Clinical Laboratory Methods

for year II Comprehensive

Nurse

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 CHAPTER ONE

Introduction
to Clinical Laboratory Methods

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 Outline
1.1 Definition & organization of Medical Lab.
1.2 Role of Clinical laboratory
1.3 Selection, evaluation and interpretation
of laboratory tests
1.4 Laboratory equipments and wares

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 Introduction
1.1 Definition and organization of Medical
Laboratory
Laboratory
 is a place that is equipped with different
instruments, equipments, chemicals etc for
performing either experimental works or
research activities.

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 Introduction…
Medical laboratory
 is part of the laboratories that is equipped
with various biomedical instruments and
chemicals for performing different lab
diagnostic activities by using biological
specimens. (Whole blood, serum, plasma,
urine, stool, sputum etc). It is also called
clinical laboratory science .
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 Intro ….
Medical laboratory Technology
 is the health profession that provides
laboratory information and services needed
for the diagnosis and treatment of disease.

Clinical laboratory method

 is methods that can measure the amount of
clinical substances or detects and identifies
micro organisms in a given clinical sample.

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 Intro ….
 Clinical Laboratory Methods
• is a discipline that could be related with all
fields such as;
1) Hematology 5) Microbiology
2) Serology 6) Urinalysis
3) Immunohematology 7) Clinical Chemistry
4) Parasitology etc…..

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 Intro ….
Hematology
– It is a branch of Medical science and a modern
discipline that dealing with the formation,
composition, Function and diseases of blood, and
also it’s associated with diagnostic laboratory
Procedures and morphological interpretation of
blood.
Clinical chemistry
– it is a science that deals with observation and
investigation of biochemical change in human body.
Therefore, it involves the detection and
measurement of biochemical constituent (analytes)
in the given body fluids.
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 Intro ….
Immunohematology (blood bank)
• Is the area of laboratory medicine dealing with the
general procedures involved in collecting,
preparing, storing and transfusing blood.
Urinalysis
• deals with the analysis of urine constituents.
• Consists of physical, chemical and microscopic
examination
Serology
• is the study of antigen and antibody and their
interaction in vitro.
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 Intro ….
Medical microbiology
– deals with microorganisms of medically important,
performing different diagnostic and research
activities on microorganisms and parasites.

Sub-division of Medical Microbiology

 Medical Bacteriology
 Medical Virology
 Medical Mycology
 Medical Parasitology
 Immunology
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 Summary of laboratory service network

central or National
Reference Laboratory

regional regional
laboratories laboratories

District or District or District or District or District or District or

hospital hospital hospital hospital hospital hospital
laboratories laboratories laboratories laboratories laboratories laboratories

Health center Health center health center health center health center health center
laboratories laboratories laboratories laboratories laboratories laboratories

•Note:- There are also private hospital, clinic& diagnostic

laboratories 11
 Structure of lab. service
Types of laboratory
• Structure and function of lab. Service in developing
countries.
A) Community based primary health care lab.
B) District hospital lab
C) Regional (Referral hospital) laboratories
D) Central and public health lab.

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 Structure of lab…..
A. community-based health center lab:-
– Is to support health care workers by
investigation controlling and preventing major
disease in a community.
– Able to perform basic tests and
– refer specimen to district hospital
• Note:- In Ethiopia these laboratories are called zonal
or district and health center laboratories
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 Structure of lab…..
B. District Hospital laboratories (DHL)
 In addition to the work stated above the laboratories
have an important role in:
• To perform a range of tests relevant to the medical,
surgical, and public health activities of the district
hospital.
• Supervising the work of community-based health
center lab.
• Testing referred specimens
• refer specimens to the regional lab
• To participate in the external quality assurance
program organized by the regional lab.
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 Structure of lab…..
C. Regional (Referral hospital) laboratories
• In addition to the work stated above the
laboratories
– It assists and supervise the DHL.
– Analyses referred samples .
– Perform a range of specialized testes.
– Investigate epidemic and do more clinical and
public health facilities.
– Trains lab technicians by organizing refresher
courses.
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 Structure of lab…..
D. Central and public health laboratories
– This lab is responsible for the planning,
expenditures, and co-ordination of the national
lab service.
– It has equally important roles in ensuring the
reliability of the service.
– The appropriateness of its technology, training
and motivation of its work force, and insuring that
the service extends into the area of health needs
and that its facilities are made available to as
many people as possible.
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 1.2 Role of the clinical laboratory
• Role of the clinical laboratory in the health care
systems:-
 Patient : - Diagnosis
- Screening and
- Monitor (treatment follow up)
 Public health aspects such as:
• Epidemic control
• Surveillance and
• Delivery of health information

