Lesson 1: Laboratory Safety and Hazards 4.

Radioactive
○ Source: Equipment and radioisotopes
Laboratory Safety ○ Possible Injury: Radiation exposure
● Safety begins with the recognition of hazards and is
achieved through the following: 5. Electrical
○ Application of common sense ○ Source: Undergrounded or wet equipment;
○ Listen to the instructions frayed cords
○ A safety-focused attitude ○ Possible Injury: Burns or shock
○ Good personal behavior
○ Good housekeeping in all laboratory work and 6. Fire / Explosive
storage areas ○ Source: Bunsen burners, organics chemicals
○ Continual practice of good laboratory ○ Possibly Injury: Burns, dismemberment
technique
Two Primary Causes of Accidents: 7. Physical
● Unsafe activities ○ Source: Wet floors, heavy boxes, patients
○ Storage in the lab should be in appropriate ○ Possibly Injury: Falls, sprain or strains
places
○ Chemical & solutions Types of Safety Hazards (Elaborated)
■ Labels must be classify 1. Biological Hazard
● Information must be complete ○ Refers to biological substances that pose a
● Labeling tags must not easily threat to the health of living organisms,
fade or easily to tear primarily that of humans.
○ These microorganisms are frequently present
● Unsafe environmental conditions in the specimens received in the clinical
○ Temperature laboratory.
○ Air quality
○ Lighting ○ Infection control
■ healthcare facilities developed
Safety Equipment procedures to control and monitor
● Safety showers & eye wash station (required) infections occurring within the facilities.
● Fire extinguisher
● Fume Hood ○ CDC (Centers for Disease Control and
○ Prepare chemical in mixing solution Prevention)
○ Releases fume coming from chemicals ■ Universal Precautions(1985)
● Biosafety cabinets ● Blood and body fluid
○ Handle microorganism precautions should be
● Personal Protective Equipment consistently used for all patients
○ Laboratory gown
○ Laboratory coat Six Component of the Chain of Infection
○ Mask 1. Infectious Agents
○ Shoe covers ○ Pathogenic microbe
○ Disposable laboratory gown ■ Virus
○ Eye goggles ■ Bacteria
○ Gloves ■ Fungi
■ Parasites
Types of Safety Hazard ■ Viruses
1. Biological 2. Reservoir
○ Source: Infectious agents ○ Location of potentially harmful microorganism
○ Possible Injury: (Infections) bacterial, fungal, ■ Contaminated clinical specimen or an
viral, or parasitic infected patient
○ Place where the infectious agent can live and
2. Sharps possible multiply
○ Source: Needles, lancets, broken glass ■ Humans (excellent reservoirs)
○ Possible Injury: Cuts, punctures or ■ Animals (excellent reservoirs)
blood-born pathogen exposure ■ Food
■ Water
3. Chemical ■ Soil
○ Source: Preservative and reagents ■ Equipment
○ Possible Injury: Exposure to toxic,
carcinogenic, or caustic agents
 ○ Fomites Additional Note:
■ Reservoirs coming from equipment ● Laboratory and health workers are highly exposed to
■ These are equipment and other soiled infections (pathogens)
inanimate objects will serve as ● Pathogens in hospitals are more violent
reservoirs, particularly if they contain: ○ Infections contracted in hospital
● Blood ○ Hospital acquired infection
● Urine ○ Nosocomial infection are due to poor hygiene
● Other body fluids ● 2 type of Infection
○ Communicable Infections
3. Portal of exit ○ Nosocomial Infections and
○ Way of exit of reservoir to continue the chain healthcare-associated infections (HAIs)
of infection.
○ This can be through the: 3 Primary Element of Chain of Infection
■ Mucous membranes of the 1. Reservoir
● Nose 2. Mode of transmission
● Mouth 3. Susceptible Host
● Eyes
■ In blood or other body fluids. How to break the link of Chain of Infection

4. Means of Transmission
○ Direct contact:
■ the unprotected host touches the
patient, specimen, or a contaminated
object
○ Airborne:
■ inhalation of dried aerosol particles
circulating on air currents or attached
to dust particles
○ Droplet:
■ the host inhales material from the
reservoir
○ Vehicle:
■ ingestion of a contaminated substance
○ Vector:
■ from an animal or insect bite

