PRINCIPLES OF MEDICAL LABORATORY SCIENCE 2

College of Allied Health Sciences

Bachelor of Science in Medical Laboratory Science
Second Semester, A.Y. 2022-2023

[TRANS] PMLS UNIT 4: BLOOD COLLECTION EQUIPMENT AND SUPPLIES, ADDITIVES AND ORDER OF DRAW
o Povidone-iodine(0.1%-1% available
OUTLINE iodine)
o Tincture of Iodine
I Blood Collection Equipment  Disinfectants
II Tube Additives - chemical substances regulated by the
III Order of Draw Environmental Protection Agency (EPA)
- are used to remove or kill microorganisms on
surfaces and instruments
- are corrosive and are not safe to use on human
BLOOD COLLECTION EQUIPMENT skin
- EPA-registered sodium hypochlorite products are
BLOOD-DRAWING STATION preferred
 Blood Drawing Station o solutions made from generic 5.25%
o A dedicated area of a clinical laboratory equipped sodium hypochlorite (household bleach)
for performing phlebotomy procedures on patients, may be used
primarily outpatients sent by their physicians for o 1:100 → nonporous surfaces after
laboratory testing. cleaning up blood or other body fluid
o Phlebotomy Chair spills
o 1:10 → is applied prior to cleanup when
spills involve large amounts of blood or
other body fluids
o At least 10 minutes of contact time
 Hand Sanitizers
- CDC/HICPAC recommends the use of alcohol-
based hand sanitizers for routine
decontamination of hands (when hands are not
visibly soiled).
- detergent-containing wipes followed by the use
of an alcohol-based hand cleanser → if hands are
heavily contaminated with organic material
and hand-washing facilities are not available
 Gauze Pads/Cotton Balls
- Clean 2-by 2-inch gauze pads folded in fourths
are used to hold pressure over the site following
 Equipment Carriers blood collection procedures
o Handheld Carrier (Phleb Kit) - Use of cotton balls to hold pressure is not
o Phlebotomy Cart recommended
BLOOD COLLECTION EQUIPMENT  Bandages
 Gloves - Are used to cover a blood collection site after the
- The CDC/HICPAC standard precautions and bleeding has stopped
OSHA require the wearing of gloves when - CAUTION: Adhesive bandages should not be
performing phlebotomy. used on babies younger than 2 years of age
- A new pair must be used for each patient and be because of the danger of aspiration and
removed when the procedure is completed. suffocation.
- Nonsterile, disposable latex, nitrile, neoprene,  Needle and Sharps Disposal Containers
polyethylene, and vinyl examination gloves - sharp objects must be disposed of immediately in
 Antiseptics special containers referred to as “sharps”
- Substances used to prevent or inhibit the containers
growth and development of microorganisms but  Biohazard Bags
do not necessarily kill them - leakproof plastic bags that are commonly used
- Safe to use on human skin and are used to clean to transport blood and other specimens from the
the venipuncture site collection site to the laboratory
- most commonly used: 70% isopropyl alcohol  Slides
(isopropanol) - Precleaned 25- by 75-mm (1- by 3-in.) glass
o Benzalkonium chloride (Zephiran microscope slides
chloride, 1:750) o are used to make blood films for
o 70% isopropyl alcohol followed by an hematology determinations
iodophor o either plain or with a frosted area at one
o Chlorhexidine gluconate end
o 70% Ethyl Alcohol  Pen
o Chlorhexidine gluconate - A phlebotomist should always carry a pen with
o Hydrogen Peroxide indelible (permanent) nonsmear ink to label tubes
and record other patient information.

