PRINCIPLES OF MEDICAL LABORATORY SCIENCE 2

College of Allied Health Sciences

Bachelor of Science in Medical Laboratory Science
Second Semester, A.Y. 2022-2023

[TRANS] PMLS UNIT 9: SPECIMEN HANDLING AND PROCESSING

o Failure to follow temperature requirements
OUTLINE o Transport method (Hand/Pneumatic Tube)
 During Specimen Processing
I Specimen Handling o Contamination
II Routine Handling o Delay in processing or testing
III Special Handling o Delay in fluid separation from cells
IV Special Processing
o Evaporation
V Specimens Not Requiring Further Processing
VI Specimens That Require Centrifugation o Failure to centrifuge specimen according to test
requirements
o Failure to separate fluid from cells
o Incomplete centrifugation
SPECIMEN HANDLING
o Mislabeled aliquot
 It has been estimated that 46% to 68% (32%-75%, o Aliquot - a portion or fraction of a biological sample
McPherson et. al., 2011) of all laboratory errors occur that is taken or divided for analysis, testing, or
prior to analysis. storage purposes.
 3 phases of the testing process: o Multiple centrifugations
o Preexamination / Preanalytical o Rimming of clots
o Examination / Analytical  During Specimen Storage
o Postexamination / Postanalytical o Exposure to light
 Proper handling from the time a specimen is collected until o Temperature change outside defined limits
the test is performed helps ensure that results obtained on
the specimen accurately reflect the status of the patient.
ROUTINE HANDLING
POSSIBLE SOURCES OF MIXING TUBES BY INVERSION
PREEXAMINATION/PREANALYTICAL ERROR  Gentle inversion helps to distribute the additive evenly
 Before Collection while minimizing the chance of hemolysis.
o Altitude  Examples of tests that cannot be performed on hemolyzed
o Dehydrated Patient specimens
o Duplicate test orders o Potassium, magnesium, and most enzyme tests
o Exercise
 Inadequate mixing of anticoagulant tubes leads to
o Inadequate Fast
microclot formation.
o Incomplete Requisition
 Inadequate mixing of gel separator tubes may prevent the
o Medications
additive from functioning properly, and clotting may be
o Patient Stress
incomplete.
o Pregnancy
o Smoking  Nonadditive tubes do not require mixing.
o Strenuous Exercise
o Treatments TRANSPORTING SPECIMENS
o Wrong test ordered  Tubes should be transported stopper up to reduce
 At Time of Collection agitation, aid clot formation in serum tubes, and prevent
o Misidentified Patient contact of the tube contents with the tube stopper.
o Antiseptic not dry  CLSI and OSHA guidelines require specimen transport
o Expired Tube bags to have a biohazard logo, a liquid-tight closure, and
o Failure to invert additive tubes properly a slip pocket for paperwork.
o Faulty technique  Nonblood specimens should be transported in leak-proof
o Improper vein selection containers with adequately secured lids.
o Inadequate volume of blood  All specimens transported through pneumatic tube
o Inappropriate use of plasma separator tube (PST) or systems should be protected from shock and sealed in
Serum Separator Tube (SST) zipper-type plastic bags to contain spills.
o Incorrect Collection Tube o Pneumatic Tube System
o Incorrect needle position  Specimens sent via pneumatic tube system
o Incorrect needle size must be in zippertype plastic bags to contain
o Mislabeled Tube spills and protected from shock.
o Mixing tubes too vigorously  Robot specimen transportation system called the
o Nonsterile site preparation RoboCourier® Autonomous Mobile Robot
o Patient position
o Prolonged tourniquet application DELIVERY TIME LIMITS
o Underfilled tube  Stat specimens must be transported, processed, and
o Wrong collection time tested immediately.
 During Specimen Transport  Routine blood specimens should ideally be delivered to
o Agitation-induced hemolysis the laboratory within 45 minutes of collection and
o Delay in transporting centrifuged within 1 hour of arrival if serum or plasma is
o Exposure to light needed.

