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1. TECHNICAL REPORT
ON
STUDENT INDUSTRIAL WORK EXPERIENCE
SCHEME (SIWES)
UNDERTAKEN AT
DE-FAMILY HEALTH DIAGNOSTICS & RESEARCH CENTER,
GROUND FLOOR, SUITE 2, GODS’ OWN PLAZA, 5 EGBU ROAD, IN
OWERRI, IMO STATE
BY
NJOKU CHIMUANYA EMMANUELLA
MATRIC NO.: 2019/SC/12043
SUBMITTED TO
THE DEPARTMENT OF BIOLOGY FACULTY OF
BIOLOGICAL SCIENCE
ALEX EKWUEME - FEDERAL UNIVERSITY
NDUFU ALIKE, IKWO, EBONYI STATE.
P.M.B.1010
IN
PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE
AWARD OF BACHELOR OF SCIENCE IN BIOLOGY
MAY, 2023.

2. CERTIFICATION
This is to certify that this report of the Student Industrial Working Experience
Scheme (SIWES) program was carried out by Njoku Emmanuella Chimuanya, a
student of Alex-Ekwueme Federal University Ndufu-Alike, Ikwo, Ebonyi State
with Matric No.: 2019/SC/12043 at DE-FAMILY HEALTH DIAGNOSTICS &
RESEARCH CENTER, GROUND FLOOR, SUITE 2, GODS’ OWN PLAZA, 5 EGBU
ROAD, OWERRI, IMO STATE, under the Supervision of Dr. Onyekwere Amos
__________________________
Student’s Signature
NJOKU CHIMUANYA EMMANUELLA
[STUDENT]
__________________
Supervisors Signature
DR. ONYEKWERE AMOS
[SUPERVISIOR]
APPROVED BY:
DR. CLEMENTINA UWA (H.O.D BIOLOGY DEPARTMENT)
DR. ADEOLU ADEWALE IYANIWARA (I.T COORDINATOR)
i

3. DEDICATION
I dedicate this report to God Almighty; who is the giver, and sustainer of life for
preserving and keeping me even through those challenging periods of the
elections and throughout the period of my Industrial Training at De-family
Health Diagnostics and Research center. May His Name be glorified. I also
dedicate this report to my supporting parents Mr. & Mrs. J.E Njoku, and also to
my Siblings for all their support.

4. ACKNOWLEDGEMENT
I give thanks to Almighty God who gave me the gift of life, and made
everything possible for me to complete this Industrial Training.
This acknowledgement would be incomplete if I fail to express my
gratitude to my Parents Mr. &Mrs. J.E Njoku for their financial support to my
success. My appreciation also goes to Mrs. Confidence Ogechi Anyanwu, the
Managing Director of the Diagnostics and Research Facility I worked at and to
the staff of the organization for all the impacts they made to my Industrial
Training experience.

5. REPORT OVERVIEW
The Student Industrial Work Experience Scheme established by the
Federal Government of Nigeria was aimed at exposing students of higher
institution to acquire industrial skill and practical experience in their approved
course of study, and also to prepare students for the industrial work situation,
which they are likely to meet after graduation.
Therefore, this technical report is based on the experiences gained during
my five months of Industrial Training at De-Family Health Laboratory and
Research Centre in Owerri, Imo state.
The report also highlights the several sections I had the opportunity to
work in this past 5 months of training, sections which ranged from Phlebotomy,
Hematology/Immunology, Parasitology, Medical Microbiology, and Serology
Sections respectively. These sections helped me to explore and gain knowledge
on the Precautions, Rules and Regulations while working in a Medical
Laboratory.
Most importantly, this report describes the best approach to some Medical
diagnosis and analysis performed in the Laboratory and also stated the problems
encountered and some suggestion needed for the improvement of the SIWES
scheme.