5. Portal of Entry
○ Way of entry of reservoir to continue the chain
of infection.
○ Same as the portal of exit Chain of Infection
○ This can be through the: ● Major importance in the laboratory.
■ Nose ○ Proper hand hygiene
■ Mouth ○ Correct disposal of contaminated materials
■ Eyes ○ Wearing personal protective equipment (PPE)
■ breaks in the skin OSHA
■ Open wounds. ● Stands for “Occupational Safety Health
Administration”
6. Susceptible Host ● Federal agency in US
○ Can be another patient during invasive ○ Committed to safeguarding worker health and
procedures, visitors, and health care safety
personnel when exposed to infectious
specimens or needlestick injuries OSHA Blood-Borne Pathogens standard requires written
○ Someone who is prone to infection “Exposure Control Plan”
■ Elderly Categories of Exposure:
■ Newborn babies a. Category I – daily exposure to blood and body fluids
■ Patients who are immune-suppressed b. Category II – regular exposure to blood and body
or unvaccinated fluids
■ Those suffering from acute or chronic c. Category III – no exposure to blood and body fluids
illness
Take note:
● Employers must offer HBV vaccine (known as Hepa B
vaccine) to all personnel
● Required for category I and II
 3. Reactive Chemicals
Nice to Know ○ Spontaneously explode or ignite or the evolve
● Specimens should be “capped” during heat or flammable or explosive gasses.
centrifugation
● Any blood, body fluid, or other potentially 4. Chemical Labeling
infectious material spill must be cleaned up ○ Hazardous chemicals should be labeled with a
using: description of their particular hazard, such as:
● Spill cleanup kit, Common aqueous detergent, ■ Poisonous
10% bleach using appropriate contact time ■ Corrosive
● Recommended contact time is 10 minutes ■ Flammable
■ Explosive
2. Chemical Hazard ■ Teratogenic or carcinogenic
○ Can cause skin burns and irritation
MATERIAL SAFETY DATA SHEETS (MSDS)
○ Chemical spills and Exposure: 1. Physical and chemical characteristics
■ When skin contact occurs, 2. Fire and explosion potential
● The best first aid is to flush the 3. Reactivity potential
area with large amounts of 4. Health hazards and emergency first aid procedures
water for at least 15 minutes 5. Methods for safe handling and disposal
and then seek medical 6. Primary routes of entry
attention. 7. Exposure limits and carcinogenic potential

○ Chemical Handling: 3. Fire Hazards

■ Chemicals should never be mixed ○ The Joint Commission on Accreditation of
together unless specific instructions Healthcare Organizations (JCAHO)
are followed ■ requires that all health-care institutions
■ they must be added in the order post evacuation routes and detailed
specified. plans to follow in the event of a fire.
■ This is particularly important when ○ Laboratory personnel should be familiar with
combining acid and water. these procedures. When a fire is discovered,
● In adding acid to water it should all employees are expected to take the actions
be gradual in the acronym :
■ Race
○ Chemical Hygiene Plan: ■ Alarm
■ OSHA also requires all facilities that ■ Contain
use hazardous chemicals to have a ■ Extinguish/ Evacuate
written chemical hygiene plan (CHP) ○ The National Fire Protection Association
available to employees. (NFPA)
■ The purpose of the plan is to detail the ■ has developed the Standard System
following: for providing codes and standard
● Appropriate work practices information about the
● Standard operation procedures chemicals/solutions.
● PPE ■ NFPA 704 - Hazardous Material
● Engineering controls Symbol
○ such as fume hoods and
flammable safety
cabinets
● Employee training equipment
● Medical consultation
guidelines.

Storage and Handling of Chemicals

1. Flammable/Combustible Chemicals
○ Classified according to flash point
○ Flash point
■ The temperature at which sufficient
vapor is given off to form an ignitable
mixture with air.

