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 physician 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.
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.
 PHLEBOTOMY
Phlebotomy or blood collection:
The term phlebotomy refers to blood
draw from a vein, artery, or the capillary
bed for lab analysis or blood transfusion.

The phlebotomy equipments:

For specimen collection, the following
materials will be required:
 SELECTING VEIN SITE
Usually vein is used to collect blood by
veinpuncture procedure.
In adults: most venipuncture procedure use
arm vein.
On arm, one of three arm veins is used:
median cubital vein "located on the
middle", cephalic vein or basilic vein
"located on both sides".
Median cubital vein is the best choice
(why?) because it has good blood flow than
cephalic and basilica which has slower
blood flow.
However if veinpuncture procedure is
unsuccessful in median capital; cephalic or
basilica is used.
Artery blood is rarely used in special
cases as when blood gases, pH, PCO2,
PO2 and bicarbonate is requested. It is
usually performed by physicians.
 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 continuo which will alter
analytes conc. like glucose and lactate.
SERUM
Second 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.
PROCEDURE OF SERUM PREPARATION

• Draw blood from patient. Select vacutainer with no anticoagulant.

• Allow to stand for 20-30min for clot formation.
• Centrifuge the sample to speed separation and affect a greater
packing of cells. Clot and cells will separate from clean serum
and settle to the bottom of the vessel.
• The supernatant is the serum which can be now collected by
• Dropper or pipette for testing purposes or stored (-20°C to -
80°C) for subsequent analysis or use.
PLASMA
Third Plasma:
• The tube will have anti-coagulation
• After centrifugation the blood sample got
separated into three layers
PROCEDURE OF PLASMA PREPARATION

• Draw blood from patient. Select vacutainer with an appropriate

anticoagulant.
• Mix well with anticoagulant.
• Allow to stand for 10min.
• Centrifuge the sample to speed separation and affect a greater
packing of cells.
• The supernatant is the plasma which can be now collected for
testing
• Purposes or stored (-20°C to -80°C) for subsequent analysis or
use.
 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
Hemolysis :
• It means liberation of hemoglobin due to rupture of RBCs.
• Due to hemolysis plasma or serum appears pink to red color.
• It causes elevation in: K+, Ca2+, phosphate, SGOT, SLDH and acid phosphatase.
• 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.
BLOOD COLLECTION TUBES:
BLOOD COLLECTION TUBES:

Two major types of blood collecting tubes:

• Serum separating tubes (SST)
• Plasma separating tubes (PST)
 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 Heparin binds to Thrombin and inhibits Enzymes
or Lithium the second step in the coagulation Hormones
Heparin cascade Electrolytes (Na+, K+,
(Prothrombin Thrombin) Mg+, Cl-
Heparin
Fibrinogen Fibrin
Top Color Additives Principle Uses
Black Sodium Citrate Ca+2 chelating ESR ( Erythrocyte Sedimentation Rate)
agent 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 gel formation of blood -Antibodies
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 to through the gel in
separate serum from the tube
the blood
 Sterilization : Sterilization is the process of destruction or elimination of
all forms of microorganisms by physical means (including heat, radiation,
and filtration) and chemical agents (acids, alkalis, heavy metals, salts,
halogens, etc). The equipment or material treated then becomes “Sterile”.

• In medical laboratories, materials and equipments are sterilized for the following
main purposes. 1. In preparation for taking specimens, such as needles, syringes, test
tubes, etc. 2. To sterilize contaminated materials and equipments. 3. To prepare
apparatus used for bacteriological cultures, such as petridishes, Pasteur pipettes, and
others. Methods of sterilization techniques. A. Physical method 1. Dry heat (hot air
oven, flaming and red - hot) 2. Moist heat (autoclave or steam under pressure and
boiling 3. Radiation.
• Steam under pressure (autoclave) Autoclave is an instrument that operates by
creating high temperature under steam pressure. Autoclaving is the most common,
effective, reliable and practical method of sterilizing laboratory materials.
• The principle of autoclave is that steam is kept at a pressure of 15 pound (lb) per
square inch to give a temperature of 121 oc, which will kill spores with in 15 minutes.
At this particular temperature, pressure and time, all forms of lives are destroyed.
• Disinfection Disinfection is the process of destruction of pathogenic or potentially
pathogenic microorganisms by chemical means. All specimens, cultures and other
material that have been examined in a laboratory must be made non-infectious before
being discarded or leaving the laboratory. This is necessary to protect all those health
professionals working in the laboratory and the members of the general public who
may handle the waste materials before its final disposal. Ideally all material should be
sterilized. Unfortunately it is not always possible to achieve this. But the most
recommended method is the destruction of the vegetative forms of all pathogenic
organisms. Laboratory waste, which includes articles that will be reused, may be
disinfected by chemical or physical means.
• A. Phenolic compounds B.Chlorine (sodium hypo chloride)
• C Alcohol Ethanol and isopropanol, at 70 -80% volume / volume (v/v) concentration
in water, are useful for skin, work surfaces of laboratory benches and biosafety
cabinets, and to soak smallsurgical instruments. They are active against
vegetativebacteria, fungi and lipid containing viruses, but not against spores. Their
activity is improved by mixing them with formalin or hypochlorite. A major advantage
of aqueous solutions o falcohols is that they do not leave any residue on treated
• D. Formaldehyde E. Glutaraldehyde
• F. Iodine and iodophors
• Classification of Clinical Laboratories According to WHO
• 1. Biosafety Levels (BSL)
• The WHO classifies laboratories based on their biosafety levels to ensure
appropriate containment and safety measures when handling infectious
agents. This classification is essential for protecting laboratory personnel,
the environment, and the public.
• Biosafety Level 1 (BSL-1)
• Risk Group: Organisms not associated with disease in healthy adult
humans.