6. TABLE OF CONTENTS
Title page
Certification…………………………………………………………................. i
Dedication……………………………………………………………………… ii
Acknowledgements…………………………………………………………….. iii
Report Overview………………………………………………………….......... iv
CHAPTER ONE
1.0 About the Industrial Training Fund (I.T.F)…………………………… 1-3
1.1 About SIWES (Student Industrial Training Fund)………………… 1
1.2 Scope……………………………………………………………….. 1-2
1.3 Aim and Objective of S.I.W.E.S……………………………………2-3
1.4 History and Background of DE- FAMILY HEALTH LAB…………2-3
1.5 Organizations’ Chart and Organogram……………………………..3
CHAPTER TWO
2.0 The Laboratory…………………………………………………………... 4-6
2.1 Introduction to the Laboratory…………………………………….... 4-5
2.2 Safety Rules in the Laboratory……………………………………… 5
2.3 Emergency in the Laboratory……………………………………….. 5
2.4 Hazardous Materials………………………………………………… 5
2.5 Hazardous Equipment…………………………………………...…. 5
2.6 Laboratory Equipment and their Uses……………………………… 5-6
CHAPTER THREE
3.0 The Laboratory Sections and Various Tests Performed………………..7-28
3.1 The Phlebotomy Department………………………………………… 7-11
3.2 Hematology and Immunology Section …………………….………...12-16
3.3 Serology Section………………………………………………….….. 17-19
3.4 Clinical Chemistry Section…………………………………….………19-22
3.5 Medical Microbiology & Parasitology Section……………………… 22-28
3.6 Microscopy, Culture and Sensitivity Tests …………………….…….. 28
CHAPTER FOUR
4.0 Summary, Challenges Encountered, Recommendation and
Conclusion………………………………………………………………..…. 29
4.1 Summary……………………………………………………….……... 29
4.2 Challenges Encountered……………………………………………… 29
4.3 Conclusion………………………………………………………….… 29
4.4 Recommendation…………………………………………………...… 29

7. CHAPTER ONE
1.0 ABOUT THE INDUSTRIAL TRAINING FUND (I.T.F)
In October 1971, the federal government established the Industrial Training Fund
(I.T.F). In its policy statement no.1 published in 1973, a clause was inserted dealing with
the issue of practical skills among trained professionals in the Tertiary Institutions
especially the Universities of Technology, Monotechnics, Polytechnics, Colleges of
Education and Technical Colleges.
Section 15 of the policy statement states clearly that “Great emphasis will be placed on
assisting certain products of the post-secondary school system to adapt or orientate easily
to their possible post-graduation job environments”, subsequently leading to the launch of
a scheme known as the Student’s Industrial work Experience Scheme(SIWES).
1.1 ABOUT SIWES
The S.I.W.E.S. was launched in 1973 by the Industrial Training Fund (I.T.F) as a
program designed to impart the undergraduate of the nation’s tertiary Institutions studying
various professional courses with the practical methods of performing professional
functions to real life situations on site, in the office or even the factory and how they apply
themselves mentally, intellectually and physically in relation to what they have been
taught in the classrooms Theoretically. It works with the following professional bodies to
function effectively across the tertiary institutions nationwide. These are the Nigeria
University Commission (N.U.C), National Board for Technical Education (N.B.T.E.) and
the National Commission for Colleges of Education (N.C.C.E.). Thus, equipping the
students with the necessary skills and technical knowledge to make them highly
competitive and professional individuals in the Labor Market
1.2 SCOPE
The scheme as conducted by the Industrial Training Fund (I.T.F) through their
representative liaison units and offices situated within the various states and in major cities
or towns in Nigeria with the necessary industrial rudiments needed to corroborate, practice
and then actualize the required technical knowledge. The Industrial Training experience
not only puts them in real life situations buts also exposes their practical knowledge of the
course of study, consequently perfecting this knowledge thereby producing very
competent and versatile professionals.
1.3 AIM AND OBJECTIVE OF S.I.W.E.S
The aim of S.I.W.E.S is to bridge the gap between the level of knowledge acquired in
tertiary institutions and the practical application of such knowledge in the field of work.
The Objectives are:
 To provide an avenue for students in industries of higher learning to acquire industrial
skills and experience in their course of study.
 To prepare students for the work situations they are to meet after graduation.
 To expose students to work methods and techniques in handling equipment and
machinery that may not available in the educational institution.
 To make transition from school to the world of work easier and enhance students
contact for later job placements.