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 The maximum time limit for serum or plasma separation
according to CLSI standards is 2 hours after collection.
 Nonadditive and gel-barrier serum tubes, such as SSTs,
must be completely clotted before centrifugation.
 Heparin gel-barrier tubes, such as PSTs, can be
centrifuged right away.
 Specimens in gel barrier tubes do not require manual
separation after they have been centrifuged (separator gel
prevents glycolysis for up to 24 hours).
TIME LIMIT EXCEPTIONS
 “STAT” or “medical emergency” specimens take priority
over all other specimens.
 Examples of other exceptions to time limits:
o Ammonia specimens - immediately placed in ice
slurry or a cooling tray after collection; transport
STAT; separate from the cells within 15 minutes of
collection
o Coagulation specimens – perform PT and APTT
within 4 hours of specimen collection; frozen at -
20°C or below for up to two weeks or –70°C for
longer storage
o Glucose specimens in Sodium Fluoride tubes -
stable for 24 hours at room temperature and for up
to 48 hours when refrigerated at 4° to 8°C
o PT results on unrefrigerated and uncentrifuged
specimens are reliable for up to 24 hours after
collection.
o APTT test specimens require analysis within 4
hours of collection regardless of storage conditions.
o Pediatric glucose specimens – test ASAP; difficult
to inhibit glycolysis
 Hematology specimens
o Blood smears (from EDTA specimens) – prepare
within 1 hour of collection
o EDTA specimens for CBCs - analyze within 6 hours;
stable for 24 hours at RT
o EDTA specimens for ESR determinations – test
within 4 hours if left RT or within 12 hours if
refrigerated.
o EDTA specimens for reticulocyte count – stable up
to 6 hours at RT and up to 72 hours if refrigerated
 Molecular test specimens – RNA substances are
unstable; batch analysis - store plasma at 4°C for 48 hours;
if not analyzed within 48 hours, transfer into an aliquot tube
and store at -80°C
 Microbiology specimens
 Urine specimens – can be held at RT for up to 2 hours
when not transported or analyzed promptly
SPECIAL HANDLING
TEMPERATURES RELATED TO SPECIMEN
HANDLING:
 Body temperature: 36.4°C–37.6° C (37°C average)
 Room temperature: 20°C–30°C (25°C average)
 Refrigerated temperature: 2°C–10°C
 Frozen temperature: −20°C or lower (some specimens
require−70 °C or lower)
BODY TEMPERATURE SPECIMENS
 Specimens need to be transported at or near normal body
temperature of 37ᵒC
 Collect in a tube that has been prewarmed to 37ᵒC
 Small, portable heat blocks kept in 37ᵒC incubator
 Can hold this temperature for about 15 minutes after
removal from the incubator
 Specimens can be wrapped in an activated heel warmer
for transport
CHILLED SPECIMENS
 Blood specimens that require chilling should be completely
immersed in a slurry of crushed ice and water.
 Large cubes or chunks of ice without water added do not
allow adequate cooling of the entire specimen.
SPECIMENS THAT MUST NOT BE CHILLED
 Coagulation specimens - can activate clotting factors
and disrupt platelet function
 Potassium - cold inhibits glycolysis
 Arterial blood gas - should not be placed on ice if analyzed
within 30 minutes of collection
LIGHT-SENSITIVE SPECIMENS
 Bilirubin can decrease up to 50% after 1 hour of light
exposure
 Wrap specimen in aluminum foil
 Amber-colored microcollection containers – infant
bilirubin specimens
 Amber containers – urine collection
SPECIMEN SUITABILITY
 The most frequently cited reason for rejection of chemistry
specimens is hemolysis, followed by insufficient amount of
specimen or QNS (quantity not sufficient).
 The most frequent reason for rejection of hematology
specimens is clotting.
EXAMPLES OF SPECIMEN REJECTION CRITERIA
 Inadequate, inaccurate, or missing specimen identification
 Additive tubes containing an inadequate volume of blood
 Hemolysis
 Wrong Tube
 Outdate Tube
 Improper Handling
 Contaminated Specimen
 Insufficient specimen, reffered to as "quantity not sufficient"
(QNS) for the test ordered
 Wrong collection time
 Exposure to light
 Delay in testing
 Delay or error in processing. Serum tubes that have not
been spun within 2 hours or refrigeration of serum tubes
before centrifugation will increase some analytes, such as
potassium, creatinine, phosphorus, LDH, and decrease
analytes, such as glucose, ionized calcium, and CO₂
SPECIMENS NOT REQUIRING FURTHER
PROCESSING (E.G. CENTRIFUGATION,
SEPARATION)
 CBCs and other hematology specimens drawn in EDTA
tubes
 Other whole-blood specimens for tests such as
cyclosporine, hemoglobin A1c, and whole-blood lead
analysis
 Cerebrospinal fluid (CSF), serous fluids, synovial fluid
and bronchoalveolar lavage (BAL) specimens
 Microbiology specimens
 Urinalysis specimens
SPECIMENS THAT REQUIRE CENTRIFUGATION
 3 Phases:
o Precentrifugation
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 After specimen collection and before  Glucagon Urine
centrifugation  Homocysteine porphobilinogen
o Centrifugation  Lactic acid
 When the specimen is in the centrifuge  Parathyroid
o Postcentrifugation Hormone (PTH)
 After centrifugation and before removal of  pH/blood gas
serum or plasma  Pyruvate
 Renin
SPECIMEN PROCESSING