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 TRANS: PMLS 2 Unit 4

 Watch 20 Multisample Sometimes used when

- A watch, preferably with a sweep second hand or Hypodermic large-volume tubes are
timer, is needed to accurately determine collected or large-
specimen collection times and time certain volume syringes are
tests. used on patients with
VENIPUNCTURE EQUIPMENT normal-size veins
 Evacuated Tube System (ETS) – most preferred 21 Multisample Considered the
 Syringe System – used on small, fragile and damaged Hypodermic standard
veins venipuncture needle
 Butterfly System – can be used with the ETS and syringe. for routine
It is used to draw blood from infants and children’s hand venipuncture on
veins and other difficult-draw situations patients with normal-
VEIN LOCATING DEVICES size veins
 Vein-Locating Devices – transillumination devices that 22 Multisample Used on older children
make it easier to locate veins that are difficult to see or feel Hypodermic and adult patients with
 Tourniquet – A device that is applied or tied around a small veins or for
patient’s arm prior to venipuncture to restrict blood flow syringe draws on
difficult veins
o Strap Tourniquet – most common type; is a flat
strip of stretchable material, such as latex, nitrile, or 23 Butterfly Used on the veins of
vinyl that is fairly inexpensive and disposable infant and children and
o 1 minute Application on difficult or hand
veins of adults
NEEDLES
 Multisampel Needles – needle color codes vary among
 Needles – are sterile, disposable, and designed for a
manufacturers; however, several large manufacturers use:
single use only
o Yellow for 20-gauge
o Multisample needles – allows collection of multiple
o Green for 21-gauge
tubes during venipuncture
o Black for 22-gauge
o Hypodermic needles – used for patients with veins
from which it is difficult to collect blood and for  Tube Holder – plastic cylinder with a small opening for a
blood gas analysis. needle at one end and a large opening for tubes at the other
o Winged infusion (butterfly) needles – an end
indispensable tool for collecting blood from small or o OSHA regulations require that the tube holder with
difficult veins such as hand veins and veins of needle attached be disposed of as a unit after use
elderly and pediatric patients as it allows much more and never be removed from the needle and reused.
flexibility and precision than a needle and syringe. o A needle or tube holder that has a safety device is
an example of a SESIP, which is the OSHA acronym
PARTS OF A NEEDLE
for a sharp with engineered sharps injury
 Bevel – the end that pierces the vein, it is “beveled,” or
protection.
cut on a slant
EVACUATED TUBES
o The bevel is the slanted opening at the end of the
needle.  Evacuated Tubes
o Bevel of the needle must face upward when the - Designed to fill with a predetermined volume of
needle is inserted into the vein. blood by vacuum
- Rubber stoppers (color coded according to
 Shaft – the long cylindrical portion
additive content)
 Hub – the end that attaches to the blood collection
o Premature Loss of Vacuum → PARTIAL DRAW
device
OR SHORT DRAW:
 Bore/Lumen – internal space of the needle
 Improper storage
NEEDLE GAUGE  Opening the tube
 Needle Gauge – indicated by a number that is related to  Dropping the tube
the diameter of the lumen  Advancing the tube too far onto the needle
o the higher the gauge number, the smaller the before venipuncture
actual diameter of the needle  If the needle bevel becomes partially out of
o 21 G – standard for venipuncture the skin during venipuncture
o 23 G – for children; small and difficult veins  Stoppers
o 25 G – used by specially trained personnel to collect - Also known as tops or closures
blood from the scalp or other tiny veins of - Are made up of rubber
premature infants and neonates - Some tubes have a rubber stopper covered by a
Gauge Needle Type Typical Use plastic shield
15-17 Special needle Collection of donor Stopper Color Additive Department(s)
attached to collection units, autologous blood Light Blue Sodium Coagulation
bag donation, and Citrate
therapeutic phlebotomy Red None Chemistry, Blood Bank,
18 Hypodermic Used primarily as a Serology/Immunology
transfer needle rather Red Clot Activator Chemistry
than for blood Red/Light None NA (Discard Tube only)
collection; safety Gray
tissues have diminished
Clear
use