8.  ,To improve student’s interpersonal relationship with others in their field.
 To prove students an opportunity to apply his/her knowledge in real work situation,
thereby bridging the gap between college work and actual practice
1.4 BACKGROUND OF DE-FAMILY HEALTH DIAGNOSTICS AND RESEARCH
CENTER
 COMPANY’S VISION
To be a leading globally recognized Medical Diagnostic service provider, by upscaling and
updating our services through research.
 COMPANY’S MISSION
To significantly provide effective Health care, through Provisions of Excellent,
Affordable, and Reliable Medical Diagnostic Services.
 CORE VALUES
 Knowledge based on Best Medical Approaches
 Trust
 Persistence
 Timely Delivery
 Customer Satisfaction
 Integrity
 Reliability
 SCOPE OF SERVICE
De-Family Health Diagnostics and Research Center provides the following services to
people:
 HEMATOLOGY
This is the branch of medical laboratory science that specializes in the diagnosis of
blood disorders and diseases.
 MEDICAL MICROBIOLOGY/PARASITOLOGY
This is the branch of medicinal laboratory science that is concerned with the
diagnosis, prevention, and treatment of infectious and parasitic diseases.
 IMMUNOLOGY
This is where tests are done to study immune systems in patients.
 CYTOLOGY
This branch of Medical Laboratory is concerned with the study of the structure and
functions of patients’ cells in relation to screening of their cancer cells, fetal
abnormalities, and pap smears for diagnosis of their ailments.
 CLINICAL CHEMISTRY
This is a branch of medical laboratory science that focuses on qualitative tests of
important compounds/Analytes/Markers, in body fluids and tissue using analytical
techniques.

9.  SEROLOGY
Charged with ensuring the safe and effective diagnostic examination of blood serum,
especially with regard to the response of the immune system to pathogens or
introduced substances.
1.5 ORGANISATION’S CHART AND ORGANOGRAM
BOARD OF DIRECTORS
MANAGING DIRECTOR
CHIEF MED LAB SCIENTIST ADMIN MANAGER
MED-LAB TECHNICIAN MED-LAB ASSISTANT RECEPTIONIST CASHIER CLEANER DRIVER
Diagram depicting the organization level at De-Family Health Diagnostics & Research
Centre

10. CHAPTER TWO
2.0 THE LABORATORY
2.1 INTRODUCTION TO THE LABORATORY
A laboratory is a facility that provides controlled conditions in which scientific
researches, experiments, and measurements, may be performed. Hence the medical
laboratory is an environment where tests are carried out on clinical specimens in other to
get information about a patient’s health.
There are three sections in the laboratory, they are; Clinical Microbiology section,
Hematology/Serology section, and Clinical Biochemistry section. The overall significance
of the laboratory diagnosis is that they guide towards the administration of the most
effective therapy so as to restore a proper health on the patient.
Laboratory safety precautions and ethics include;
2.2 SAFETY RULES IN THE LABORATORY
Every laboratory is expected to adopt a code of bio-safety principles and work practice
which should be enforced and adhere to strictly by workers and visitors. All specimens
coming into and from the laboratory are being assumed to be potentially infectious and
harmful and that is why the below precautions are ensured to be taken to avoid
contamination and laboratory hazard.
 Avoid disrupting laboratory activities you must TURN OFF all cell phones and pagers:
their use is prohibited.
 All persons in laboratories, including students, staff, and visitors, shall wear safety
glasses, goggles, or face shields at all times where potential eye hazards exist
 Eating, drinking, chewing gum, and applying cosmetics are prohibited laboratory.
 Do not store food or beverages in the same refrigerators or freezers with chemicals,
biohazards, or radioactive materials.
 Never conduct unauthorized experiments or engage in horseplay in a laboratory. Please
immediately report any unsafe behaviour to the instructor.
 Wear appropriate clothing. In particular, you must wear closed-toed shoes (i.e., NO
sandals or flip-flops!) in the laboratory. If you have a long hair, tie it back. Avoid
wearing dangling jewellery.
 Wearing an iPod, Bluetooth, or any other device that interferes with hearing is not
allowed.
 Never pipette anything by mouth.
 The work area must be kept clean and uncluttered. All chemicals should be labelled and
stored properly.
 The hazards of chemicals used should be known (e.g., corrosiveness, flammability,
reactivity, stability, and toxicity).
 Always pay attention to your surroundings and be aware of what others are doing.
Always be courteous.
 Remove contaminated gloves before touching common use devices (door knobs,
faucets, equipment); discard gloves before leaving the laboratory.

