1- Blood-Borne Pathogens: Infectious micro-organisms which live in

the bloodstream. You can be exposed to bloodborne

pathogens if you are injured with acontaminated needle.

You can also be exposed if your mucousmembranes, including eyes, mouth, orthe inside of your nose come intocontact with contaminated body fluids.

1- Blood-Borne Pathogens: Infectious micro-organisms which live in

the bloodstream. You can be exposed to bloodborne

pathogens if you are injured with acontaminated needle.

You can also be exposed if your mucousmembranes, including eyes, mouth, orthe inside of your nose come intocontact 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

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 disposalcontainers.

Other contaminated waste must bediscarded in an appropriate biohazardbag or waste receptacle.

5- Needle stick: Safety Devices should always be

encouraged when handling needlesand sharps

4- Hazardous waste disposal: All needles & other sharps must be

disposed of in approved sharps disposalcontainers.

Other contaminated waste must bediscarded in an appropriate biohazardbag or waste receptacle.

5- Needle stick: Safety Devices should always be

encouraged when handling needlesand sharps

Whole bloodA venous, arterial or capillary blood sample in

which the concentrations and properties ofcellular and extra-cellular constituentsremain relatively unaltered whencompared with their in-vivo state.Anticoagulation in-vitro stabilizes theconstituents in a whole blood sample for acertain period of time.

Whole bloodA venous, arterial or capillary blood sample in

which the concentrations and properties ofcellular and extra-cellular constituentsremain relatively unaltered whencompared with their in-vivo state.Anticoagulation in-vitro stabilizes theconstituents in a whole blood sample for acertain period of time.

Plasma The virtually cell-free supernatant of blood

containing anticoagulant obtained after

centrifugation.

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 restrictions2. Reassure the patient3. Position the patient4. Assemble the supplies needed5. Verify the tests requested6. Choose the puncture site7. If necessary, warm the site8. Cleanse the area9. Perform the skin puncture10. Wipe the first drop11. Collect the blood sample into the appropriate collection devices12. Cap or seal the collection device13. Apply pressure to site14. Discard the lancet15. Label the specimens and check for proper sample handling16. Date, sign, and record time on sample and paperwork17. Document on requisition that the sample was collected by a skin

puncture18. Transport specimens properly to the laboratory

Basic Steps for Capillary Puncture

1. Patient identification, isolation and dietary restrictions2. Reassure the patient3. Position the patient4. Assemble the supplies needed5. Verify the tests requested6. Choose the puncture site7. If necessary, warm the site8. Cleanse the area9. Perform the skin puncture10. Wipe the first drop11. Collect the blood sample into the appropriate collection devices12. Cap or seal the collection device13. Apply pressure to site14. Discard the lancet15. Label the specimens and check for proper sample handling16. Date, sign, and record time on sample and paperwork17. Document on requisition that the sample was collected by a skin

puncture18. Transport specimens properly to the laboratory

Steps for Venipuncture Procedure

1. Patient identification, isolation and dietary restrictions2. Reassure the patient3. Position the patient4. Assemble the supplies needed5. Apply the tourniquet and choose the phlebotomy site6. Cleanse the area7. Perform the venipuncture8. Release the tourniquet9. Remove the needle and position the gauze10. Apply pressure11. Fill tubes if syringe is used12. Dispose of needle unit13. Label specimen and check for proper sample handling14. Date, sign, and record time on sample and paperwork15. Transport tubes properly to laboratory

Steps for Venipuncture Procedure

1. Patient identification, isolation and dietary restrictions2. Reassure the patient3. Position the patient4. Assemble the supplies needed5. Apply the tourniquet and choose the phlebotomy site6. Cleanse the area7. Perform the venipuncture8. Release the tourniquet9. Remove the needle and position the gauze10. Apply pressure11. Fill tubes if syringe is used12. Dispose of needle unit13. Label specimen and check for proper sample handling14. Date, sign, and record time on sample and paperwork15. Transport tubes properly to laboratory

RBCs, Hb, Ht: ↑ in venous blood (lower incapillary bl.)(Hypoxia of venous blood→↑ RBCs)

Platelets count: ↑ in venous blood (lowerin capillary bl.)

RBCs, Hb, Ht: ↑ in venous blood (lower incapillary bl.)(Hypoxia of venous blood→↑ RBCs)

Platelets count: ↑ in venous blood (lowerin capillary bl.)

A plastic holder must be used with theevacuated tube system.

Needle holders with built-in protectiondevices

Single draw needles Multiple Draw Needle Butterfly Needle

Anticoagulants - Advantages &Disadvantages

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

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.