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 Role of the clinical lab......
 Medical Laboratory and Its Function in a Health
Care System
• Laboratory has an important role in:
1) Providing quality health care service
2) Achieving efficiency and cost effectiveness
in health care system.
3) Achieving good health planning and
management in health care system.

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 Role of lab……………
1) The role of laboratory in providing quality
health care
a) laboratory investigation increase the
accuracy of disease diagnosis.

 Many infectious disease and serious illness

can be diagnosed by using the laboratory.
 E.g. error in diagnosis of malaria is high
when diagnosis is on clinical symptoms only.
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 Role of lab………………….
b) Laboratory has essential role in screening for ill
health and assessing response to treatment.

 Assess a patients response to drug therapy.

 Screen individuals with infectious disease. E.g.
TB, sexually transmitted diseases.
 Screen whole blood and blood product for
transfusion transmitted pathogen .
 Screen pregnant women for anemia, infections
which may transmitted to newborn like syphilis
and others.
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 Role of lab………………….
c) The laboratory is needed to work with other
in reducing infection in the community and
investigating epidemics rapidly.
• Detecting source of infection, identifying
carries contact tracing
• Participating in epidemiological surveys
• On site testing and collection and dispatch
of specimens when an epidemic occurs
• Participating in health education
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 Role of lab………………….
2) The role of laboratory in achieving efficiency
and cost effectiveness in health care system.

i) The laboratory reduce the expenditure of

money on drugs.
 Drugs can be used more selectively and only
when needed.
 Drug resistance can be identified more rapidly
and monitored.

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 Role of lab………………….
ii) Laboratory lowers health care costs by
identifying disease at an early stage.
Reduce number of times a patient may need
to seek medical care for same illness.
Prevent complications
Avoid hospitalization and further costly
investigation

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 Role of lab………………….
iii) Saving can be made when laboratory
participate in local disease surveillance and
control ,this is because

source of infection and disease carriers can

be identified.
the spread of infectious disease can be
controlled
disease control measures can be selected
and targeted more effectively.

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 Role of lab………………….
3) The role of laboratory in achieving good health
planning and management
a) Reliable laboratory test result with relevant patient
data, provide information on the health status of a
community, health pattern and disease trend. This
information is needed to establish health care
priority and plan:
 Health care program.
 Training of health personnel and delivery of health
service.
 Treatment schedule and change in drug usage.
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 Role of lab………………….

b) Public heath laboratories provide accurate

epidemiological information for health
planning
– cause of the disease in the community.
– prevalence and incidence rate of infectious
disease.

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 Role of lab………………….
Importance of requesting lab examinations
To make a diagnosis of pathogen.
Confirm a clinical impressions.
To rule out (R/o)a disease and diagnosis.
To provide prognostic information.
To screen for disease.
To provide therapeutic guidance so that:
• Drugs can be used more selectively.
• Patterns of emerging drug resistance can be
identified.
• Side effects and progress are monitored.
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1.3 Selection, evaluation and interpretation
of laboratory tests
Selection of laboratory tests depends on
 The value of the information they provide

 Cost of the test

 Availability of the necessary laboratory materials

 Availability of well trained & experienced lab

staff.

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 Interpretation of Lab. test results

• To interpret the data needs to answer the

following question :
Is the result normal or abnormal according to
reference range.
Does the result fit with my assessment ?
Does a significant change occurred in the result
?with previous test result/reference range.
Do the result alter my diagnosis of the patient ?
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 Evaluation and interpretation of
laboratory tests
 Precision & accuracy
• For effective diagnosis & management of
patients, a method with high precision & accuracy
must be used.
– Accuracy:
• is the closeness of measurements to the
true value
– Precision:
• the closeness of repeated measurement to
each other
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 Accuracy and Precision
Accurate Precise
and Precise Imprecise

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 Quality
• The ability of a diagnostic test to indicate whether a
disease is present or absent is dependent on its
quality.
• Quality is described in terms of:
• Sensitivity
• Specificity
• Predictive value
–PPV test
–NPV test
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 Quality ……
Sensitivity
– The ability of the diagnostic test to detect
very small amount of the analytic
substance.
– is the ability of a test to identify correctly
those who have the disease.
– Identify true positive.