11.  Always wash hands and arms with antibacterial soap and water before leaving the
laboratory.
In conclusion, maintaining safety in the laboratory largely rest on the shoulder of
the laboratory workers. Adequate safety and good laboratory practice can be avoided
irrespective of the location, staff strength and availability of sophisticated safety cabinets
in the laboratory. What are required are highly standards of hygiene by the laboratory
workers to achieve good results in their daily occupational practice.
2.3 EMERGENCY IN THE LABORATORY
 Know where to find the nearest exit in case of fire or other emergency.
 Know the whereabouts of the nearest fire extinguisher, fire blanket, first aid kit,
eyewash equipment, shower and telephone.
 In case of fire, clear out of the laboratory first, and then call an emergency number.
2.4 HAZARDOUS MATERIALS
 Both liquid and dry chemicals can be flammable, poisonous, carcinogenic, etc. Pay
attention to special instructions, such as to; work with a substance only in a fume
hood.
 Biological hazards include bacteria and body fluids, such as blood. Handle with
appropriate care, and dispose of biological hazards as instructed.
 Dispose of hazardous materials as instructed. Never put anything down the sink
without checking with an instructor.
 Clean up spills and broken glass. Don't handle broken glass with your bare hands. Use
a broom and dustpan, and throw away all broken glass and disposable glass pipettes,
cover slips, and other sharp or easily breakable glass in a container for glass disposal
only. Notify the instructor immediately of all incidents.
2.5 HAZARDOUS EQUIPMENTS
 If appropriate, turn off equipment that isn't being used.
 Do not use a Bunsen burner unless instructed to do so.
 Keep liquids and chemicals, e
 specially flammable materials, well away from any heat source or electrical
equipment.
 If any electrical equipment is malfunctioning, making strange noises, sparking,
smoking, or smells "funny," do not attempt to shut it off or unplug it. Get an instructor
immediately. It is imperative that the instructor know of any equipment problems.
2.6 LABORATORY EQUIPMENTS AND THEIR USES
 Microscope: Is used to examine samples and to analyze their contents that are not
visible to the naked eye. It is used to count pathogen and other cells and to view under
x10, x40, and x100 objectives.
 Autoclave: For Sterilization
 Centrifuge: Is used for spinning specimen e.g. urine to enable separation into
constituents or components e.g. blood into serum and plasma.

12.  Refrigerator: Provides suitable temperature for storage and preservation of reagents,
unused media, blood samples etc.
 Biochemistry Analyzer: Used to measure parameters in tests like like Liver function
Tests, EUC, and Lipid Function Test
 Bunsen burner: Serves as the source of heat for sterilizing wire loop, surgical forceps
and other metal instruments to be used for analysis.
 Weighing Balance: Use for measurement.
 Wire loop: It is used for streaking specimen on culture plates and it can also be used for
making smear of samples on slides.
 Lancet: It is a sterile needle used to prick the thumb for the collection of blood
samples.
 Capillary tube: It is used for the collection of blood samples to determine the packed
cell volume.
 Universal bottle: used for sample collection e.g. urine, stool, semen
 Glass slide: It is used for the preparation of samples to be viewed directly under the
microscope.
 Sterile swab stick: Is used for the collection of samples to directly from the sight of
infection e.g. Ear, nose, vagina, cervix, etc.
 Sampling bottles: They are bottles used for the collection of blood samples e.g.
universal bottle, fluoride oxalate bottle, Ethylene-Di-amine-Tetra acetic Acid bottle
(EDTA), Lithium Heparin bottle, plain bottle.
 Incubator: used for culturing or drying of microorganism.
 Micro hematocrit centrifuge machine: it is used to spin sample for the analysis of
packed cell volume of blood sample.
 Water bath: Use as heating apparatus
 Micro hematocrit reader: used to read the packed cell volume in percentage.
 Tourniquet: it is tightened on patient hand in the collection of blood sample in order to
apply pressure to the prominent vein before incision.
 Needle and Syringe: It is used for the collection of blood samples.
 Macro centrifuge machine: It is used for the separation of blood samples in order to
get the plasma and also used for the separation of urine sample so as to get the
supernatant and the specimen
 Glucometer: used to check for the sugar level in the body with the aid of its strip.
 Hematology Analyzer: Is used for the analysis of Full Blood Count (FBC).

