BLOOD COLLECTION METHODS AND PROCEDURE FOR STUDENTS

BLOOD COLLECTION METHODS AND PROCEDURE

1. BLOOD COLLECTION
NEETHU SOMAN
MSc Medical Biochemistry

2. COLLECTION OF BLOOD
•Blood collection is three types.
1.Arterial blood collection : Carrying oxygenated blood from heart to several
parts of the body by the pumping action Of heart.
2.Venous blood collection: Carrying deoxygenated blood from distant parts of
the body to heart & to lungs.
3.Capillary blood collection:a minute blood vessel that connects the smallest
arteries to the smallestveins and serves as an oxygen

3. CAPILLARY BLOOD COLLECTION
•Capillary puncture, also called dermal puncture, is a safe, rapid, and efficient
means of collecting a blood specimen.
•To perform capillary puncture, a small sterile lancet or blade is used to puncture
the skin and capillaries to create a blood flow.
•Capillarypunctures are performed when only a small amount of blood is required,
when obtaining blood from infants, or when the patient has a condition that makes
venipuncture difficult.
•In the clinical laboratory, capillary blood has been used only in special situations,
because of the small sample volume and potential for rapid clotting of the sample.
•However, the increased use of small, portable, easy-to-use instruments that
require only a drop or two of blood, has made capillary blood the specimen of
choice for these analyzers.

4. Capillary Puncture Sites
•Theusualsiteforcapillary
punctureinadultsandchildrenis
thefingertip.
•Inadults,theringfingerisoften
selectedbecauseitusuallyisless
calloused.
•Fornewbornsandinfants,capillary
bloodcanbeobtainedfromthe
lateralorsideportionoftheheel
pad.Onceaninfantbeginstowalk
(abouttheageof1year)blood
shouldbecollectedfroma
fingerstick.

5. Capillary Puncture Equipment
Lancets:Severaldesignsofdisposable,
sterilesafetylancetsareavailablefor
capillarypuncture.
•Theselancetsmakepuncturesof
uniformdepthatthetouchofabutton
andareavailableinseveralblade
lengthsforuseindifferentsituations.
•Specialpediatriclancetsthatproducea
shallowpunctureshouldbeusedwith
infants.

6. Capillary Blood Collection Equipment
•The following are needed for routine capillary blood collection:
1.Sterile lancet—Retractable type is preferred; tip less than 2.4 mm. The tip length will depend
on the intended use. For newborns and infants under 12 months, the device should cut or
puncture no more than 0.85 mm. The lancet be inserted at during capillary blood collection at
45 degrees .
2.Alcohol wipes—Wipes containing 70% isopropyl alcohol.
3.Sterile gauze pads—For removal of first free-flowing drop of blood and for pressure
application after collection.
4.Gloves—Can be made of latex, rubber, vinyl, etc.; worn to protect the patient and the
phlebotomist.
5.Eye protection—To protect the phlebotomist from aerosols contaminating the eyes.
6.Sharps disposal unit—Lancets must be placed in a proper disposal unit immediately after
their use.

7. •CapillaryCollectionContainers:Capillary
bloodcanbecollectedincapillarytubesor
collectionvials.Capillarybloodrequiredfor
testsotherthanmicrohematocrit,suchas
chemistrytests,canbecollectedinspecial
vialswithacapillaryorotherextensionfor
directingthebloodintothevial.
•Thesevialsareavailableplainorwith
anticoagulant.Usingthesevials,asmall
quantityofwholeblood,plasma,orserum
canbeobtained