2. Corrosive Chemicals
○ Injurious to the skin or eyes by direct contact
or to the tissue of the respiratory and
gastrointestinal tract if inhaled or ingested
 7. Physical Hazards
○ Physical hazards are not unique to the
laboratory, and routine precautions observed
outside the workplace apply.
○ General precautions to consider are:
■ to avoid running in rooms and hallways
■ watch for wet floors
■ bend the knees when lifting heavy
objects
■ keep long hair pulled back
■ avoid dangling jewelry
■ and maintain a clean
■ organized work area.
■ Closed-toed shoes that provide
maximum support are essential for
safety and comfort.

8. Mechanical Hazards
○ Centrifuges
■ must be balanced to distribute the load
equally.
■ Never open the lid until the rotor has
come to a complete stop

Disposal of Hazardous Materials

1. Biological/Biohazardous Waste
○ - All biological waste (EXCEPT URINE) should
4. Radioactive Hazards be placed in appropriate containers labeled
○ Equipment and radioisotopes with biohazard symbols.
○ Radiation Safety ■ URINE: may be discarded by pouring it
○ All areas where radioactive materials are used into the lab sink.
or stored must be posted with caution signs, ● The sink should be flashed also
and traffic in these areas should be restricted with water after the urine has
to essential personnel only. been discarded. -
○ Exposure to radiation during pregnancy ○ Decontaminate the sink by 1:5 or 1:10 dilution
presents a danger to the fetus, and personnel of sodium hypochlorite (bleach solution).
who are or who think they may be pregnant ○ Disinfection of the sink should be performed
should avoid areas with this symbol. daily.
○ The symbol must be displayed on the doors of ■ Empty urine containers can be
all areas where radioactive material is present. discarded as non biologically
hazardous waste.
5. Electrical Hazards ○ Incineration, inactivation, burial, chemical
○ Equipment should not be operated with wet disinfection, encapsulation in a solid matrix
hands.
○ Designated hospital personnel monitor
electrical equipment closely
■ However, laboratory personnel should
continually observe for any dangerous
conditions, such as frayed cords and
overloaded circuits
○ Equipment that has become wet should be
unplugged and allowed to dry completely
before reusing.
■ Equipment also should be unplugged
before cleaning.
○ All electrical equipment must be grounded with
three-pronged plugs.

6. Sharps Hazard
○ Pertains to needles, lancets, broken
glassware/glass slides
○ All sharp objects must be disposed in
puncture-resistant, leak-proof container with
the biohazard symbol
 2. Chemical Waste Lesson 2: Introduction to Phlebotomy
○ Flush water
■ soluble substances down the drain with History of Phlebotomy
large quantities of water ● Process of collecting blood through the vein​
○ Strong acids and bases should be neutralized ● For analysis and diagnostic
before disposal ○ requested by the Physician)
○ Foul smelling chemicals should never be ● Greek words:
disposed down the drain ○ Phlebos (vein)
○ Flammable solvents ○ Temnein (cut) ​
■ collected in approved containers ● Venesection​
○ Flammable material ○
■ specially designed incinerators ● 1400 BC​
○ Solid chemicals ○ Ancient Egyptians (bloodletting)
■ Landfill
● Hippocrates (Greek Physician) –
Waste Disposal Technique ○ four humors:
● Incineration ■ Earth - Blood and brain
● Recycling ■ Air - Phlegm and lungs
● Landfill burial ■ Water - Yellow bile and gallbladder
● Flushing down the drain ■ Fire - Black bile​and spleen
● Bloodletting (removal of the excess humor) ​
● Middle Ages
○ Barber-surgeons performed bloodletting
(treatment for some illnesses) ​
● 17th and 18th century
○ phlebotomy was treated as major therapy​
● Cupping and leeching (lancets and fleams) ​
● Cupping
○ Ease pain, inflammation and other
health-related concerns
○ application of heated suction cups on the
patient’s skin and incision ​
● Leeching (hirudotherapy) –
○ Method uses leeches for bloodletting and is
currently used for microsurgical replantation
○ involves placing a drop of milk or blood on
patient’s skin and introducing Hirudo
medicinalis to the site, allowing it to engorge or
fall off

● Main goals: ​
○ Diagnosis and treatment using blood samples​
○ Transfusion (remove blood from donor)​
○ Removal of blood for polycythemia
(polycythemiavera) or therapeutic purposes
○ Polycythemiavera - condition where the bone
marrow goes into “overdrive” and makes too
many red blood cells, along with too many
white blood cells and platelets.

