syrunge needle

called a hypodermic needle and asterile plastic
syringe with a Luer-lock tip.

Syringes have two parts:

1. Barrel- cylinder with graduated
markings in either mL or cc
2. Plunger- rod-like device that fits tightly
into the barrel
When drawing venous blood by syringe, the
plunger is pulled back, creating a vacuum that
fills the barrel with blood.

Winged Infusion Set

A winged infusion blood collection set, or
butterfly, is an indispensable tool for collecting
blood from small or difficult veins such as hand
veins and veins of elderly and pediatric
patients as it allows much more flexibility and
precision than a needle and syringe.
It consists of a 1/2- to 3/4-inch stainless steel
needle permanently connected to a 5- to 12-in.
length of tubing with either a Luer attachment
for syringe use or a multisample Luer adapter
foruse with the evacuated tube system.
• 23-gauge needle is most commonly
used for phlebotomy.
• 25-gauge needle is used by specially
trained personnel to collect blood from
scalp or other tiny veins of premature
infants and other neonates.