 Specimen/ samples – analyzed  antecubital fossa veins – median (most preferred), cephalic, ADVANTAGES OF VENIPUNCTURE OVER SKIN PUNCTURE:

 Substances – measured basilica  Large amount of blood can be obtained for a variety of
SUBSTANCES MEASURED IN SERUM  wrist, hand & ankle veins tests
1. Substances normally present w/ function: Complications (immediate/ delayed & local/ systemic):  Additional & repeated test can be done
- glucose, TP, albumin, individual proteins, electrolytes,  hematoma (missed vein)  Fastest method of collecting samples from a number of
total albumin globulin (TAG), cholesterol, hormones, vit  collapsed small veins (excessive pull of plunger) patients
2. Metabolites – waste-products  syncope (loss of consciousness)  Blood can be transported to lab and stored for future
- urea, creatinine, uric acid, NH3, Bb  excessive bleeding use
SAMPLE VARIABLES:  thrombosis of vein  blood chemistry determination
1. Physiological consideration  blood-borne infection: Hep B & AIDS DISADVANTAGES OF VENIPUNCTURE:
2. Proper Px preparation Methods:  Harm on infants, children & obese individuals
3. Problems w/ collection a. Syringe method (most conventional)  more time & skill (operator)
4. transport - single sample needles – specifically for syringes  complications
5. processing - barrel, plunger, needle, needle holder (5&10), hub, bevel COMPLICATIONS IN VENIPUNCTURE:
6. storage b. Evacuated tube system 1. local immediate complications
IMPORTANT CONSIDERATIONS - multiple needles should be used if more than 1 tube of - hemoconcentration
1. Physical activity blood is to be collected (prevent leakage during tube - failure of blood to enter syringe
a. Transient effects – inc, dec (FFA, alanine, lactate, K+) changes - circulatory failure
b. Long term effects – inc alanine, aldolase, ALT, LDH - gel: barrier bet serum & cells - fainting or syncope
c. 6 months of physical training – inc: - marked w/ expiration date (lost vacuum/ ineffective 2. local delayed complications
- plasma testosterone: 21% additive) - hematoma
- androstenedione: 25% - correct blood-AC ratio - thrombosis of vein
- LH: 25% - needle: 2 way longer for tube; other for vein - thrombophlebitis
2. Fasting (prolonged)—FBS: 12hrs c. Butterfly infusion set 3. general delayed complications
a. after 48 hrs: serum Bb inc: 240% - stainless steel beveled needle w/ attached plastic wing for - serum hepatitis, AIDS
b. after 72 hrs: glucose dec to 45mg/dl phlebo to grasp during needle insertion  SST – clot activator – reduce clotting time
- inc plasma trig, glycerol & FFA - plastic tubing connects needle to adaptor- screws into a  PST – green/ gray
- cholesterol: no change tube holder = modified evacuated system - Li Heparin w/ gel
3. Posture - for pedia/ babies & geriatrics - Electrolytes, routine chem. (invert 505x)
- supine upright (inc albumin, drug-bound CHON, *gauge: bigger number = smaller bore (21:ideal; 26: blood  Trace element tube: heparin, EDTA or none
cholesterol, enzymes, total CHON, triG, Bb, Ca) donation)  SPS: Soidum/ Polyanithol sulfonate (micro)
4. Ethanol ingestion VENIPUNCTURE PROCEDURE  ACD – acid citrate dextrose (HLA typing)
- inc plasma lactate, urate, trig 1. assembling the patient PARTS:
- ROH abusers – inc GGT (γ-glutamyl transferase), urate, 2. positioning & ID the patient 1. needles
MCV 3. preparing needle & evacuated tubes or syringe - adults: 20, 21
5. Tobacco smoking 4. applying tourniquet to select the vein & removal of - pediatrics/ geriatrics: 22; veterinary: 18
- inc blood carboxy Hb levels tourniquet 2. needle holder
a. heavy smokers: inc 8% - 3 mins application increases CHON, Fe, AST, Bb & total - regular – 13, 16mm
b. non-smokers - <1% lipids - pediatric – 10mm
c. acute tobacco smoking – inc plasma catecholamines, - Repeated fist clinching increases K+ 1-2nm 3. tubes/ evacuated tubes
serum cortisol 5. applying the antiseptic (wet then dry) - regular – 75mm; 100mm
GENERAL RULES FOR BLOOD SPECIMEN COLLECTION: - circular motion (inside to out) ARTERIAL PUNCTURE
