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1- Blood-Borne Pathogens:

Infectious micro-organisms which live in


the bloodstream.
You can be exposed to bloodborne
pathogens if you are injured with a
contaminated needle.
You can also be exposed if your mucous
membranes, including eyes, mouth, or
the inside of your nose come into
contact with contaminated body fluids.

2- Personal Protective Equipment:


lab coat
Gloves
Face masks

3- Hand Washing:
Hand washing is the single most important infection
control measure.
Wash hands thoroughly before, after, and between
all patient contacts.
Be sure to turn off faucets using a paper towel to
avoid contamination.
Remove rings
Stand by the sink but do not touch it
Apply soap and rub hands together
Both sides of the hand, between fingers, around
knuckles, under fingernails
Rinse hands in a downward motion
Dry hands with a clean paper towel
Turn off water with another paper towel

4- Hazardous waste disposal:


All needles & other sharps must be
disposed of in approved sharps disposal
containers.
Other contaminated waste must be
discarded in an appropriate biohazard
bag or waste receptacle.
5- Needle stick:
Safety Devices should always be
encouraged when handling needles
and sharps

Whole blood

A venous, arterial or capillary blood sample in


which the concentrations and properties of
cellular and extra-cellular constituents
remain relatively unaltered when
compared with their in-vivo state.
Anticoagulation in-vitro stabilizes the
constituents in a whole blood sample for a
certain period of time.

Plasma

The virtually cell-free supernatant of blood


containing
anticoagulant obtained after
centrifugation.

Serum
The undiluted, extracellular portion of blood
after adequate coagulation is complete.

Basic Steps for Capillary Puncture


1. Patient identification, isolation and dietary restrictions
2. Reassure the patient
3. Position the patient
4. Assemble the supplies needed
5. Verify the tests requested
6. Choose the puncture site
7. If necessary, warm the site
8. Cleanse the area
9. Perform the skin puncture
10. Wipe the first drop
11. Collect the blood sample into the appropriate collection devices
12. Cap or seal the collection device
13. Apply pressure to site
14. Discard the lancet
15. Label the specimens and check for proper sample handling
16. Date, sign, and record time on sample and paperwork
17. Document on requisition that the sample was collected by a skin
puncture
18. Transport specimens properly to the laboratory

Steps for Venipuncture Procedure


1. Patient identification, isolation and dietary restrictions
2. Reassure the patient
3. Position the patient
4. Assemble the supplies needed
5. Apply the tourniquet and choose the phlebotomy site
6. Cleanse the area
7. Perform the venipuncture
8. Release the tourniquet
9. Remove the needle and position the gauze
10. Apply pressure
11. Fill tubes if syringe is used
12. Dispose of needle unit
13. Label specimen and check for proper sample handling
14. Date, sign, and record time on sample and paperwork
15. Transport tubes properly to laboratory

RBCs, Hb, Ht: in venous blood (lower in


capillary bl.)
(Hypoxia of venous blood RBCs)
Platelets count: in venous blood (lower
in capillary bl.)

A plastic holder must be used with the


evacuated tube system.

Needle holders with built-in protection


devices

Single draw needles


Multiple Draw Needle
Butterfly Needle

Anticoagulants - Advantages &


Disadvantages

Heparin

Heparin is a mucoitin polysulfuric acid that is


available as sodium, potassium and ammonia
salts. It acts as an antithrombin which prevents
the transformation of prothrombin into thrombin,
which in turn, prevents the formation of fibrin
from fibrinogen. Normally it takes about 20 units
of heparin to anticoagulate 1 ml of blood.

Heparin
Advantages:
1. causes the least interference with tests
2. it can be used when testing for
phosphorous
3. heparin is available in liquid or powder
form and dissolves quickly

Heparin

Disadvantages:
1. heparin is expensive.
2. the action is temporary.
3. it produces a blue background in blood
smears stained with Wright's stain.
4. acid phosphatase activity is inhibited.
5. in certain calcium methods heparin can
interfere with the binding of calcium.

EDTA (Ethylenediaminetetraacetic acid)

EDTA prevents coagulation by chelating or binding


the calcium in the blood. The concentration required
for effective anticoagulation is 1-2 mg/mL of blood.
EDTA is the most commonly used anticoagulant used
in hematologic studies because it preserves the
cellular components. It also has little effect on other

clinical chemistry tests (except for those listed below).


