CYPRUS SCIENCE UNIVERSITY

FACULTY OF HEALTH SCIENCE

BIOCHEMISTRY
BIO – 102

BIOLOGICAL
LABORATORIES
A S S I S T. P R O F. D R . C A N G Ü L T U N C AY A R N AV U T
Introduction to clinical laboratories:
Clinical labs are important in diseases diagnosis, determination its
severity and patient response to specific treatment.
Diagnosis of any disease is first done by physical examination by
clinician and confirmed by lab diagnostic tests.
Lab values are very important in determination of disease severity,
drug doses and in follow up.
Introduction
The sections of clinical laboratory are:
Clinical pathology
Hematology
Clinical biochemistry
Clinical microbiology
Serology
Blood bank
Histology and cytology
Introduction
Clinical biochemistry:
It deals with the applications of biochemistry laboratory to
find out the cause of a disease.

Types of samples that are used in testing:

Body fluids: blood, serum, plasma, urine, cerebrospinal fluid
(CSF), feces, and other body fluids or tissues.
Samples
Blood samples and specimens
Most clinical laboratory assays are designed to detect or
quantify substances or cells which are called analytes
Blood must be collected and proccessed properly to avoid
artifactual changes
Blood withdrawn from a vessel must be immediately mixed
with an anticoagulant to prevent clot formation and/or keep
the cells and other components in suspension
Plasma is the fluid component of the blood that is collected
after the centrifugation
Samples
Blood samples and specimens
- Anticoagulants
- Calcium binding agents (EDTA, Citrate, Oxalates)
- Heparin
- Serum is the fluid component of blood that is collected after coagulated blood
sample

- Urine Samples
- Other Body fluids
Biochemical tests in clinical medicine
Lipid profile
Diabetic profile
Kidney profile
Liver profile
Bone profile
Electrolyte profile
 Lab request and lab report forms
Lab request form: it fills computerize or paper filled by the doctor then send it to the
lab. The lab request contains a list of tests to be performed on specimen of patient. Each
lab has its specific request; for example, chemistry request, hematology request… etc.
Lab report form: it contains the result of patient.
Laboratory work flow cycle:
The flow cycle includes the entire steps of laboratory test, starting from test ordering by
a doctor until reporting the results.
Three phases of laboratory testing:
Pre-analytical: test ordering, specimen collection, transport and processing
Analytical-testing
Post-analytical: testing results transmission, interpretation, follow-up, retesting.
 Disposable syringes Vacationer systems Disposable lancets

Phlebotomy
Phlebotomy or blood collection:
The term phlebotomy refers to blood
draw from a vein, artery, or the Gauze pads absorbent cotton Tourniquet
capillary bed for lab analysis or blood
transfusion.

The phlebotomy equipments: Alcohol swap Plastic bandage Waste container

For specimen collection, the following

materials will be required:
Preparation of Blood Sample
One of three different specimens may be used:
whole blood
serum
plasma
First: Whole-blood specimen:
It must be analyzed within limited time (why?)
Over time, cells will lyse in whole-blood which will change the
conc. of some analytes as potassium, phosphate and lactate
dehydrogenase.
Some cellular metabolic processes will continue which will
alter analytes conc. like glucose and lactate.
Serum
2. Serum:
Difference between Serum and plasma:
Serum is the same as plasma except it doesn't contain
clotting factors (as fibrin).
Plasma contains all clotting factors.
So, serum and plasma all has the same contents of
electrolytes, enzymes proteins, hormones except clotting
factors
Serum is mainly use in chemistry lab & serology.
Plasma
3. Plasma:
The tube will have anti-coagulation
After centrifugation the blood sample got
separated into three layers
 In the lab
Specimen rejection criteria:
Specimen improperly labeled or unlabeled
Specimen improperly collected or preserved
Specimen submitted without properly completed
request form
Hemolyzed sample (show tubes)
 Hemolysis
It means releaseof hemoglobin due to rupture of RBCs.
Due to hemolysis plasma or serum appears pink to red color.
It causes erroneously high: K+, Ca2+, phosphate, AST and SLDH.
Hemolysis is occurred due to sampling, transporting and storage (too hot or too
cold).
According to the degree of hemolysis it is classified as H+, H++ and H+++. H+ may be
accepted for some tests that are not affected by RBCs contents as glucose and lactate,
H++ and H+++ not acceptable for any test.