● Main methods: ​
○ Venipuncture
■ Large vein
■ needle inserted in a vein​
○ Capillary
■ Small vein
■ puncturing the skin
■ Skin / dermal
Roles of the Phlebotomists Professional and Personal Characteristics of a
● Collect blood samples (properly labeled) ​ Phlebotomist
● Delivering or transporting collected samples 1. Dependable, cooperative, committed
(prescribed time limits)​ 2. Compassionate, courteous, respectful
● Process collected samples (centrifuging and 3. Integrity, honesty, competence
aliquoting [smaller])​ 4. Organized, responsible, flexible
● Assist in collecting other specimens (urine; non-blood 5. Appearance (Clothing, Hygiene)
samples)​ 6. Communication
● Drawing of blood units from donors for transfusion to ○ Verbal Skills
a patient requiring whole blood or blood components ○ Nonverbal Skills
○ Body language
7. Respecting Cultural Diversity
8. Telephone skills

The Healthcare Setting

● Healthcare facilities category:
○ Inpatient (non-ambulatory)
■ Confinement and admission of the
patient
■ tertiary care practitioners
○ Outpatient
Duties of a Phlebotomist ■ Check-ups
● Traditional Duties ■ secondary care specialists
1. Correct identification and preparation of the
patient before sample collection. All patients can be serviced by primary, secondary, and
2. Collection of the appropriate amount of blood tertiary levels of healthcare (Williams-tungpalan, 1981)
by venipuncture or dermal puncture for the ● Primary level
specified tests. ○ health units in the rural areas and sub-units
3. Selection of the appropriate sample containers (DOH)
for the specified tests. ● Secondary level
4. Correct labeling of all samples with the ○ non-departmentalized hospitals that attend to
required information. patients during the symptomatic stages of an
5. Appropriate transportation of samples back to ailment.
the laboratory in a timely manner. ○ district , community, regionals
6. Effective interaction with patients and hospital ● Tertiary level
personnel. ○ medical centers and large hospitals; can
7. Processing of samples for delivery to the address serious diseases
appropriate laboratory departments. ○ High–end, specialized
8. Performance of computer operations and ● Ambulatory care
record-keeping pertaining to phlebotomy. ○ medical care given to outpatients or patients
9. Observation of all safety regulations, quality requiring care or follow-up check-ups after
control checks, and preventive maintenance discharge from the hospital (freestanding
procedures. medical care setting; hospital-owned clinics;
10. Attendance at continuing education programs outpatient departments and urgent care
facilities)
● Additional Duties ● Homebound services
1. Training other health-care personnel to ○ procedures, tests, and services (patient’s
perform phlebotomy home; long-term facility)
2. Monitoring the quality of samples collected on ○ Monitoring vital signs
the units ● Public Health Services
3. Evaluation of protocols associated with sample ○ Services are offered with little or no charge at
collection all
4. Performing and monitoring point-of-care
testing (POCT) The Clinical Laboratory
5. Performing electrocardiograms ● Hospital
6. Performing measurement of patient’s vital ○ institution that has permanent inpatient beds
signs with 24-hour nursing service along with
7. Collection of arterial blood samples therapeutic and diagnostic service
8. Collection of samples from central venous ○ it houses the clinical laboratory services where
access devices(CVADs) tests requested by Physicians are performed
● 2 major divisions:
○ anatomical and surgical pathology
○ clinical pathology
Clinical Pathology / Clinical Analysis Chemistry - Glucose
● Hematology ● Amounts of certain chemicals in a blood sample and
● Coagulation evaluate level indicates diabetes mellitus
● Chemistry ● Fasting Blood Sugar (FBS)
● Serology/Immunology ● Glucose Tolerance Test (GTT)
● Urinalysis ● 2-hour Post Prandial Blood Sugar (2-h-PPBS)
● Microbiology ● Random Blood Sugar (RBS)
● Blood Bank/ Immunohematology ● Glycosylated Hemoglobin (HbA1c)