13.  Hot-air Oven: Is used for the drying, heating and often times sterilizing instruments
used in the lab.

14. CHAPTER THREE
3.0 THE LABORATORY SECTIONS AND VARIOUS TESTS PERFORMED
3.1 THE PHLEBOTOMY DEPARTMENT
The Phlebotomy procedure facilitates obtaining good quality specimens on the correct
patient for further analysis in the laboratory
3.1.1 IDENTIFICATION OF MATERIALS USED IN THE PHLEBOTOMY AND USES
1. Personal Protective Equipment
 Phlebotomy Uniform: Serve as protective to the body
 Disposable gloves/Hand Gel: For protection against spillage
2. Needles: Used to make incision
3. Needle holders: For holding needle or also known as Venoject
4. Vacutainers, Vacutainer holder or Syringe: Serves as blood drawer
5. Sample bottles according to Order of Draw:
 EDTA (Ethylene Diamine Tetra Acetic Acid)
WQ to the sample. Calcium and magnesium - these are chelated by the EDTA.
EDTA is also an unsuitable additive for samples requiring bacterial culture, since
the chelation of the divalent cations inhibits the growth of bacteria. EDTA is
sometimes used to prevent cells clumping in fluid samples requiring cell counts to
accompany a cytology evaluation but it does not actually 'fix' the cells. A sample
fixed in formalin or alcohol/ethanol is required for accurate cytology examination.
EDTA is not suitable for samples requiring virus isolation in cell/tissue culture
because it forms a gel when added to the cell culture medium and this disrupts the
cell monolayer.
 Plain (no anticoagulant)
Plain (or clotted) samples are used to provide serum for serology and most
biochemistry or endocrine assays. Serum is plasma without fibrinogen since the
fibrinogen has been used' in the formation of the clot.
 Lithium Heparin
Lithium heparin accelerates the action of anti-thrombin III which neutralizes
thrombin and thus prevents the formation of fibrin from fibrinogen (clot
formation). This effect makes heparin samples unsuitable for determination of
fibrinogen or clotting factors.
Lithium heparin is a standard anticoagulant used to obtain plasma for biochemistry
analysis. Lithium heparin is the most suitable anticoagulant for the isolation of
viruses in cell/tissue culture. This anticoagulant is not suitable for haematology as
the heparin alters the cell morphology. Whilst measurement of haemoglobin and
blood cell counts can be obtained using this anticoagulant an accurate white cell
differential and morphology comments are not possible.
 Fluoride/Oxalate
Fluoride/oxalate samples are used for glucose (and lactate) determination only.
Sodium fluoride functions by stabilizing the blood cell membrane and inhibiting

