MLS REVIEWER FOR MIDTERM (LABORATORY)
ACTIVITY 5 BLOOD SPECIMEN COLLECTION BY
VENIPUNCTURE
 The TESTING OF LABORATORY SPECIMENS are
essential to the correct diagnosis, treatment and
monitoring of the condition of a patient.
 70% of gathered data from the laboratory results are
used by health physicians to determine the disease
or illness of a patient.
 Inaccurate results and poor findings can lead to
the wrong treatment of a patient that will result to
under-medication, over-medication and even death
as the worst-case scenario.
 Finding a test approx. 56% of misreading in
laboratories occur during the pre-analytical phase or
before samples are subjected to tests.
SAFETY PRECAUTIONS:
Blood borne-pathogens are:
 Human immunodeficiency virus (HIV)
 Hepatitis B virus (HBV)
 Hepatitis C virus (HCV).
The following are the standard precautions that must be
observed by the medical personnel in patient care:
1. Abiding by isolation practices
2. Wearing proper personnel protective equipment (PPE)
3. Observing proper hygiene
 Alcohol solutions may only be used as an
alternative for hand washing if the hands are
not evidently contaminated, the phlebotomist
did not enter or stay in an isolation room with a
patient infected with highly motile bacteria such
as Clostridium difficile.
4. Properly using safety device
5. Placing collection tray closely
6. Correctly packaging specimens
7. Labeling and cushioning specimens for transport
8. Reporting all accidental exposure to needle sticks and
starting post-exposure prophylaxis (PEP) when needed
9. Disposing entire assemblies including tube holders and
needles after use
10. Disposing contaminated materials in designated
containers
11. Decontaminating surfaces
 Surfaces exposed to potentially hazardous and
infectious matters should be decontaminated with
disinfectants such as SODIUM HYPOCHLORITE
( diluted at 1:10 or 1:100 proportion for routine
decontamination)
 Personnel who work in off-site facilities or physician’s
office may be required to perform initial work on
specimens such as CENTRIFUGATION and
SERUM or PLASMA SEPARATION.
LESSONS IN ACTIVITIES 5.1 - 5.4:
 VENIPUNCTURE is defined as the process of
collecting or drawing blood from a vein.
 There are two ways by which blood can
be collected by venipuncture:
 The first is using the needle and syringe
which is also referred to as the open
system. And the second is through
evacuated tube system referred to as
the closed system.
 Obtain and accession the test request:
(a) manual requisitions, (2) computer generated
requisitions, (3) barcode labels
 Explain the procedure to enlighten them, before
dealing with patients wash your hands first to avoid
contamination of diseases or viruses.
 Then, in doing venipuncture to the patients. Ask
them first their personal data like name, age and
birthdate.
 Patients arm should be supported firmly and arm
should not be bent at the elbow.
 Supine patients should be arm extended
and not bent at elbow.
 The patient’s hands should be fist. Do not
allow the patient to pump (open and close)
fist as this causes Hemoconcentration
 In selecting venipuncture site it should be bouncy  In verifying the test results, there will a signature
and resilience. Use the tip of the index finger to from a phlebotomist or medical technologist,
palpitate the veins. validator and physician.

 Do not draw from above an IV if

necessary, have IV turned off for a
minimum of 2 minutes. SPECIAL TEST THAT REQUIRES PROCEDURE TO
FOLLOW:
 Avoid burned, scarred or tattooed
areas because it was already Putting on crushed ice (Chilled Specimen)
contaminated.
Lab notes:
 Avoid edematous areas
 Ammonia
 Avoid drawing in the area of Hematoma,
draw from below the site if there is no  lactic acid
other alternative.
 blood gases
 Mastectomy patients, draw from the side
 B-natriuretic peptide
opposite of mastectomy whenever
possible. If patient has double  ABG
mastectomy, the physician should be
consulted usually to avoid the side of the Lecture notes:
most recent mastectomy.
 adrenocorticotropic hormone (ACTH)
 Use leg, ankle, or foot veins as a last
resort and after obtaining permission from  acetone, angiotensin-converting enzyme (ACE)
the patient’s physician.
 ammonia, catecholamines
 Lower extremity venipuncture may cause
blood clot formation.  free fatty acids

