Urine:

Urine specimens are either random or timed. A random sample is collected at a time that had not been
designated beforehand. Random urine samples vary significantly from each other.

A timed specimen is collected within a specified time range. e.g. a 24-hour urine sample is collected over
24 hours: timing is started immediately after the bladder is emptied and the resultant urine sample is
discarded; thereafter, all urine should be collected and stored until the end of the scheduled period.
Timing is stopped immediately after the bladder is emptied and the resultant sample is collected and
mixed with the already collected urine sample.

Faeces:

It is collected into a clean container and used for parasitological work. It is collected from the rectum of
paediatric patients with the use of a thick-walled glass tube.

Other specimens:

These include sputum, cerebrospinal fluid, tears, sweat, saliva and fluids that are obtained from the
body cavities (pleural fluid, synovial fluid).

THE PROCESSSING OF SPECIMENS

It involves the chain of events (that consist of 3 phases) that occur between specimen collection
and the laboratory analyses of the specimens.

(a) Pre-centrifugation phase:

Specimens should be received by the clinical laboratory staff within 45 minutes of their collection. When
the specimen is in transit, it should not become contaminated by getting it in contact with the tube
stoppers; it should not be agitated so as to avoid haemolysis (the breakdown of red blood cells); it
should not be exposed to light so as to prevent the breakdown of analytes (substances whose levels are
to be measured) such as bilirubin, which are light-sensitive. Samples with unstable analytes such as NH3
should be transported on ice. Specimens that would be used to measure the blood gases should be in
sealed containers that are in an ice bath; such a specimen should be discarded if it contains clots and air
bubbles. Specimens that are to be mailed in the frozen state are packed in a styrofoam container that
contains dry ice at -70oC. All assays (measurements) should ideally be carried out within 1-2 hour after
the collection of the specimen. If this is not possible, the specimen should be processed such that it can
be stored.

(b) Centrifugation phase:

A centrifuge generates a centrifugal force as a result of the rotation of its rotor (centrifuge head). This
leads to the separation of substances that differ significantly in their masses.

The relative centrifugal force (RCF) = krn2, where k is a constant that has a magnitude of 1.118 ×
10-5; r is the horizontal distance from the centre of rotation to he bottom of the tube in the rotor cavity
during centrifugation; n is the speed of rotation of the rotor.

The rotor must be balanced to allow for the proper functioning of the centrifuge. This is
achieved by ensuring that the same weights are placed on opposite sides of the rotor.

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 As part of quality control, the centrifuge has to be regularly calibrated.e.g. under specified
conditions, the measured speed of rotation should not differ by 5% from the speed that has been stated
by the manufacturer. To calibrate an instrument means to adjust it such that it can be used to accurately
measure a given parameter.

NB: The centrifugal force is expressed as a multiple of the gravitational force.

(c) Post-centrifugation phase:

Refrigeration of the processed specimen at 0oC or 4oC is an effective way in retarding its degradation
(due to the activity of enzymes in the sample; due to microbial contamination of the sample or the
spontaneous breakdown of analytes in the sample.

PERFORMING OF LABORATORY ASSAYS:

The reliability of the data from the clinical laboratory is influenced by the reagent type (water
that is contaminated with cations will lead to the generation of incorrect values of enzyme levels) and
the proper management of laboratory equipment.

(a) Reagents: They exist in different grades or degrees of purity. Pure chemicals (“reagent grade”)
should be used in quantitative analysis and the preparation of standard solutions.
(b) Water: Pure or “reagent grade” water is obtained by distillation and de-ionization of impure
water. Sterile water (that is free of micro-organisms) is obtained by the pore filtration of de-
ionized water.
(c) Measurement of mass: This is crucial in every analysis and is done by the use of the balance. The
balance must be kept clean and free from vibrations.
(d) Temperature control: This is achieved by using constant temperature water baths or heating
blocks.
(e) Glassware: Glassware should have a high thermal resistance, a low alkali content and should be
free of heavy metals. Glassware that is made up of borosilicate would be dissolved by alkaline
solutions.
Glassware for laboratory use should be cleaned in the following way: rinse with tap water; soak
in a weak detergent; wash and then rinse with tap water. Glassware with stubborn stains should
be soaked overnight in H2SO4/dichromate solution and then rinsed with tap water and cleaned
as outlined above. Glassware that has been used for bacteriological analysis should be soaked in
an antiseptic solution, autoclaved and then washed.

INFLAMMATION

This is a local response to tissue damage. This damage can be induced by:

(i) physical agents. e.g. excessive heating or cooling, mechanical trauma such as cutting or crushing.

(ii) chemical agents

(iii) living organisms.

(iv) antigen-antibody interactions

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(v) necrosis (death of tissue) would induce inflammation in the surrounding tissue.

Acute inflammation

It is caused by short—term tissue damage, and it involves the following processes:

(1) Active hyyperaemia:

Tissue damage is immediately followed by a transient constriction of arterioles. This is followed

by arteriolar relaxation in and around the damaged tissue, leading to hyperaemia. This accounts for the
redness of the damaged tissue. This increased flow of blood to the damaged tissue warms it up.

(2) Exudation:

(a) Exudation of fluid: Protein-rich fluid escapes from blood vessels (venules and capillaries in the zone
of injury) into the surrounding tissue fluid. The movement of plasma proteins into the tissue fluid
promotes the osmotic movement of water in the same direction. Exudation of fluid leads to oedema
(the swelling of the inflamed tissue).

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