HeparinAdvantages:1. causes the least interference with tests2. it can be used when testing for

phosphorous3. heparin is available in liquid or powder

form and dissolves quickly

HeparinAdvantages:1. causes the least interference with tests2. it can be used when testing for

phosphorous3. heparin is available in liquid or powder

form and dissolves quickly

HeparinDisadvantages: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.

HeparinDisadvantages: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) 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 ifblood is refrigerated

EDTA (Ethylenediaminetetraacetic acid)Advantages:1. Best for CBC: - Preserves morphology

- No platelets clumping- ESR not altered ifblood is refrigerated

EDTA (Ethylenediaminetetraacetic acid)Disadvantages:1. EDTA inhibits alkaline phosphatase, CK,

and leucine aminopeptidase activitiesthrough chelation of the metalliccofactors.

2. Because EDTA chelates calcium, it is notsuitable for calcium and iron analysis.

EDTA (Ethylenediaminetetraacetic acid)Disadvantages:1. EDTA inhibits alkaline phosphatase, CK,

and leucine aminopeptidase activitiesthrough chelation of the metalliccofactors.

2. Because EDTA chelates calcium, it is notsuitable for calcium and iron analysis.

Sodium FluorideSodium fluoride acts as a 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 FluorideSodium fluoride acts as a 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 FluorideDisadvantages:1. Fluoride acts as an inhibitor on serum

enzymes and in increased concentsaffects urease, which is used in measuringBUN.

2. There is little justification for its use as ananticoagulant for clinical chemistry tests.

3. To be used as an anticoagulant, increasedconcentration/ml would be needed. Theincreased concentrations and inhibition ofthe glycolytic cycles would cause fluidshifts.

Sodium FluorideDisadvantages:1. Fluoride acts as an inhibitor on serum

enzymes and in increased concentsaffects urease, which is used in measuringBUN.

2. There is little justification for its use as ananticoagulant for clinical chemistry tests.

3. To be used as an anticoagulant, increasedconcentration/ml would be needed. Theincreased concentrations and inhibition ofthe glycolytic cycles would cause fluidshifts.

Citrate

Citrate is used as Sodium citrate and is mixed1 part to 9 parts of blood. Na citrate is mostcommonly used in coagulation studies.Calcium is chelated and thus the effectcan be reversed by the addition of ionizedcalcium.

Citrate

Citrate is used as Sodium citrate and is mixed1 part to 9 parts of blood. Na citrate is mostcommonly used in coagulation studies.Calcium is chelated and thus the effectcan be reversed by the addition of ionizedcalcium.

CitrateUses:1. ESR: 4 volumes of blood + 1 volume of

citrate2. Coagulation tests: 9 volume of blood + 1

volume of citrate

CitrateUses:1. ESR: 4 volumes of blood + 1 volume of

citrate2. Coagulation tests: 9 volume of blood + 1

volume of citrate

CitrateDisadvantages:1. Na citrate has little application in chemistry

tests.2. Since the citrate chelates calcium, it is

unsuitable in measurement of thissubstance.

3. It inhibits aminotransferases and alkalinephosphatase.

4. Citrate also complexes with molybdate(used in the analytical procedure forphosphorous).

CitrateDisadvantages:1. Na citrate has little application in chemistry

tests.2. Since the citrate chelates calcium, it is

unsuitable in measurement of thissubstance.

3. It inhibits aminotransferases and alkalinephosphatase.

4. Citrate also complexes with molybdate(used in the analytical procedure forphosphorous).

Oxalates

Sodium, potassium, ammonia, and lithiumoxalates are available with potassiumoxalate being the most widely used. Actsby forming an insoluble complex with thecalcium ions. It is used in the concentrationof about 1-2 mg/mL.

Oxalates

Sodium, potassium, ammonia, and lithiumoxalates are available with potassiumoxalate being the most widely used. Actsby forming an insoluble complex with thecalcium ions. It is used in the concentrationof about 1-2 mg/mL.

OxalatesDisadvantages: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 theconcentration of plasma constituents.

3. Oxalates inhibit the enzymes acid andalkaline phosphatase, amylase, and LDH.

OxalatesDisadvantages: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 theconcentration of plasma constituents.

3. Oxalates inhibit the enzymes acid andalkaline phosphatase, amylase, and LDH.

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

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

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

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)

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 tobe 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 bloodon EDTA.

< 1 hour: for PT & PTT (coagulation profile), if not tobe 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 bloodon EDTA.