15. the enzyme systems involved in glycolysis, which prevents red blood cells
metabolising any glucose present in the sample. For this reason it is the only
suitable sample for accurate glucose analysis. Fluoride is a potent inhibitor of
many enzymes and the inhibition of glycolysis tends to cause fluid shifts. Fluoride
is a weak anticoagulant on its own, so potassium oxalate (another powerful
enzyme inhibitor) is usually added to supplement its action. Other plasma or
serum samples may be used for glucose analysis ONLY if the plasma/serum is
separated from the cells within 1 hour of sample collection. Without an anti-
glycolytic agent, the blood glucose concentration decreases by approximately 0.56
mmol/ l per hour at 25°C.
6. 70% Alcohol swabs: Used as a disinfectant for the area in which incision is to be
made.
7. Adhesive dressing
8. Rack
9. Disposable waste bins
10. Cotton wool or Pillow support: Used for supporting the arm for easy blood flow
11. Tourniquet: A transparent cord-like instrument used to apply pressure at the Veins
were blood is to be collected by tying the area
3.1.2 STANDARD OPERATION PROCEDURE (S.O.P) FOR BLOOD COLLECTION
 The frequent point of blood collection is usually from the vein (venipuncture). The
materials for the patient’s identity must be checked before attempting venepuncture.
This must be carried out by asking the patient their Full Name and Date of Birth.
 Check that this information corresponds with that on the Request form.
 Any amendment to these details or any others on the Request form must be in
accordance with the Directorate Policy.
 Where patient details lack legibility, staff may write the correct details clearly next to
those on the form without crossing out the original details.
 If tests are requested that are unfamiliar and staff are unsure of the appropriate blood
tubes for specimens check the list ‘What tube guide’ available at each workstation or,
when necessary contact a qualified Biomedical Scientist in the appropriate department
within the Pathology Laboratory.
 Examine both arms of the patient and select the one that appears appropriate
 Ensure that the patient is comfortable and that the arm is well supported and examine
potential venepuncture.
 Ask the patient to bare an arm, ensure that the arm is well supported and apply the
tourniquet to the patient’s arm, just above the elbow and tight enough to allow two
fingers behind the strap.
 Tighten the tourniquet a little more, taking care not to pinch the skin
 Ask the patient to straighten their arm and clench their fist. This will make the vein
more prominent.

16.  If necessary rub the bend of the elbow to make the vein more visible.
 Feel with a fingertip for the ‘best’ vein at the bend of the elbow rather than plunge
the needle into a poor vein that looks ‘alright’.
 If this fails a suitable vein can often be found at the side of the arm on the elbow side.
 It may be necessary, on occasion to take blood from the back of hand.
 Apply the tourniquet above the elbow. The tourniquet is closed around the arm by
inserting the plastic clip into the holder and then tightened appropriately by pulling the
strap.
 Ask the patient to straighten their arm and to make a fist in order to make the veins
more prominent.
 Feel with the fingertip if necessary to locate a suitable vein to puncture.
 Ensure that equipment and blood tubes required are immediately within easy reach.
 Remove the top plastic section of a Vacutainer multi-sample needle and screw thread
into a Vacutainer needle holder.
 Disinfect site with a 70% Isopropyl alcohol swab.
 Leave for 30 seconds for the alcohol to evaporate and during this period assemble the
blood tubes required.
 Remove the cover from the multi-sample needle and discard into a clinical waste bin.
 Keeping the needle holder and attached multi-sample needle in one hand use the thumb
on the other hand to press on the vein just above the chosen entry point and pull the skin
back slightly towards you to hold the vein firmly and stretch the skin over the chosen
site.
 With the needle holder and multi-sample needle almost parallel to the patient’s arm and
the needle bevel uppermost, gently push the needle into the chosen venepuncture site.
 Once in the vein hold the needle-holder steady and gently push the cap of the
appropriate blood sample tube onto the covered sample needle at the base of the inside
of the needle-holder.
 Blood should enter the sample tube and fill to the appropriate level indicated.
 Remove the sample tube from the sample needle when full and attach another sample
tube in required.
 Blood samples must be gently mixed at the earliest opportunity to ensure
anticoagulation effectiveness
 As the last blood sample tube is filling slacken the tourniquet by pressing down on the
release clip that is on the side away from the arm.
 Withdraw the needle from the vein and quickly apply a clean pad of cotton wool.
 Ask the patient to keep pressure on the cotton wool to stop further bleeding.
 Discard the needle and holder into a Sharps bin.
 Gently mix the sample tube(s).
 If the patient is unable to maintain sufficient pressure on the venepuncture site apply
this pressure for them.
 Remove the tourniquet from the patient’s arm.
 When bleeding from the venepuncture site has stopped apply Micro- pore tape tightly
over the cotton wool.

