dr. tarek el sewedy. clinical laboratory classification balances and centrifuges
TRANSCRIPT
PHAROS UNIVERSITY
FACULTY OF ALLIED MEDICAL SCIENCE
CLINICAL LABORATORY
INSTRUMENTATION
(MELI-201)
Dr. Tarek El Sewedy
LECTURE 2
Clinical Laboratory classification
Balances and Centrifuges
Intended Learning Outcomes
By the end of this lecture the student would learn the following:
1. Laboratory classification.
2. Balances.
3. Centrifuges
Lecture content
1. Laboratory classification.
2. Balances.
3. Centrifuges
EARLY LABORATORY INSTRUMENTATION
EARLY LABORATORY INSTRUMENTATION
EARLY LABORATORY INSTRUMENTATION
EARLY LABORATORY INSTRUMENTATION
Classification of clinical laboratories
The world Health Organization (WHO) lists four
kinds of levels of laboratories based on their
biosafety.
1. Basic laboratory level I
2. Basic laboratory level II
3. Containment laboratory (Level III)
4. Maximum containment laboratory
Basic laboratory level I
Basic laboratory level I is the simplest kind and adequate for
work with organisms which have low risk to the individual
laboratory personnel as well as to the members of the
community.
Such organisms are categorized under Risk Group I by WHO.
These organisms are unlikely to cause human diseases.
Example, food spoilage bacteria, yeasts.
Basic laboratory level II
Basic laboratory level II is suitable for work with organisms that
predispose to moderate risk to the laboratory worker and a limited
risk to the members of the community.
Such organisms are categorized under Risk Group II by WHO.
They can cause serious human diseases but not serious hazards due
to the availability of effective preventive measures and treatment.
Example, staphylococci and streptococci.
Such laboratory should be clean, provide enough space, have
adequate sanitary facilities and equipped with autoclave.
Level III Containment laboratory
Containment laboratory is more advanced and it is used for work
with infectious organisms that present a high risk to the laboratory
personnel but a lower risk to the community.
Such organisms are categorized under Risk Group III by WHO.
Example, Salmonella and HIV.
The principle is to isolate those organisms from the basic laboratory
and limit the activities which are particularly hazardous. They are
easily transmitted through airborne, ingestion of contaminated food
or water.
Such laboratory should be a separate room with controlled access
and clothing by authorized staff. It should also be fitted with
microbial safety cabinet.
Maximum containment laboratory
Maximum containment laboratory is intended for work with
viruses, which predispose to a high risk for both laboratory
personnel and the community.
Such organisms are categorized under Risk Group IV by WHO.
Example, Small pox, Ebola and others.
Most of these organisms cause serious disease and readily
transmitted from on person to another. These laboratories are
usually a separate building with strictly controlled access.
Respect Lab Containment rules for your safety
Balances
Balances
Balances in medical laboratory may be:
A. Rough balances (mechanical
balances)
B. Analytical balances
should be kept clean and located in an area
away from heavy traffic, large pieces of
electrical equipment, and open windows.
To minimize any vibration, as interference
that may happen, a slab of marble is
placed under the balance.
Rough balances
• Used for weighing substances, which do not call for extreme
accuracy.
• They do not require electricity or battery power.
used to:
• To weigh large amounts (up to several kilo grams)
• When a high degree of accuracy is not required.
Analytical balances
Highly sensitive instrument.
requires electricity or battery
These balances are used:
1. To weigh small quantities usually in milli gram(mg)
range.
2. When great accuracy is required.
Use and care of balances
Read carefully the manufacturer’s instructions.
Position the balance on a firm bench away from vibration.
Before starting to weigh, zero the balance as directed by
the manufacturer.
Never put the chemicals directly on the balance pan.
Use small brush to remove any chemical, which may have
been spilt on the balance.
Container of silica should be kept inside the balance case to
remove any moisture present in the atmosphere.
Keep the balance clean, being particularly careful not to let dirt
accumulate near the pivots and bearings.
Centrifuges
Centrifuges
Is an equipment that is used to separate solid matter from a liquid
suspension by means of centrifugal force.
They sediment particles (cells, bacteria, parasites, etc.) suspended in fluid
The suspended materials are deposited in the order of their weight.
Different materials need a different rate of spinning to obtain adequate
separation. So, all centrifuges can be adjusted to control the rate
at which the sample spins.
Spinning can be measured as revolutions per minute (rpm)
Basic principle of operation
When a body is rotated in circular movement at
speed, centrifugal force is created that drives the
body away from the center of the circular movement.
The greater the outward pull due to rotation
“centrifugal force”, the more rapid and effective is
the sedimentation. As a result, elements are
thrown to the bottom of the tube followed by
particles.
heavier
lighter
Centrifugal force increases with the speed
of rotation that is the revolution of the
rotor per minute (rpm) and the radius of
rotation.
The actual sedimentation achieved at a
given speed depends therefore, on the
radius of the centrifuge.
Classes of centrifuges and their applications
Differential centrifugation.
Separation is achieved primarily based on the size and weight of the
particles in differential centrifugation. This type of separation is
commonly used in simple pelleting and in obtaining partially-pure
preparation of subcellular organelles and macromolecules.
For the study of subcellular organelles, tissue or cells are first disrupted to
release their internal contents. This crude disrupted cell mixture is
referred to as a homogenate. During centrifugation of a cell homogenate,
bigger and heavier particles sediment than lighter ones and this
provides the basis for obtaining crude organelle fractions by differential
centrifugation.
FASTER
Differential centrifugation of a tissue homogenate
1000g/10 min
Decant supernatant
3000g/10 minetc.
Basic components of centrifuges
1. Shaft: - It is the central part that rotates when
spinning is effected.
2. Rotor: - It is a part that holds the bucket
3. Bucket: - Are portions that hold test tubes
containing a given sample.
Special centrifuges called ultracentrifuges can exceed
100,000 rpm.
The forces in a sample spinning at 10,000 rpm can
exceed 17,000 g-force units. This is the equivalent of a
150-pound person being pressed upon by a 1,275- kg
weight.
Ultracentrifuges can exceed 1 million g-force units.
Respect the Ultra
CsCl Gradient UltracentrifugationSeparation According to density
Cont , gradient centrifugation
Hand centrifuges:
Operated by hand and they are most commonly used
in small laboratory for routine purposes, Used for
preparation of urinary sediments and to concentrate
parasites from the given specimen.
Rotors Based on their tube angle rotation, there are two types.
A. Swing head: -
The head is designed to swing the tubes to the
horizontal position during centrifugation process.
B. Fixed head: -
They are fixed and have different angles.
Use and care of centrifuges
1. Read the manufacturer’s instructions.
2. Placing a centrifuge on a firm level bench
3. Whenever possible using plastic tubes made from
polystyrene or autoclavable.
4. Always balancing the tubes that are being centrifuged.
5. Never open the centrifuge while it is still spinning. Never try
to slow it down with your hand.
6. Most centrifuges have a brake, use it only when appropriate
Assignment
Assem Talal: is selected to make the
assignment on different types of
centrifugation.
Suggesting reading
Encyclopedia of Medical Devices and Instrumentation, 2nd ed. New York: Wiley, 2006