Anatomical and Surgical Pathology Chemistry – Lipid Profile/Panel

● Histology ● Total Cholesterol (Ch)
○ Microscopic structure of tissues ● Triglycerides (TAG)
● Cytology ● High Density Lipoprotein (HDL)
○ Structure of cells ● Low Density Lipoprotein (LDL)
● Cytogenetics ● Very Low-Density Lipoprotein (VLDL)
○ Chromosomal deficiencies and genetic
disease Chemistry – Metabolic Panel/Profile
Hematology ● General test for metabolic screen
● Analyzes blood for diseases ○ Blood Urea Nitrogen
○ Erythrocytes (RBC) ○ Creatinine (kidney)
○ Leukocytes (WBC) ○ Blood Uric Acid
○ Platelets ○ Electrolytes (NA+, Ca+, K+, Cl+, Mg+)
○ plasma
● Whole blood samples Chemistry – Other tests
○ EDTA – violet (general) ● Creatine Kinase (CK)
○ Citrate – blue (coagulation tests) ● Creatine Kinase-MB (CKMB)
● Pleural (liquid pleura, in thoracic) ● Lactate dehydrogenase (LDH)
● Pericardial (excess, in heart) ● Acid Phosphatase (ACP)
● Peritoneal (abdominal) ● Gamma glutamyl transferase (GGT)
● Synovial (joint fluid) ● Troponin T and I (Trop T) (Trop I)
● Spinal (cellular) ● Hormones

Hematology - Complete Blood Count (CBC) Serology/Immunology

● Evaluate anemia, infections, malignancy and tumors ● Patients’ immune response
○ Hematocrit (Hct) ● Serum separator tube (red top)
○ Hemoglobin (Hb) ○ Anti-Nuclear Antibodies (ANA)
○ Red Blood Cell (RBC) Count ■ Autoimmune disease
○ White Blood Cell (WBC) Count ○ HIV testing – Anti HIV
○ Platelet Count ○ Syphilis testing – RPR (Rapid Plasma Reagin)
○ Differential WBC Count ○ Hepatitis testing - HBsAg, HCV
○ Indices ○ Typhidot (typhoid fever)
○ Mean Corpuscular Hemoglobin (MCH) ○ Anti-Streptolysin O (ASO) (streptococcal
○ Mean Corpuscular Volume (MCV) infections)
○ Mean Corpuscular Hemoglobin Concentration ○ Complement levels
(MCHC) ○ Rheumatoid Factor (Rheumatoid arthritis)
○ RBC Distribution Width (RDW)
○ Hematology – Peripheral Blood Smear (PBS) Clinical Microscopy
○ Hematology – Erythrocyte Sedimentation Rate ● Urinalysis (U/A)
(ESR) ○ Kidney diseases and metabolic disorders
○ Hematology – Reticulocyte Count ○ Reagent strips /dipsticks
○ Hematology – Erythrocyte Osmotic Fragility ■ Chemicals embedded in color pads
Test (EOFT) (color changes)
○ Hematology – Other body fluids microscopy ● Fecalysis (F/A)
○ Hematology – Bone Marrow Smears ● Fecal Occult Blood Test (FOBT)
○ Physical - color, clarity
Coagulation ○ Chemical - glucose, pH, protein, nitrate,
● Ability of blood to form and dissolve clots bilirubin, urobilinogen
○ Activated Partial Thromboplastin (APTT) ○ Microscopic - RBC,WBC, cast, cells
○ D – dimer
○ Fibrin Split Products (FSP)
○ Fibrinogen Test
○ Prothrombin Time (PT)
Microbiology Lesson 3: Venipuncture Equipment
● Isolate and identifies pathogens
● Bacteriology, urology, mycology (fungi), parasitology Requisition Form
○ Gram’s Stain (GS) ● CLSI Mandate:
■ Bacteria ○ Clinical Laboratory Standards Institute
● Gram (+), (-) ○ Verify patient Information from at least 2
○ Acid – Fast Bacilli Staining (AFB) sources
■ Grows microorganism and determines
antibiotic Organization of Equipment
○ Culture and Sensitivity (C&S)
■ Grows microorganism and determines
antibiotic
○ Ova and Parasite (O&P)
● Bacterial cultures
● Modern molecular diagnostics