8. Collecting the Blood Sample
•Thefirstdropofbloodshouldbewipedawaywithdry,sterilegauze.
•Thisfirstdropcontainstissuefluid,whichdilutestheblooddropandcanalsoactivate
clotting.
•Thesecondandfollowingdropsofbloodareusedforthetestsample.
•Awell-roundeddropofbloodshouldbeallowedtoformbeforecollectionbegins.
•Thehandcanbegentlymassagedtoincreasebloodflow,butexcessivepressurenearthe
puncturesiteshouldbeavoided.(Squeezingthefingertipcanforcetissuefluidintotheblood
sample.)
•Capillarybloodshouldbecollectedasquicklyaspossibletopreventclotting.
•Thecapillarytubeshouldbeheldinanalmosthorizontalposition,ortiltedslightlydownward;
thebloodcollectingvialshouldbeheldverticallysobloodwillflowdownintothetube.
•Whenthetipofthecapillarytubeistouchedtotheblooddrop,bloodwillenterthetubeby
capillaryactionbecauseoftheattractionbetweentheliquidandthetube.
•Capillarytubesshouldbefilledthree-quartersfull

9. PERFORMING THE CAPILLARY PUNCTURE
(A)performpunctureon
cleansedfingertip;wipeaway
firstblooddropand
(B)allowroundeddropof
bloodtoform;
(C)collectbloodinto
capillarytubeor
(D)collectionvial

10. Caring for the Capillary Puncture Site
•After the blood has been collected, sterile gauze or a cotton ball should
be placed on the puncture site and pressure applied until bleeding
stops.
•A small adhesive bandage can be applied if necessary.

11. PRECAUTIONS
1.Alwayswearlabcoatandgloveswhilecollectingbloodsample.
2.Donotsqueezepuncturesitetootightlyasthiscandilutethesample.
3.Avoidtouchingbloodspecimenandpuncturesite.
4.Puncturedepthshouldbelessthan1.5mm.
5.Usemiddleorringfingerofhandforcapillarybloodcollection.
6.Nevertouchtipofthelancet.
7.Alwaysusedisposableglovesanddisposablelancet.

12. VENOUS BLOOD COLLECTION
•Venipunctureisacommonmethodofobtainingbloodforlaboratory
examination.
•Thevenipunctureisaquickwaytoobtainalargesampleofbloodonwhichmany
differentanalysescanbeperformed.
•Inavenipuncture,alsocalledaphlebotomy,asuperficialveinispuncturedwith
ahypodermicneedleandbloodiscollectedintoavacuumtubeorsyringe.
•Thevenipunctureisasafeprocedurewhenperformedcorrectlybytrained
personnel.
•Thesepersonnelcanincludephlebotomists,medicalassistants,nursingstaff,
physicians,medicallaboratorytechnicians,andmedicallaboratoryscientists.
•Thevenipuncturemustbeperformedcarefullytopreservetheconditionofthe
vein.Muchobservationandpracticeunderthesupervisionofanexperienced
phlebotomistisrequiredtobecomeskilledandself-confidentintheartof
venipuncture

13. •Performingavenipunctureinvolvesseveralimportantstepsthatmustbe
thoroughlyunderstoodbeforetheprocedureisattempted:ObservingStandard
Precautionsandothersafetymeasuresthroughoutprocedure.
•Selectingtheproperequipment
•Identifyingthepatientusingtwoidentifiers
•Preparingthepatientforvenipuncture
•Selectingandpreparingthepuncturesite
•Applyingandremovingthetourniquet
•Obtainingtheblood
•Caringforthepuncturesite
•Observingthepatientforadversereaction
•Labelingbloodspecimensimmediatelyfollowingbloodcollection

14. VENIPUNCTURE MATERIALS AND SUPPLIES
•Venipuncture can be performed using a safety needle/collection tube holder
assembly , a safety needle and syringe, or a winged collection set with tubing
and tube holder .
•Other materials required for venipuncture include evacuated blood collecting
tubes, alcohol swabs, sterile gauze,disposable tourniquet, and small adhesive
bandage.
•Venipuncture is performed using a vacuum-tube system, a syringe, or a winged
collection set. The safety guidelines and quality assessment considerations are
the same for all venipuncturemethods.