 Use with antiseptic (70% isopropyl alcohol). Use  Gastrin

circular motion cleaning the anticubital fossa area.
Do not touch the area that already clean to avoid  Glucagon
contamination.
 Homocysteine
 Applying tourniquet should be less than or exact 1
 lactic acid
minute then remove it. More than 1 minute can
cause Hemoconcentration.
 parathyroid hormone (PTH)
 In anchoring vein it should be 1 to 2 inches in
 ph/blood gas (if indicated)
placing your thumb. Have the patient make fist and
line up the needle bevel up in the direction of the  Pyruvate
vein.
 renin
 Insert the needle at 15 degrees to 30 degrees.
Keeping it in warm (Body Temperature)
 After doing venipuncture, apply pressure using
gauze on the site. Lab notes:

 Put the details that are needed in order to easily  cold agglutinin
distinguish a patient.
 Cryoglobulin

 Cryofibrinogen
Protecting from light (Light-sensitive Specimen) ACTIVITY 8 BLOOD COLLECTION FOR CULTURE

Lab notes:
 BLOOD CULTURE - are ordered by a physician
when there is FEVER OF UNKNOWN ORIGIN (FUO)
 Bilirubin or a reason to suspect that there is BACTEREMIA
(bacteria in the blood) or SEPTICEMIA
Lecture notes: (pathogenic bacteria in the blood)

 Carotene
 ANAEROBIC CULTURE BOTTLE- allowing
preferential growth of strict anaerobic bacteria.
 Red cell folate
 The specimens included here are: blood
 Serum folate specimens, aspirates of body
fluids (pleural, pericardial,
 Vitamin B2 cerebrospinal, peritoneal, and joint
fluids), urine collected by suprapubic
 Vitamin B6 aspiration, abscess contents, deep
wound aspirates, and specimens
 Vitamin B12 obtained by special procedures such as
transtracheal.
 Vitamin C
 Growth Medium :
 Urine porphyrins
Thioglycollate, SPS
 Urine porphobilinogen
 * 5-10 mL of blood sample

 AEROBIC BLOOD CULTURE BOTTLE- a one

ANTISEPTICS: bottle is designed to enhance the growth of aerobic
organisms.
 70% Isopropyl Alcohol
 The samples here are: Nocardia sp,
 70% Ethyl Alcohol Pseudomonas aeroginusa, E. coli,
citrobacter, klebsiella, proteus,
 Povidone-Iodine salmonella, achromobacter
mycobacterium tuberculosis.
 Tincture Iodine
 Growth Medium:
 Benzalkonium Chloride
Tryticase soy Broth, Resin, SPS
 Chlorhexidine Gluconate
 * 5-10 ml
 Hydrogen Peroxide
 Providone iodine and benzalkonium chloride are
the antiseptic use for blood culturing.

ADDITIONAL NOTES:  Blood collection bottles (aerobic, anaerobic, ARD,

or tubes)
 Analytes:
Procedures:
 Proteins
1. Clean the site with antiseptic 70% isopropyl
 Irons alcohol for a minimum of 30 seconds.

 Cholesterol 2. Use a tincture of iodine for 30 seconds or a

providone iodine swabstick for 60 seconds to
 Lipid cleans the site.