1. Px ID 6. reapplying tourniquet - oxygenated blood; uniform composition throughout body
2. approaching Px 7. inserting the needle - measure O2 , CO2 tension & blood pH
3. Collection site 8. withdrawing blood - BGA: critical to patients w/ pulmonary problems, O2
4. Tourniquet app - IV avoided therapy, cardiovascular problems & those undergoing
5. checking Px - Turn off IV for 2-5 mins, discard 1st 5ml of draw major operations
6. specimen handling & transport 9. releasing tourniquet - Sites: radial, brachial & femoral arteries
7. storing specimen prior to transport 10. withdrawing needle - Radial & brachial: preferred sites
 serum pH: 8.5 – ACP destroyed 11. removing needle from syringe/ removing evacuated - Newborns: umbilical artery catheter
 Glycolysis – dec serum glucose tube from needle - Brachial: 18-20 gauge, 45-60
 Changes in RBC permeability – inc K, P, Mg 12. transferring blood to an appropriate container - Radial: 23-25, 90
 Freezing, subseq thawing, refreezing, rethawing: CHON 13. checking patient’s wound - AC: heparin
denaturation - Betadine – falsely high P, uric acid, K+ Complications:
 Frozen specimen, thawed at RT before analysis – inc ALP - Isopropyl (70%) alcohol should not be used for medical or  Hematoma (increases pressure in artery)
 Exposure to light – dec Bb legal ethanol levels  Arterial spasm (restriction of BF due to reflex
VENIPUNCTURE 14. labeling specimen constriction)
- deoxygenated blood; contains subs that come from ADVANTAGES OF EVACUATED TUBE SYSTEM OVER SYRINGE  Temporary discomfort (aching, throbbing, tenderness,
metabolic activities of diff organs METHOD: sharp sensation & cramping)
- blood chem. & immunologic studies  multiple blood collection  Thrombosis, hemorrhage & infection
- more easily collected than arterial blood  no prior preparation Considerations:
Sites:  wider range of tube sizes & AC  Intense care must be administered
 safer method of blood collection
  Not be selected: irritated, edematous, near wound or area  avoid pressure & squeezing e acts as AC
of AV shunt or fistula  first drop should be discarded (alcohol & tissue juice)  Inhibits
 Ice water/ coolant (1-5C): minimize WBC O2  RBC, Hct, Hb & platelets: low glyceraldeh
consumption  WBC: higher than in venous ydes 3-
 Capillary blood: substitute for arterial blood determination  Capillary blood from skin puncture is an admixture of phosphate
of pH & pCO2 (warmed site: increases BF through venous, arterial & may contain tissue juice dehydrogen
capillaries & arterioles arterial-rich blood) Microcapillary tubes used in hema: ase
SKIN PUNCTURE/ CAPILLARY PUNCTURE  Blue ring – no AC Yellow Citrate Preserves Blood culture
- small quantity blood needed  Red ring – heparinized dextrose RBC
- pediatrics, obese w/ thrombotic tendencies & severe  Green ring – heparinized Green Heparin (Na+. Inhibits Active &
burns  Skin puncture site must be warmed (increase BF): Li+ or NH4+) thrombin methemoglob
- geriatric patients: thinner & less elasticity of skin  Dry heat or paper towel w/ warm H2O (39-42C) BGA, in
Sites:  Flicking w/ index finger until flushing is observed ammonia
 earlobe – free edges, not on sides  Chemicals (trafutil paste Histamine creams) CO-Hb
 palmar surface of finger  Arterial blood from capillary puncture may yield unreliable  Anticoagulant interference:
 plantar surface of heel & big toe results if:  Dilution errors—oxalates: highly osmotic
Infants:  Systolic is less than 95mmHg  Inhibition of plasma enzyme activities—Fluoride
 digits of 2nd, 3rd or 4th fingers  Cardiac output is severely restricted (enzyme poison)
 lateral plantar heel surface  Vasoconstriction - EDTA: chelates metallic enzyme activators
 median plantar heel surface  Puncture depth: 0.85-2mm deep; 1.75-3mm length - Oxalates: inhibits AMS, LD & ACP
Children VACUTAIJNER/ EVACUATED TUBES: - Citrate: inhibits AMS
 plantar surface of big toe Color coded based on AC (2,5,7 &10ml)  Oxalates, citrate & EDTA: lower plasma Ca levels