15. VENIPUNCTURE MATERIALS AND SUPPLIES
1.SafetyNeedles,22gorless
2.Butterflyneedles.21gorless
3.Syringes
4.Vacutainertubeholder
5.TransferDevice
6.BloodCollectionTubes.
•Thevacuumtubesaredesignedtodraw
apredeterminedvolumeofblood.
•Tubeswithdifferentadditivesareused
forcollectingbloodspecimensfor
specifictypesoftests.
•Thecolorofcapisusedtoidentifythese
additives.
7.Tourniquets.Singleuse,disposable,
latex-freetourniquets
8.Antiseptic.Individuallypackaged
70%isopropylalcoholwipes.
9.22Gauze
10.SharpsDisposalContainer.An
OSHAacceptable,punctureproof
containermarked“Biohazardous”.
11.Bandagesortap

16. VENIPUNCTURE MATERIALS AND SUPPLIES

17. PROCEDURE
1.Identifythepatient,twoformsofactiveidentificationarerequired.
•Askthepatienttostatetheirnameanddateofbirth.
•Thisinformationmustmatchtherequisition.
2.Reassurethepatientthattheminimumamountofbloodrequiredfortestingwill
bedrawn.
3.Verifythatanydietortimerestrictionshavebeenmet.
4.OrderofDraw
•ThefollowingorderofdrawistheapprovedorderasestablishedbyCLSI.
•Thisorderofdrawshouldbefollowedwhenevermultipletubesaredrawnduring
asinglevenipuncture.
•Thisistopreventcrosscontaminationbythetubeadditivesthatcouldleadto
erroneousresults

18. 1. Assemble the necessary equipment appropriate to the
patient’s physical characteristics.
2. Wash hands and put on gloves.
3. Position the patient with the arm extended to form a
straight-line form shoulder to wrist.
4. Do not attempt a venipuncture more than twice. Notify
your supervisor or patient’s physician if unsuccessful.

19. 5.Selecttheappropriateveinforvenipuncture.
•Thelargermediancubital,basilicandcephalicveinsaremostfrequentlyused,but
othermaybenecessaryandwillbecomemoreprominentifthepatientcloseshisfist
tightly.
•Atnotimemayphlebotomistsperformvenipunctureonanartery.
•Itisnotrecommendedthatbloodbedrawnfromthefeet.TheProviderspermissionis
requiredtodrawfromthissite.
•Extensivescarringorhealedburnareasshouldbeavoided
•Specimensshouldnotbeobtainedfromthearmonthesamesideasamastectomy.
•Avoidareasofhematoma.
•IfanIVisinplace,samplesmaybeobtainedbelowbutNEVERabovetheIVsite.
•Donotobtainspecimensfromanarmhavingacannula,fistula,orvasculargraft.
•Allow10-15minutesafteratransfusioniscompletedbeforeobtainingabloodsample

21. 6. Apply the tourniquet 3-4 inches above the collection site.
•Never leave the tourniquet on for over 1 minute.
•If a tourniquet is used for preliminary vein selection, release it and reapply after
two minutes.
7. Clean the puncture site by making a smooth circular pass over the site with the
70% alcohol pad, moving in an outward spiral from the zone of penetration.
•Allow the skin to dry before proceeding.
•Do not touch the puncture site after cleaning.

22. 8. Perform the venipuncture
A. Attach the appropriate needle to the hub by removing the plastic cap over the small end of the needle and
inserting into the hub, twisting it tight.
B. Remove plastic cap over needle and hold bevel up.
C. Pull the skin tight with your thumb or index finger just below the puncture site.
D. Holding the needle in line with the vein, use a quick, small thrust to penetrate the skin and enter the vein in one
smooth motion.
E. Holding the hub securely, insert the first vacutainer tube following proper order of draw into the large end of the
hub penetrating the stopper. Blood should flow into the evacuated tube.
F. After blood starts to flow, release the tourniquet and ask the patient to open his or her hand.
G. When blood flow stops, remove the tube by holding the hub securely and pulling the tube off the needle.
H. Gently invert each tube
Light blue top-invert 3-4 times
Red and gold tops invert 5 times.
All other tubes containing an additive should be gently inverted 8-10 times.