CENTRIFUGATION
 A centrifuge is a machine that spins blood and other
specimens at a high number of revolutions per minute
(rpm).
 It is crucial that tubes be “balanced” in a centrifuge.
 Place equal-size tubes with equal volumes of specimen
opposite one another in the centrifuge.
 A specimen should never be centrifuged more than once.
 Centrifuging plasma specimens:
o May be centrifuged without delay
o For STAT chemistry tests
 Centrifuging serum specimens:
o Must be completely clotted (30-60 mins at RT)
before centrifugation
o SSTs and tubes containing clot activators clot
within 30 mins
o Thrombin tubes (like BD rapid serum tube) clot in 5
mins
 NOTE: Stoppers should also be left on tubes during
centrifugation to prevent contamination, evaporation,
aerosol (fine spray) formation, and pH changes.

ALIQUOT PREPARATION
 An aliquot is a portion of a specimen used for testing.
 Chemistry - BUN, Creatinine, AST, ALT, GGT, ALBUMIN
 Serology – HBsAg , Anti-HAV, Anti-HCV
 Aliquots are prepared by transferring a portion of the
specimen into one or more tubes labeled with the same
ID information as the specimen tube.
 NOTE: According to OSHA, “All procedures involving
blood or potentially infectious materials shall be
performed in such a manner as to minimize splashing,
spraying, splattering, and generation of droplets of
these substances.”

 Caution: Never put serum and plasma, or plasma from

specimens with different anticoagulants, in the same
aliquot tube.
 Caution: Pouring the serum or plasma into aliquot tubes is
not recommended because it increases the possibility of
aerosol formation of splashing

EXAMPLES OF SPECIMENS THAT REQUIRE

SPECIAL HANDLING

Keep at 37ᵒC Chill in Crushed Protect from

Ice Slurry Light
Cold Agglutinin Adrenocorticotropic Bilirubin
Hormone (ACTH)
Cryofibrinogen Acetone Carotene
Cryoglobulins  Angiotensin- Red cell folate
converting Serum folate
enzyme(ACE) Vitamin B₂
 Ammonia Vitamin B₆
 Catecholamines Vitamin B₁₂
 Free fatty acids Vitamin C
 Gastrin Urine porhphyrins

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