 Red tube - is prone to hemolyze

3. Remove the protective flip top covering the
rubber septum and clean the tops of the bottles
with 70% alcohol or iodine.
4. Mark minimum and maximum fill lines. Adult: 5-10
Ml bottle. Note the several different ways or means
by which broth inoculation can occur. Collect directly
into the bottles using a butterfly syringe needle.
5. Fill the aerobic vial first. Avoid backflow by
keeping the culture bottle lower than the collection
site. Fill anaerobic and aerobic bottles with 5-10
mL of blood.
6. Fill pediatric bottles with 1-3 mL of blood.
Blood culture bottles top colors:
 YELLOW- aerobic pedia
 GREEN- aerobic adult 8 – 10 inversions
 ORANGE- anaerobic
Syringe method:
1. Transfer the blood into the bottles after the draw is
completed. Use a transfer device. Fill anaerobic vial
first.
2. If using the needle on the syringe for transfer, do not
hold the culture bottle with your hand to avoid
accidental needlesticks.
3. Intermediate collection tube. A sodium polyanethol
sulfonate (SPS), yellow top is acceptable. Blood
must be transferred from the tube into the culture
bottles.
4. Check the patients arm and clean the iodine from
the patient’s skin with an alcohol pad.
5. Leading culture bottles including the site of collection.
Dispose of contaminated materials. Thank the
patient.
6. Remove gloves, check the specimen collection logs
and transport the specimens to the laboratory.
NOTES:
 Collection of Blood Cultures in a timely fashion is
important. Commonly, blood cultures are ordered
before or immediately after fever spikes.
 Skin antisepsis is the most important part of the
blood culture.
ACTVITY 12: PREPARATION OF SERUM OR PLASMA
FOR LABORATOTORY TESTING
 PHLEBOTOMISTS are not only required to draw
patient blood specimens, but are also expected to
process the specimen. Specimen processing
involves centrifuging the patient’s blood and
possibly, urine samples.
 CENTRIFUGATION is a process of spinning down
or separating the cells from the liquid portion of the
blood. Centrifuges come in various styles:
 Refrigerated
 Floor models
 Table top models.
 The speed of rotation (revolutions per minute, or
rpm) and the radius of the rotor head determine
the relative centrifugal force (RCF) of a
centrifuge.
 Centrifugal force expressed as gravity(g). Most
laboratory specimens are centrifuged for 15
minutes at 750 to 1000g.
 Centrifuges are intended to spin bloo specimens
down, separating the cells from the liquid portion.
 Different tubes can be spun in the same centrifuge
at the same time. Make sure to put tubes of
equivalent size and volume specifically opposite one
another, in short, always check the levels of the
sample so that the centrifuge will be balanced. In
case you do not have an even number of blood
tubes to spin, you can balance centrifuge with
similar tubes filled with water or saline.
 If the centrifuge is loaded of samples it will make
vibrate or noise. The centrifuge must be calibrated
on a regular schedule with a TACHOMETER to
ensure proper centrifugation.
 Improper use of centrifuge can be dangerous to the
user and can damage laboratory specimens.
 Never open a centrifuge lid until the rotor has come
to a complete stop. If you open the lid before it
stops, you could be at risk of being injured by the
flying shards or debris
 ALIQUOTING is the method of dividing or
separating specimens into separate containers. For
most laboratory tests require serum or plasma, it is
suggested that specimens be separated within
two hours of collection or less.
 If samples are not centrifuged or aliquoted within two
hours, the laboratory test results might be altered.
 If the red blood cells are left in contact with the 15. Take great care not to put the wrong specimen in
serum or plasma, glucose would be decreased and the wrong tube.
potassium could significantly increase because cells
use glucose to keep themselves alive and
nourished. This is known as GLYCOLYSIS.

 Potassium and glucose blood tests are most

affected, as well as coagulation studies, because
the blood is allowed to sit for longer than two hours.

Materials:
 Centrifuge

 Test tube with 3 mL NSS

 Test tube with 3 mL whole blood with proper

label

 Transfer test tube with a label, same as the

label of the test tube for whole blood

Procedure:

1. Transport specimen to the centrifuge area.

2. Re-check if the test tubes are properly labeled.

3. Open the lid of the centrifuge

4. Conveniently load the test tubes opposite each other

in the centrifuge.

5. Ensure that the tubes are balanced

6. Check if all the tubes have covers. If there is a tube

with a missing cap, cover the end of the tube.

7. Close and lock the centrifuge lid.

8. Set the centrifuge time and speed correctly.

9. Allow the centrifuge to stop completely.

10. Open the centrifuge lid only after the rotor has
completely stopped.

11. Remove the tubes carefully.

12. Wipe down the rotor and centrifuge after use.

13. Transfer the liquid serum/plasma into another test

tube using the Pasteur pipette.

14. Label the transfer tube in which the aliquot of serum

or plasma is placed before the aliquot is added to
the tube.