23. 9. Place a gauze pad over the puncture site and remove the needle.
10. Activate the safety device and properly dispose of the vacutainer holder with needle attached
into a sharps container.
11. Immediately apply slight pressure to the gauze pad over the venipuncture site..
•Ask the patient to apply pressure for at least 2 minutes.
•When bleeding stops, apply a fresh bandage, gauze or tape.
12. Tubes must be positively identified after filling with a firmly attached patient label.
•The label must include the patient first and last names, DOB,, collection date and time and
collectors initials.
•If no patient labels are available, manually label the tubes with the required information. All
labels must include two identifiers .
•The tube must be labeled before leaving the patient.
13. Observe special handling requirements
•Some test specimens require special handling for accurate results.

24. Anticoagulants and Additives
•Thestandardoperatingprocedure(SOP)manualwillincludealistoftestsperformedandthe
typeandsizeofvacuumtubethatshouldbeusedwhencollectingbloodforeachtest.Thecap
colorsofthebloodcollectiontubesdesignatewhich,ifany,anticoagulantoradditiveispresent
.Whenmultipletubesmustbefilledduringavenipuncture,aparticularorderofdraw,suchas
shown,mustbeused.
•Fillingtubesinthecorrectorderpreventscrosscontaminationofanticoagulantsbetween
tubes,whichcouldadverselyaffecttestresults.Whenthetubestopperorcapispiercedbythe
bluntendofthevenipunctureneedle,thevacuumdrawsthebloodintothetube.Itisimportant
thattubesbefilledtotheirstatedcapacitiesbecauseanincorrectblood:anticoagulantratiocan
altercellmorphologyandcauseerroneoustestresults,especiallyincoagulationtests.
•Sometubeshaveafilllinemarkedonthetubetoeasilydetermineifbloodvolumeis
sufficient.Forcertaintests,tubeswithanticoagulantcanstillbeusedfortestsiftheyarenot
completelyfilledbutareatleast70%full.
•Thelaboratory’sSOPmanualwillspecifythetestsforwhichtheleveloftubefilliscritical.
Forexample,onerequirementforcoagulationspecimenacceptabilityisthatcitratetubesmust
befilledtoatleast90%capacity.

26. When collecting multiple specimens, blood tubes should be drawn in the
following order; mix all tubes by inversion 6 –8 times:
1. Blood Culture bottles
2. Isolator tube
3. Blue top (3.2% sodium citrate)
4. Red top (no preservative)and Gold top (SST)
5. Royal blue top (no preservative)
6. Green top (sodium heparin)
7. Lavender top (EDTA), Pink top (EDTA), and Royal
blue top (EDTA)
8. Gray top (sodium fluoride)
9. Yellow top (ACD) Solution A or B
10. TB Gold QuantiFERON: Nil (gray top), TB antigen
(red top), and Mitogen (purple top)

27. Arterial blood Collection
•ArterialbloodisthepreferredspecimenforBloodGasdeterminations.
•Arterialpuncturesaretechnicallydifficultwithpotentiallymoreriskforthe
patientthanvenipuncture;therefore,inclinicalpractice,itisreservedforthe
mostessentialpurposes.
•Conditionsthatcommonlyutilizethemeasurementofbloodgasesinclude
chronicobstructivepulmonarydisease(COPD),cardiacandrespiratoryfailures,
severeshock,lungcancer,diabeticcoma,coronarybypass,openhearsurgery,
andrespiratorydistresssyndrome(RDS)inprematureinfants.
•Patientsrequiringbloodgasdeterminationsareoftencriticallyillandrequire
vigorousmeasurestosupportlifethroughassistedventilationusingmixturesof
gasestailored,inresponsetolaboratoryfindings,toserveindividualneeds.