Blood Bank/ Immunohematology

● For blood transfusions
● Red top tubes and EDTA - pink top tubes
○ Blood Typing
○ Antibody Screening (unexpected)
○ Antibody Identification (identifies unexpected)
○ Compatibility testing – Crossmatching
○ Antiglobulin Test or Coomb’s Test – Direct and
Indirect Coomb’s
○ Donor recruitment, donor selection (eligibility)
○ Donor Screening

STAT LABS
● This facility is usually located near the emergency
Evacuated Tube System
room of some tertiary care facilities so that
● Consists of a double-pointed needle to puncture the
procedures and tests can be done immediately when
stopper of the collection tube, a holder to hold the
needed
needle and blood collection tube, and color-coded
evacuated tubes.
Reference Laboratory
● Large and independent laboratory that provides
Needles
specialized and confirmatory laboratory tests for
● Venipuncture needles include multisample needles
blood, urine, and tissues, and offers as well faster
(ETS), hypodermic needles, and winged blood
turnaround or processing time
collection needles (butterfly).
● Sterile, disposable, and are used only once
● Needle size varies by both length and gauge
(diameter)
○ For routine venipuncture:
■ 1-inch and 1.5-inch lengths are used
○ Needles gauge refers to the diameter of the
needle bore
○ Needles vary from large (16-gauge) needles
used to collect units of blood for transfusion to
much smaller (23-gauge) needle used for very
small veins
● The smaller the gauge number the bigger the
diameter of the needle
Multisample needles Most Common Anticoagulant work by binding calcium
● Have the stopper puncturing needle covered by a ● Ethylenediamine tetraacetic acid (EDTA)
rubber sheath that is pushed back when a tube is ○ Chilates calcium
attached and returns to full needle coverage when the ● Citrates
tube is removed. This prevents leakage of blood when ● oxalates
tubes are being changed
● All needles consist of a bevel (angled point), shaft, Heparin
lumen, and hub ● prevents clotting by inhibiting the formation of
thrombin necessary to convert fibrinogen to fibrin in
the coagulation process

Evacuated Tubes: Serum (clotted blood) Tubes

● Red top
○ Glass tubes
■ No clot activator
■ No anticoagulant
■ Clear
○ Plastic tubes
■ With clot activator and gel separator
■ No anticoagulant
Needle Disposal System ■ Hazy
● Needles with safety devices activated must always be ○ Used for
placed in ■ ‘Routine’ Clinical Chemistry test
○ Rigid ● electrolytes
○ Puncture-resistant ● Glucose
○ Leak-proof disposable “sharps” containers ● Renal
labelle biohazard that are easily sealed and ● Liver
locked when full. ● Thyroid function lipids
● Cardiac markers
Needle/ Tube Holder ■ Immunochemistry
● Made of rigid plastic and may be designed to act as a ● Hormones
safety shield for the used needle. ● Vitamins
■ Serology

Evacuated Tubes: Plasma Tubes

● EDTA
○ Lavender or pink top
○ Dry di-potassium (K2) amd liquid tri-potassium
(K3)
○ Used for
■ Hematology test
■ HBA1c
■ Hemoglobin electrophoresis
■ Reticulocyte counting
■ Pink - for Blood bank (crossmatching
Collection Tubes and ABO typing)
● Tubes used for blood collection
● Contain a premeasured amount of vacuum ● Sodium Citrate
● Are available in glass and plastic ○ Blue top
● May also contain anticoagulants and additives ○ 3 concentrations:
● Labeled w/ type of anticoagulant or additive, the draw ■ 0.105 mol/L or 3.13% (glass)
volume and the expiration date ■ 0.109 mol/L or 3.2 % (plastic)
■ 0.129 mol/L or 3.8% (plastic)
Anticoagulant tubes ○ 1:9 citrate to blood ratio
● Test requiring whole blood or plasma are collected in ○ Used for
tubes containing an anticoagulant to prevent clotting ■ Coagulation testing
of the sample. ○ Mix by inversion 3-4x only
● Anticoagulant prevent clotting by binding calcium or
inhibiting thrombin in the coagulation cascade
 Correct number of Inversions
Evacuated Tubes: Plasma (unclotted blood) Tubes
● Heparin
○ Dark green (non-gel) and light green (gel)
○ 2 anticoagulants
■ Sodium heparin
● Use for
○ Special chemistry tests
(Troponin assays),
Karyotyping
(Chromosomes) etc.
except electrolytes
● Fluoride
○ Grey top
○ 2 forms:
■ Sodium fluoride + Potassium oxalate
■ Sodium fluoride + K2 EDTA
○ Use for
■ Glucose testing
Order of Draw
○ Sample is stable as long as 24 hours at room
temperature
○ Ref: slow down metabolic process glycolysis
■ Glucose is broken down