If EDTA : platelets, MCHC
heamatocrite

EDTA (Ethylenediaminetetraacetic acid)


Advantages:
1. Best for CBC:

- Preserves morphology
- No platelets clumping
- ESR not altered if
blood is refrigerated

EDTA (Ethylenediaminetetraacetic acid)


Disadvantages:
1. EDTA inhibits alkaline phosphatase, CK,
and leucine aminopeptidase activities
through chelation of the metallic
cofactors.
2. Because EDTA chelates calcium, it is not
suitable for calcium and iron analysis.

Sodium Fluoride
Sodium

fluoride

acts

as

weak

anticoagulant and is usually considered


the preservative of choice for blood
glucose levels. It exerts its preservative
action by inhibiting the enzyme involved
in glycolysis.

Sodium Fluoride

Disadvantages:
1. Fluoride acts as an inhibitor on serum
enzymes and in increased concents
affects urease, which is used in measuring
BUN.
2. There is little justification for its use as an
anticoagulant for clinical chemistry tests.
3. To be used as an anticoagulant, increased
concentration/ml would be needed. The
increased concentrations and inhibition of
the glycolytic cycles would cause fluid
shifts.

Citrate
Citrate is used as Sodium citrate and is mixed
1 part to 9 parts of blood. Na citrate is most
commonly used in coagulation studies.
Calcium is chelated and thus the effect
can be reversed by the addition of ionized
calcium.

Citrate
Uses:
1. ESR: 4 volumes of blood + 1 volume of
citrate
2. Coagulation tests: 9 volume of blood + 1
volume of citrate

Citrate

Disadvantages:
1. Na citrate has little application in chemistry
tests.
2. Since the citrate chelates calcium, it is
unsuitable in measurement of this
substance.
3. It inhibits aminotransferases and alkaline
phosphatase.
4. Citrate also complexes with molybdate
(used in the analytical procedure for
phosphorous).

Oxalates
Sodium, potassium, ammonia, and lithium
oxalates are available with potassium
oxalate being the most widely used. Acts
by forming an insoluble complex with the
calcium ions. It is used in the concentration
of about 1-2 mg/mL.

Oxalates
Disadvantages:
1. Na and Li oxalates can cause shrinkage of
RBC by drawing water into the plasma.
2. Oxalates can cause a 10% reduction in the
hematocrit and a 5% reduction in the
concentration of plasma constituents.
3. Oxalates inhibit the enzymes acid and
alkaline phosphatase, amylase, and LDH.

sodium citrate.
coagulation (clotting) studies.
must be completely filled
must be inverted immediately after filling

sodium or lithium heparin


for tests requiring whole blood or plasma
such as ammonia

heparin or Na EDTA anticoagulants


Tube is designed to contain no
contaminating metals
Trace element and toxicology studies

Inhibitor for glycolysis + anticoagulant


Sodium Fluride +potassium oxalate.
glucose levels.

EDTA to prevent clotting


hematology studies.
Should be completely filled
Must be inverted after filling

No additives
Blood bank tests, toxicology, serology
Must not be inverted after filing

Acid citrate dextrose


Inactivates complements
DNA studies, paternity testing

Contains tri sodium citrate.


Used for ESR also known as Erythrocyte
Sedimentation Rate

Containers containing coagulants

Gold or 'Tiger' Red/Black top: Clot activator and gel for serum
separation

Orange or Grey/Yellow 'Tiger' Top: Contain Thrombin, a rapid


clot activator, for STAT serum testing (Short Turn Around Time)

< 1 hour: for PT & PTT (coagulation profile), if not to


be done immediately, separate plasma & freeze
3 hours: for CBC, ESR, Ht, indices, fragility test
>3 hours: - RBCs start to swell
- WBCs and platelets start to fall
6 hours: Reticulocytes disappear
24 hours: Normoblasts disappear
1-2 days: RBCs, WBCs, Hb done within 1-2 days if
there is a problem (better fresh within 3 hs)
4 hours: bilirubin is done before that time (for pt
with hemolytic anemia)
NB. Blood films must be done from last drop in
syringe or finger prick or within 1 hour from blood
on EDTA.