28. SPECIMEN REQUIREMENTS:
•A standard blood gas collection kit is used in all areas. The collection
kit utilizes dry lithium heparin and a syringe volume preset collection
system.
•Routine practice is to preset the volume at 3 mL.for arterial blood gas
collections. and to expel any air in the syringe after sample collection
and to transport on ice to the central laboratory.

29. PROCEDURE:
Patient Preparation
A. The phlebotomist must wash their hands, prior to attending to the patient, with
either soap and water or “waterless soap.”
B. Patient identification is made by cross-checking patient orders against patient
arm band and room number for accuracy.
C. Patient is informed that the collection is an arterial puncture requested by their
physician. Inform them that the puncture could be more painful than a
venipuncture and that the site must be held with pressure for a at least 5 minutes.
If the patient is on anticoagulants, it may require additional holding time until
bleeding has completely stopped. If bleeding persist longer than 10 minutes
contact the patients nurse.
D. Obtain and record preliminary patient information

30. Site Selection
•A.Radialartery.Theradialarteryisthepreferredsiteforarterialpunctureasthe
ulnararteryprovidescollateralcirculationtothewristanditisclosetothesurface
ofthewristandeasilyaccessible.Ifbothradialarteriesareunacceptableforuse
thenextsitewouldbethebrachialartery.
•B.Brachialartery.Thissiteisreservedforsecondaryuseasthemedianand
ulnarnervesandthebasilicveinareincloseproximitytothebrachialartery.The
femoralisthelastsiteofchoice
•C.Femoralartery.Infectionisariskatthissiteanditmaybemoredifficultto
controlthebleedingatthissitepostpuncture.Pressuremustbeappliedatthissite
foraminimumof10minutes.Femoralpuncturesareonlydonebyrequestofthe
physician.Theyareperformedmostfrequentlyduring“Code99”becauseitis
usuallytheeasiestsitetopalpateandpuncture.

31. Performing the Puncture
A. Prepare the syringe by setting the preset volume and positioning the needle with
the bevel side up.
B. Enter the artery at a 45 degree angle (radial) (90degree for brachial) slowly, until
blood appears in the needle hub. If blood does not appear, the needle may be
slightly redirected but remain under the skin. NOTE: If you are unsuccessful after
two attempts then withdraw the needle and allow the patient to rest. Allow someone
else to perform the puncture. Use a new syringe and needle with each attempt.
C. When the preset volume has been collected, remove the needle and apply firm
pressure for a minimum of 5 minutes (5 for radial or brachial, 10 for femoral
puncture) by placing the sterile gauze pad over the puncture.
D. Expel any air from the syringe, replace the needle with the syringe cap provided,
and rotate the syringe to insure thorough mixing of the sample with the dried
heparin.

32. LIMITATIONS OF PROCEDURE:
•SOURCESOFSAMPLEERROR:
A.Airinthesamplesyringe--Theatmosphericoxygenentersthespecimenand
carbondioxidefromthespecimenenterstheairbubble.
B.ClottedSample--Causedbyinadequatemixingofthespecimenwith
anticoagulantaftercollection.
C.Delayintestingasamplenotonice--Bloodcellmetabolismcontinues
utilizingoxygenandproducescarbondioxideandacidsthatlowerpH.
D.Insufficientpatientinformationdocumentation--Nopatienttemperature.No
oxygentherapyinformation.SiteofpunctureandAllentestnotrecorded.
E.Patienthyperventilationduringprocedurecancausefalsechangesin
metabolicstatus.

33. REFFERENCE
•Basic clinical laboratory techniques-Barbara h. Estridge
•Concise book of medical laboratory technology methods and
interpretation –RamanikSood
•Text book of medical laboratory technology –Godkar
•Haematology -RamanikSood