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 Quality ……
Specificity
• is the ability of a method to:
– identify all samples, which do not contain the
substance being detected or
– is the ability of a test to identify correctly those who
do not have the disease
– identify true negative.

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 Quality ……
Predictive value (PV)
– Predictive value is the ability of a test to predict the
presence or absence of disease from test results.
– PPV and NPV
 Positive Predict Value (PPV)
• is the probability that an individual with a positive
test result has the disease.
• the proportion of diseased individual out of positive
test.

PPV = diseased individual by the test x 100

the whole positive test
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 Quality ……
• Negative Positive Value (NPV)
– is the probability that an individual with a
negative test result does not have the disease.
– the proportion of not having the disease when
the test is negative.

PPV = non-diseased individual x 100

the whole negative test

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Screening Golden standard (diseased ) Total
test Yes No
Positive A B A+B
True positive False positive
Negative C D C+D
False Negative True negative
Total A+C B+D A+B+C+D
The whole disease The whole non-
disease

 Sensitivity =__A__ = Test⁺ PV⁺= _A_ = Dx⁺

A+C Dx⁺ A+B Test⁺

Specificity=__D__ = Test⁻  PV⁻ =_D___ = Dx⁻

B+D Dx⁻ C+D Test⁻
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 Calculating Sensitivity, Specificity, PPV, & NPV

Actual HIV status (Gold Standard)

Test
result HIV infected HIV -uninfected Total
Positive A (370) B (2) A+B(372)
Negative C (4) D (624) C+D(628)
Total A+C (374) B+D (626) 1000

Sensitivity = A ÷ (A+C) = 370 ÷ 374 = 98.9%

Specificity = D ÷ (B+D) = 624 ÷ 626 = 99.7%
PPV = A ÷ (A+B) = 370 ÷ 372 = 99.5%
NPV = D ÷ (C+D) = 624 ÷ 628 = 99.4%
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 1.4 Laboratory equipments and wares
Laboratory equipments
1. Microscope
• It is an instrument used to magnify objects that are not
visible to our naked eye.

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 Lab. equipments…..
Working principle of a microscope
• A microscope is a magnifying instrument. The
magnified image of the object (specimen) is
first produced by a lens close to the object
(objective). This collects light from the
specimen and forms the primary image. A
second lens near the eye called the eye piece
enlarges the primary image, converting it in to
one that can enter the pupil of the eye.

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 Lab. equipments…..
Types of microscope
A. Based on the number of lenses they use:
• Simple microscope: Ordinary magnifying
lens is known as simple microscope. It has
5x, 10x, and 20x magnification.
• Compound microscope: a system of lenses
used. It has 40x, 100x, 400x and 1000x
magnification.

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 Lab. equipments…..
B. Based on the principle they use:
1) Light (Optical) microscope:
• use a system of optical lenses and light waves to
produce the image of the object.
The types of light microscope include:
i) Bright – Field Microscope
ii) Dark – Field Microscope
iii) Fluorescence Microscope
iv) Phase – Contrast Microscope

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 Lab. equipments…..
2) Electron microscope:
• uses a beam of electrons produced by an
electron gun to produce the magnified image.
• Mainly used in
– Negative staining
– Examination of viruses
NB. The beam can not pass through the metallic
back ground of the microscope.

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 Lab. equipments…..
C. Based on the source of light they use:

– Those using sun light

– Those using electricity

D. Based on the number of oculars (eye pieces):

– Monocular Microscope

– Binocular Microscope

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 Lab. equipments…..
 Structural categories of compound microscope
1) The frame work 3) The magnification
• Base system
• Arm – Ocular (eye piece)
• Stage – Objectives (LP, HP,
2) The illumination
system OI objectives)
• Light source 4) The focusing system
• Intensity control – Fine adjustment
• Condenser – Coarse adjustment
• Iris diaphragm – Condenser
• Mirror adjustment
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 Parts of microscope
1) Frame work of the microscope
 An arm (stand): - The basic frame of the
microscope to which the base, body and
stage are attached.
 A stage: - the table of the microscope
where the slide or specimen is placed.
 A foot or base: - is the rectangular part up
on which the whole instruments rest.