 lateral side of finger, adjacent to nose 1. Yellow (blood culture tubes)  False increase in electrolyte analyses due to AC in salt
Blood obtained: 2. Red (no AC—obtain serum) form
 capillary: BG (prewarmed) 3. Blue (Na citrate) SPECIMEN HANDLING & PROCESSING
 peripheral (most common term used) 4. Green (heparin) Serum:
 arteriolar 5. Lavender (EDTA)  20-30 mins: ideal clotting time
Sites to avoid: 6. Others--Gray (Fluoride-glucose det)  Preferred than plasma:
 inflamed or palmar surface  Gel vs non gel: serum or plasma separator tubes maybe 1. interfering substance are co-precipitate during clotting,LPL
 congested or edematous site unacceptable for some analytes (therapeutic drug) 2. optically clearer
 cold & cyanotic areas (concentrated blood) Color Additive Action Use 3. free from AC interference
 scarred & heavily calloused area Lavende EDTA Chelates Hematologic  Reach lab w/in 45 mins
Advantages using earlobe: r  Versene calcium assays, lead  Agitation avoided
 less painful (few nerve endings) (disodium assays, CEA  Amber containers for photolabile substances
 free flow of blood (thinner skin) salt) determination  Transport in ice (4C): BGA, rennin, enzymes &
 less tissue contamination of blood  Sequestren & cell counts catecholamines
 searching Abnormal cells (histiocytes in bacterial e Specimen Interference:
endocarditis) (dipottasium  Lysis of cells (hemolyzed serum)
Advantages using finger: salt)  Leakage if IC substances
 accessible to phleb Red none Allow blood Most  Lysis of RBC = laking/ hemolysis (in vivo or in vitro)
 easy to manipulate to clot chemistry,  In vitro hemolysis is due to (common):
 ideal for peripheral blood smears immunologic 1. use of vacuum tubes
 less intimidating & blood bank 2. vigorous mixing
Disadvantages of skin puncture: tests 3. use of too narrow/ roo wide needle bores
 less amt of blood obtained Red/ None; Allows blood Most 4. effect of ROH
 add & repeated tests can’t be done gray or separator to clot; chemistry 5. centrifugation & separation steps
 blood hemolyzes easily red material barrier tests  Hemolysis is visible only not until a 200mg/L of Hb level
 painful (finger) black between  Ictersia (Icteric serum)
Procedure: cells &  Intensely yellow serum sample (elevated Bb value)
1. ID patient serum  Jaundice: Bb greater than 430uM (25mg/L)
2. reassure Orange thrombin Accelerated STAT serum  Bb interferes w/ test using dyes & turbidity test
3. assemble equipment clot test  Interference due to Bb may be minimized: sample
4. prepare finger Blue Buffered Binds Ca Coagulation blanking or dual wavelength method (Allen correction
5. puncture finger citrate assays like PT method)
6. eliminate 1st drop & APTT  Lactescene (Lipemic serum)
7. produce large rounded drop Black Buffered Binds Ca Westergren  Obtained normally after a meal (elevated exogenous
8. withdraw/collect blood sodium ESR chylomicrons)
9. stop bleeding citrate  Characterized by milky or highly turbid serum
10. thank patient Gray  NaF/  Inhibits Glucose  Lactescene appears when TAG level reaches 4.6mm (4g/L)
11. transport specimen to lab K2C2O4 glycolytic determination  Corrected by ultracentrifugation of serum sample
Things to remember in doing skin puncture: enzymes  Grounds for rejecting specimen
 depth: 2.5-3mm  Iodoacetat enolase &  Inadequate sample ID
  Insufficient volume of specimen collection
 Inappropriate collection tube
 Hemolysis
 Improper transportation
 Interferences
Things to remember:
 All materials: dry & sterile (avoid hemolysis)
 Never puncture/ draw blood from vein where IV medication
is running (false low)
 Site of puncture: thoroughly clean (avoid Thrombophlebitis)
 Tourniquet: not tied too tight (constrict arteries & veins)
 Tourniquet: always released before withdrawing needle
from vein
 Remove needle from adaptor of syringe & allow blood to flow
gently down sides of tube
 Container: stoppered; AC tubes inverted 6-10x
 Reusable syringes: rinsed w/ tap H2O
 Needles: sharp container