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 TRANS: PMLS 2 Unit 4

Red/Black Clot Activator Chemistry  CLSI recommends spray-dried EDTA for most hematology
(Tiger) and Gel tests
Gold Separator
Red/Gold KEY POINT: Excess EDTA, which results when tubes are
Green/Gray Lithium Chemistry underfilled, can cause RBCs to shrink and thus change
Light Green Heparin and CBC results!
Gel Separator CITRATES
Green Lithium Chemistry  Prevent coagulation by binding or chelating calcium
Heparin  Most common citrate is sodium citrate, which is used for
Sodium coagulation tests
Heparin  Sodium citrate tubes have light-blue stoppers (B:A = 9:1)
Ammonium  Sodium citrate – also used in special Erythrocyte
Heparin Sedimentation Rate (ESR) tubes with black stoppers
Lavender EDTA Hematology (B:A = 4:1)
(Purple)  3-4 gentle inversions are required for adequate mixing
Pink EDTA Blood Bank HEPARIN
Gray Sodium Chemistry  Prevents clotting by inhibiting thrombin formation
fluoride and  Heparinized plasma is often used for some chemistry
potassium tests, especially stat tests (e.g., electrolytes)
oxalate
 Heparin is the additive in:
Sodium
o Green-top tubes
fluoride and
o Green-top and light green–top gel tubes
EDTA
o Mottled-green and gray-top tubes
Sodium
o Royal blue–top tubes with green color coding on
fluoride
the label
Orange Thrombin Chemistry o Green-top and light green–top microcollection
Gray/Yellow tubes
Royal Blue None Chemistry o Red-banded and green-banded microhematocrit
EDTA tubes
Sodium  Three heparin formulations:
heparin o Ammonium
Tan EDTA o Lithium
Yellow Sodium Microbiology o Sodium heparin
polyanethol  5-10 inversions
sulfonate
OXALATE
(SPS)
 Prevents coagulation by precipitating calcium
Yellow Acid citrate Blood
dextrose Bank/Immunohematology  Potassium oxalate is the most widely used
(ACD)  It is commonly added to tubes containing glucose
preservatives to provide plasma for glucose testing
 Potassium oxalate is the additive in:
BLOOD COLLECTION EQUIPMENT
o Gray-top evacuated tubes and
o Microcollection containers with gray tops
ANTICOAGULANTS
 8-10 inversions are required for proper mixing
 Anticoagulants – substances that prevent blood from
SPECIAL USE ANTICOAGULANTS
clotting (coagulating) by either of two methods
o by chelating (binding) or precipitating calcium  Acid-Citrate Dextrose(ACD)
o by inhibiting the formation of thrombin o ACD solution is available in two formulations
(solution A and solution B) for
 Specimen – whole blood or Plasma
immunohematology tests such as DNA testing and
human leukocyte antigen (HLA) phenotyping, which
EDTA (ETHYLENEDIAMINETETRAACETIC ACID) is used in paternity evaluation and to determine
 Commonly available as a powdered K2EDTA or liquid transplant compatibility.
K3EDTA  The acid citrate prevents coagulation by
 Prevents coagulation by binding or chelating calcium binding calcium.
 EDTA is the additive in:  Dextrose acts as a RBC nutrient and
o Lavender (purple)-top tubes preservative by maintaining RBC viability.
o Microcollection containers with lavender tops o ACD tubes have yellow tops and require 8
o Pink plastic-top tubes with a special blood bank inversions immediately after collection to prevent
patient ID label clotting
o Pearl-top tubes with thixotropic gel separator  Citrate Phosphate Dextrose (CPD)
o Royal blue–top tubes with lavender color-coding o used in collecting units of blood for transfusion
on the label  Citrate prevents clotting by chelating
 Provide whole-blood specimens for hematology tests calcium
 8-10 inversions are normally required for proper mixing  Phosphate stabilizes pH
 Caution: If microclots are detected in a hematology  Dextrose provides cells with energy and
specimen, it cannot be used for testing and must be helps keep them alive
recollected.  Sodium Polyanethol Sulfonate (SPS)

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 TRANS: PMLS 2 Unit 4

o Prevents coagulation by binding calcium 2. Blue-top coagulation tube

o Used for blood culture collection 3. Serum tube with or without clot activator, with or
o It also reduces the action of a protein called without gel
complement, which destroys bacteria 4. Heparin tube with or without gel plasma
o Yellow stoppers separator
o Requires 8 inversions to prevent clotting 5. EDTA tube
6. Glycolytic inhibitor tube
ANTIGLYCOLYTIC AGENTS
 Antiglycolytic Agents  STOP, LIGHT RED, STAY PUT, GREEN LIGHT, GO
o A substance that prevents glycolysis
o Sodium fluoride - most common antiglycolytic
agent
o Glucose is metabolized at RT at a rate of 7
mg/dL/hour and at 4˚C, glucose decreases by
approximately 2 mg/dL/hour.
o Preserves glucose for up to 3 days and also inhibits
the growth of bacteria
o Gray stoppers
o Requires 5-10 inversions
CLOT ACTIVATORS
 Clot Activators
o are used to collect serum specimens
o include substances such as glass (silica) particles
and inert clays like Celite, and clotting factors
such as thrombin
o Silica particles – are the clot activators in
serumseparator tubes (SSTs) and plastic red-top
tubes; cause the blood to clot within 15 to 30
minutes
o Thrombin tubes - clot within 5 minutes
o Require 5 gentle inversions
THIXOTROPIC GEL SEPARATOR
 Thixotropic Gel Separator
o An inert (nonreacting) synthetic substance
initially contained in or near the bottom of certain
blood collection tubes
o Serum gel-barrier tubes are called serum-separator
tubes (SSTs)
o Heparinized plasma gel-barrier tubes are called
plasma-separator tubes (PSTs).
TRACE ELEMENT-FREE TUBES
 Trace Element-Free Tubes
o Have royal-blue stoppers
o Are made of materials that are as free of trace
element contamination as possible
o They are used for trace element tests, toxicology
studies, and nutrient determinations
o Royal blue–top tubes contain EDTA, heparin, or no
additive to meet various test requirements

ORDER OF DRAW
 Order of Draw
o Refers to the order in which tubes are collected
during a multiple-tube draw or are filled from a
syringe
o A special sequence of tube collection that reduces
the risk of specimen contamination by
microorganisms
 Additive Carry Over
o Affects chemistry test
o Occurs when blood in an additive tube touches
the needle during venipuncture or during transfer
from a syringe

CLSI RECOMMENDED ORDER OF DRAW

 CLSI recommends the following order of draw:
1. Sterile tube (blood culture)

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