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 Part of mic…..
2. Focusing system
 Course adjustment: - The course focusing adjustment is
controlled by a pair of large knobs positioned one on
each side of the body. Give rough image.
 Fine adjustment: - it moves the stage so slowly that and
give clear image .
 Condenser adjustments: - The condenser is focused
usually by rotating a knob to one side of it.
• This moves the condenser up or down.
• The condenser aperture is adjusted by the iris
diaphragm, which is found just below the condenser.
• The principal purpose of the condenser is to
condense the light required for visualization
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 Part of mic…..
3) Magnification system
– Objectives: - Objectives are components that
magnify the image of the specimen to form the
primary image.
• For most routine laboratory work 10x, 40x and
100x (oil immersion) objectives are adequate.
– Eyepiece:- Eyepiece is the upper optical component
that further magnifies the primary image and
brings the light rays to a focus at the eye point.
• It consists of two lenses mounted at the correct distance.
• It is available in a range of magnifications usually of 10x,
15x and sometimes as high as 20x.
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 Part of mic……
4) illumination system
– Condenser :- is a large lens with an iris diaphragm.
• The condenser lens receives a beam from the light
source and passes it into the objective.
– The iris:- is a mechanical device mounted underneath
the Condenser and controls the amount of
light entering the condenser.
– Mirror
• Mirror is located below the condenser and iris.
• It reflects the beam of light from the light source
up wards through the iris into the condenser.
• The mirror is used to reflect ray or electrical light.
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 Part of mic…..
• Filters
Light filters are used in the microscope to:
• Reduce the intensity of light.
• Increase contrast and resolution.
• Adjust the color balance of the light to give the
best visual effect.
• Provide monochromic light.
• Absorb light.
• Transmit light of selected wavelength.
• Protect the eye from injury caused by ultra-
violet light..
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 How to use a microscope
1) Position the microscope correctly and
identify the essential parts.
2) Bring the 10x objective in to place
3) Make sure the under side of the specimen
slide and surface of the stage are completely
dry and clean.
4) Place the specimen on the stage in the slide
holder.
5) Focus the specimen with the 10x objective
6) Focus the consider 53
 how to use………….
7) Examine the specimen with the 10x objective
- Closing the iris about 2/3 and Adjusting the
lamp brightness
8) Use the mechanical stage to examine specimen
systematically.
9) Examine the specimen with the 40x objective
- Opening the iris and Increasing the
illumination
10) Examine the specimen with the 100x objective
– Place a drop of oil on the slide
– Opening the iris fully
– Increasing the illumination
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 Other lab equipments.
• Balance ( analytical and • Autoclave
mechanical) • PH meter
• Centrifuges • Desiccators
• Shakers & mixers • Colorimeter
• Water bath (photometer)
• Incubator • Oven
• Refrigerators • Distiller etc….
• Biological safety
cabinets

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Laboratory Glass wares and Plastic wares
• Based on their function generally lab. Glassware
categorized into two major parts.
A) Containers and receivers
– They are not calibrated to hold exact volume.
– On these items only two piece of information written
• Volume
• brand name
1. Beakers
2. Erlenmeyer flask
3. Test tubes
4. Reagent bottles
5. Cuvettes
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 Glass wares…..
B) Volumetric glass ware
 Calibrated to hold specific volume (volume, it going to
measure.)
 Used in the preparation of specific volume of reagent or
solution.
 Eg. of volumetric glassware
1. Graduated measuring 3.Pipettes
cylinder a) Delivery pipettes
2. Volumetric flask –  Volumetric type (bulb)
- Flat bottom  Graduated pipette
- Round bottom b) Pipette to contain
 Bulb type
- Volumetric
 Graduated pipette
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 A. B C D.

A. Volumetric (transfer) B. Ostwald folin (transfer). C. Measuring (Mohr) D. Serological (Graduated)

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A. Conical B. Flat bottomed C. Flat bottomed D.Volumetric

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 Volumetric flask

Beaker

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Petri dishes

Reagent bottles

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Dropping bottle
Funnels Cuvettes

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