17. 3.1.3 PHLEBOTOMY DIAGRAMS

18. 3.2 HEMATOLOGY/IMMUNOHEMATOLGY SECTION
In hematology section, the analysis is carried out using the whole blood sample of
patient for diagnosis of hematological diseases and abnormalities. Blood samples are
collected in EDTA bottle for analysis.
Immunohematology Section Also known as the blood bank performs tests to provide
blood and blood products to patients for transfusion purposes. The blood bank
technologist relies on the phlebotomist to perform identification of the patient without
error, since patients will die if given the wrong blood type diagnosis. The analyses
carried out in these sections include: Packed cell volume, Full blood count, Erythrocyte
sedimentation rate (ESR), Blood film for microfilaria and ABO/D (Rh) typing, Antigen
typing, Blood Genotype, Blood grouping, Cross matching, respectively.
3.2.1 MATERIALS USED IN HAEMATOLOGY/IMMUNOHEMATOLOGY SECTION
Pipette, Hematocrit centrifuge machine for PCV, Hematocrit centrifuge reader for PCV,
Micro Hematocrit analyzer for Full Blood Count, Macro, Microscope, Microscopy slide,
Electrophoresis machine, Cover slip, Bunsen burner, Plasticine, Sterile capillary tube,
Wash bottle, Westergren tubes, Stop watch, Test tubes, Refrigerator, Racks, Various
disposable waste bins, Tiles, Scissors, Ethylene diamine tetra acetic acid (EDTA),
Leishman stain, Normal saline, Water, Antisera for blood group, Buffer solution, Oil
immersion.
3.2.2 BLOOD GENOTYPE (ALKALINE-ACETATE ELECTROPHORESIS)
 Introduction: Hemoglobin electrophoresis often known as Blood Genotype is used to
separate and identify hemoglobin by their migration within an electric field. Blood
variants separate at different rates due to differences in their surface electric charge as
determined by their amino acid structure.
 Aim: To determine the blood genotype of a patient
 Equipment/Apparatus: Tris-EDTA-borate buffer, Electrophoresis machine, Pasteur
pipette, Cellulose acetate paper, gloves, distilled water, Whole blood sample stored in
an EDTA container, Applicator stick, power supply.
 Procedure: Pour 100ml of the Tris-EDTA-borate buffer into the each of the outer
sections zip-zone of the electrophoresis chamber. Wet 2 wicks in the buffer and drape
one over each other support bridge, ensuring each makes contact with the buffer and
there are no bubbles under the wicks. Cover the chamber to prevent evaporation.
Haemolyze the patient’s blood sample accordingly in a well plate and using an
applicator stick apply the samples on the cellulose-acetate paper in small straight lines.
Immediately place the cellulose-acetate paper in the electrophoresis chamber. Connect
the chamber to the zip zone power supply and electrophorese the plate for 25 mins or
shorter at a volt of 350v.
 Result: Hemoglobin H has the highest rate of migration, followed by Hemoglobin A,
then Hemoglobin F, Hemoglobin S and D have similar mobility while Hemoglobin E
and C also have similar mobility.

19.  Conclusion:
AC AS
AA AC
AS SC

20. 3.2.3 PACKED CELL VOLUME (PCV) TEST
 Introduction: The packed cell volume is the volume occupied by the packed red cell
after a volume of anti-coagulated venous blood is fully centrifuged into plasma and red
blood cell. The volume of packed cell is expressed as a percentage of the original
volume of the blood.
 Aim: To estimate the relative mass of red blood cells present in a blood sample in
percentage volume.
 Equipment/Materials: Bunsen burner, Micro hematocrit centrifuge, , whole blood in
an antico Plain non-heparinized capillary tubes (75mm long, with a bore of 1.55-
0.085mm) made from soda lime glass tubes as recommended for blood in EDTA ),
Micro hematocrit reader, an absorbent cotton wool.