• Examples: Non-pathogenic strains of Escherichia coli, Bacillus subtilis.

• Containment Requirements: Standard microbiological practices; no
special containment equipment required.
• Personal Protective Equipment (PPE): Laboratory coats, gloves, and eye
protection as needed
• Biosafety Level 2 (BSL-2)
• Risk Group: Agents that pose moderate hazards to personnel and the
environment.
• Examples: Staphylococcus aureus, Salmonella spp.,
• Containment Requirements: Biological safety cabinets (Class I or II) for
procedures that may produce aerosols; autoclave available for
decontamination.
• PPE: Laboratory coats, gloves, face protection as needed.
• Biosafety Level 3 (BSL-3)
• Risk Group: Indigenous or exotic agents that may cause serious or potentially
lethal disease through inhalation.
• Examples: Mycobacterium tuberculosis, SARS-CoV-2.
• Containment Requirements: Controlled access; decontamination of all waste;
physical separation from access corridors; specialized ventilation systems.
• PPE: Laboratory coats, gloves, respiratory protection (e.g., N95 respirators),
and other specialized equipment
• Biosafety Level 4 (BSL-4)
• Risk Group: Dangerous and exotic agents that pose a high individual risk of
aerosol-transmitted laboratory infections and life-threatening disease.
• Examples: Ebola virus, Marburg virus.
• Containment Requirements: One-piece positive pressure suit with a dedicated air
supply; laboratory located in a separate building or isolated zone; strict entry and
exit procedures.
• PPE: Full-body, air-supplied positive-pressure suits.
• 2. Laboratory Functions and Classifications
• In addition to biosafety levels, WHO classifies laboratories based on their
functions and roles within the healthcare system.
• Primary Laboratories
• Services Provided: Basic diagnostic tests such as urinalysis, fecalysis, blood
typing, and routine hematology.
• Purpose: Serve as the first point of contact for laboratory services; essential for
primary healthcare settings
• Secondary Laboratories
• Services Provided: All services of primary laboratories plus additional tests like
routine clinical chemistry, Gram staining, and cross-matching.
• Purpose: Provide more specialized testing; often serve as referral centers for primary
laboratories.
• Tertiary Laboratories
• Services Provided: All services of secondary laboratories plus advanced tests such
as special chemistry procedures, special hematology (e.g., coagulation studies),
immunology, and culture and sensitivity procedures.
• Purpose: Offer specialized diagnostic services; often affiliated with academic or
research institutions.
• 3. Laboratory Networks
• WHO also recognizes the importance of laboratory networks in strengthening health
systems, especially in resource-limited settings. These networks ensure quality
assurance, standardization, and efficient referral systems.
• Peripheral Laboratories: Located at the community or district level; provide basic
diagnostic services.
• Intermediate Laboratories: Located at the regional or provincial level; offer more
specialized services and support peripheral laboratories.
• National Reference Laboratories: Located at the national level; oversee quality
control, provide specialized testing, and support the entire laboratory network
• Conclusion
• Understanding the WHO classification of clinical laboratories is crucial for ensuring
appropriate safety measures, quality of diagnostic services, and effective healthcare
delivery. This classification aids in resource allocation, training, and the
establishment of standardized protocols across different levels of healthcare facilities.
• .
Classification Basis Categories/Levels

Biosafety Level BSL-1, BSL-2, BSL-3, BSL-4

Function Primary, Secondary, Tertiary

Peripheral, Intermediate, National

Laboratory Network Level
Reference