Changes in the serum color indicate one of the following:

Hemolyzed: serum appears pink to red due to rupture of RBCs
Icteric: serum appears yellow due to high bilirubin.
Lipemic: serum appears milky or turbid due to high lipid.
 Collection Tubes
The most widely used tubes for blood collection are evacuated
tubes (Vacutainers)

◦ Negative pressure facilitates collection

◦ Easy to use
◦ Sterile
◦ Universally used colour-coded rubber stoppers to denote tube type.
◦ Tubes can contain various anticoagulants for the collection of whole blood
or plasma.
◦ Tubes can have additives for specific tests (glucose, metals)
 Collection Tubes
(Vacutainers)

Separator Gel

Serum
Separator Gel

Serum Separator Tube (SST) Clot

Blood collection tubes:

Two major types of blood collecting tubes:

Serum separating tubes (SST)
Plasma separating tubes (PST)
Collection tubes

Red-top tubes contain no anticoagulants or preservatives

Red-top tubes are used for collecting serum
◦ 10-15 minutes is required to allow blood to clot before centrifuging
◦ Used for blood bank specimens, some chemistries
Collection tubes

Gold (and “tiger”) top tubes contain a gel that forms a physical barrier
between the serum and cells after centrifugation
No other additives are present
Gel barrier may affect some lab tests
Collection tubes

Used for Glucose measurement.

After blood collection, glucose concentration decreases
significantly because of cellular metabolism
Gray-top tubes contain either:
◦ Sodium fluoride and potassium oxalate, or
◦ Sodium iodoacetate
Both preservatives stabilize glucose in plasma by inhibiting
enzymes of the glycolytic pathway
◦ NaF/oxalate inhibits enolase
◦ Iodoacetate inhibits glucose-3-phosphate dehydrogenase
Collection tubes

Green-top tubes contain either the Na, K, or lithium (Li) salt

of heparin. Most widely used anticoagulant for chemistry
tests.
◦ Should not be used for Na, K or Li measurement
◦ Can effect the size and integrity of cellular blood components and not
recommended for hematology studies
Heparin accelerates the action of antithrombin III, which
inhibits thrombin, so blood does not clot (plasma)
The advantage of plasma is that no time is wasted waiting
for the specimen to clot
Collection tubes

Lavender-top tubes contain the K salt of ethylenediaminetetraacetic acid

(EDTA), which chelates calcium (essential for clot formation) and inhibits
coagulation
Used for hematology, and some chemistries
Cannot be used for K or Ca tests
Collection tubes

Blue-top tubes contain sodium citrate, which chelates calcium and

inhibits coagulation
Used for coagulation studies because it is easily reversible.
Collection tubes

Brown and Royal Blue top tubes are specially cleaned for trace metal
studies
◦ Brown-top tubes are used for lead (Pb) analysis
◦ Royal blue-top tubes are used for other trace element studies (acid washed)
Blood collection tubes:
 Plasma Separating Tubes (PST)
Top Color Additives Principle Uses
Lavender EDTA -The strongest anti-coagulant - Hematology
- Ca+2 chelating agent - Blood bank (ABO)
- To preserve blood cells components - HbA1C
(Glycosylated Hb)

Light Blue Sodium Citrate Ca+2 chelating agent - PT: Prothrombin

Time
- PTT: Partial
Thromboplastin Time
( in case of
unexplained bleeding
and liver disease)
Green Sodium Heparin binds to Thrombin and inhibits Enzymes
Heparin or the second step in the coagulation Hormones
Lithium cascade Electrolytes (Na+, K+,
Heparin (Prothrombin Thrombin)
Heparin Mg+, Cl-

Fibrinogen Fibrin
Top Color Additives Principle Uses
Black Sodium Citrate Ca+2 chelating ESR ( Erythrocyte Sedimentation
agent Rate)
to test how much inflammation in
the patient, unexplained fever,
Arthritis, Autoimmune Disorder
Gray -Sodium Fluoride Glycolysis Glucose tests
inhibitor
-Potassium Oxalate Anti-Coagulant
Royal Blue Heparin Anti-Coagulant Toxicology
Na-EDTA Tube should not Trace Elements and metals
be
contaminated
with metals
Yellow ACD ( Acid-Citrate Anti-Coagulant DNA Studies
Dextrose) Paternity Test
HLA Tissue Typing
(Human Leukocyte Antigen)
The body used this protein to
differentiate the self-cells from non-
self cells
 Serum Separating Tubes (SST)

Top Tubes Additives Principle Uses

Red ------ Enhancing the Serology
Sometimes it has formation of blood -Antibodies
gel or silicon at the clot -Hormones
bottom of tube to -Drugs
reduce hemolysis Virology
Chemistry
Blood cross
matching before
blood transfusion
Gold ------- Serum separating Serology
It has gel at the from the blood Chemistry
bottom of the tube through the gel in
to separate serum the tube
from the blood