● Black Top
○ Contain buffered sodium citrate
○ Used for
■ Westergren sedimentation rates.
○ Provide a ratio of blood to liquid anticoagulant
of 4:1
○ Specially designed tubes for westergren
sedimentation rates are available
○ ESR - erythrocyte sedimentation rate

● Yellow top
○ Sterile yellow stopper tubes
■ Contain the anticoagulant sodium
polyethanol sulfonate (SPS)
■ Use to collect samples to be cultured
for the presence of microorganisms
■ SPS prevents coagulation binding by
The order of draw recommended by CLSI for both
calcium
evacuated system and when filling tubes in a syringe:
■ SPS aids in the recovery of
1. Blood cultures
microorganisms by inhibiting the action
○ Yellow stopper tubes, culture bottles
of complement, phagocytes and certain
2. Light blue stopper tubes
antibiotics.
○ Sodium citrate
3. Red/gray, gold stopper tubes
○ Serum separator tubes
Red stopper plastic tubes
○ Clot activator
Red stopper glass tubes
4. Green stopper tubes and light green
○ Plasma separator tubes
○ Heparin
5. Lavender stopper tubes
○ EDTA
6. Gray stopper tubes
○ Potassium oxalate/ sodium fluoride
7. Yellow/gray or orange stopper tubes
○ Thrombin clot activator
Syringe Method ● Gloves w/ powder are not recommended because the
● Advantage powder can contaminate patient samples and cause
○ Phlebotomist is able to control the suction falsely elevated calcium values
pressure on the vein by slowly withdrawing the ● The glove powder can also cause a sensitization to
syringe plunger latex
○ Blood will appear in the hub of the needle
when the vein has been successfully entered Tourniquet
● Blood drawn in a syringe immediately transferred to ● Used during venipuncture to make it easier to locate
appropriate evacuated tubes to prevent the formation patients veins
of clots
● It is not acceptable to puncture the rubber stopper Puncture Site Protection Supplies
with the syringe needle and allow the blood to be ● 70% isopropyl alcohol
drawn into the tube. ○ Primary antiseptic used for cleansing the skin
in routine phlebotomy
Blood Transfer Device ● 2x2-inch gauze pads
● Provides a safe means for blood transfer without ○ Used for applying pressure to the puncture site
using the syringe needle or removing the tube stopper after the needle has been removed
● It is an evacuated tube holder with a rubber-sheathed ● Bandage or adhesive tape/microper
needle inside ○ Placed over the puncture site when the
● After blood collection, the syringe tip is inserted into bleeding has stopped
the hub of the device and evacuated tubes are filled ● It is not recommended to use cotton balls to apply
by pushing them onto the rubber-sheathed needle in pressure because the cotton fibers can stick to the
the holder as in an evacuated tube system. venipuncture site and may cause bleeding to begin
again when the cotton is removed.

Additional Supplies
● Clean glass slides
○ May be needed to prepare blood films for
certain hematology tests.
○ PBS - peripheral blood smear
● Pen/Marker
○ for labeling tubes

Winged Blood Collection Set

● “Butterflies”
● Used for
○ Infusion IV fluids
○ Performing venipuncture from very small or
very fragile veins often seen in children in the
geriatic population
● Usually
○ 21 or 23 gauge lengths of ½ to ¾ inch
● Plastic attachments to the needle that resemble
butterfly wings are used for holding the needle during
insertion.
● They also provide the ability to lower the needle
insertion angle when working with very small veins

Gloves
● Several varieties
○ Powdered
○ Powder-free
○ Latex
○ Non-latex
■ Vinyl
■ Nitrile
■ Neoprene
■ Polyethylene