21. Procedures
.
 Result: Adult: Normal range for male 37-50%
Normal range for female 35-45%
Children: Normal Range 29-41%
 Conclusion:
Factors affecting the accuracy of PCV are; Unsteady power supply, Poor blood sample
collection, Parallax error while reading the result on the hematocrit reader, Incorrect
blood to anticoagulant ratio, Over spinning of the blood in the centrifuge, Lysing of the
blood by flame or delay in spinning
Bio- medical significance: Low PCV value indicate shortage of blood
3.2. 4 BLOOD GROUPING TEST
 Introduction: A person’s blood group depends on the A,B, or O gene (located on the
chromosome 9) inherited from each parents. ABO blood grouping can be performed
using the principle of antigen-antibody reaction. A person who is blood group A has
anti-body B in their serum. A person who is blood group B, has anti-body B in their
serum. A person with AB blood group has neither anti-body A nor B, in their serum.
Someone who’s O has both anti-body A and B in their serum.
 Aim: To determine the bool type and Rhesus factor of a patient’s blood
 Equipment’s/Materials: Clean free grease tile, Pasteur pipette, Whole blood sample in
an EDTA bottle, applicator stick, test tube rack, clean white tile, cotton wool, applicator
stick, gloves.
 Reagents: Anti-A (BLUE), Anti-B (YELLOW), Anti-D sera (COLOURLESS), Buffer
for balancing, normal saline (GREEN)

22. Procedure: Place 3 drops of patients’ whole blood (usually stored in a green-cap EDTA
container to avoid coagulation) on three different spots in a clean grease-free white tile. Add
a drop of ANTI-A, ANTI-B and D reagents simultaneously on these spots. Mix the contents
of each division using a clean piece of applicator stick for each. Tilting gently the tile from
side to side, look for agglutination after 2mins, then record your results.
N/B: Although agglutination Is usually seen well within 2 mins, the full time must be
allowed. In order to detect a weak reaction.
 Result:

23.  Conclusion: The result was observed according to the agglutination that occurred in
each spots on the tile. Anti D determines the present of the rhesus ‘D’ factor in blood
group.
Factors that affect blood grouping are; wrong labeling of spot and confusion of anti-
sera with spots, Contamination of test card or tiles with detergents, Expired anti-sera,
not haemolyzing the blood sample with the Anti-gen properly.
Bio-medical significance; Blood transfusion, Blood compatibility, Antenatal
screening#
3.3 SEROLOGY SECTION
Tests done in this department are designed to detect the body's response to the presence of
bacterial, viral, fungal, parasitic and other conditions which stimulate detectable antigen-
antibody reactions in a test system to aid in the diagnosis of the patient. Most tests
performed in this section are carried out under the principles of Immunoassay, some of
them are; Cold agglutinins (CAG) - specimen must be kept at room temperature, Widal,
Asotitre.
3.3.1 FOR HEPATITIS USING HBs.Ag TEST STRIP
 Introduction: HBsAG is a rapid immune-chromatographic test for the qualitative
detection of Hepatitis B surface Antigen in human serum/plasma, it can be used for
prenatal or transfusion screening, and during acute infection or chronic carriage of the
Hepatitis B virus. Early detection of infection is essential for rapid initiation of adequate
treatment.
VDRL test is a screening test for syphilis. It measures substances called antibodies that
body may produce if it comes in contact with the causative agent of syphilis, which is
called Treponemapallidium
 Aim: To determine the presence or absence of hepatitis and syphilis in the body
system.
 Materials: HBsAG Test strips, VDRL test strip EDTA bottle, Centrifuge, clean test
tube
 Specimen: Serum.
 Procedure: The patient blood sample was collected into a plain bottle through
venipuncture. The blood sample was spun in a centrifuge for 5 minutes, after spinning
the serum was separated carefully into a clean test tube by the use of Pasteur pipette and
then test strip was immersed vertically into the serum for 10 minutes. The observation
was taken after 10mins.
 Result: Appearance of a line at the Control region and another at the Test indicates
positive result, while an appearance of a line at the Control region only, indicates
negative result. When there is no appearance of any line, means the test in invalid and
as to be redone using new kits

24. 3.3.2 WIDAL TEST
 Introduction: Typhoid fever is an infectious disease caused by the Salmonellatyphi, it
is diagnosed by Widal test which employs an antigen-antibody reaction to screen for the
presence of Salmonellatyphiand paratyphiantibodies in the sample serum.. the organism
is transmitted by water or food contaminated by faeces of typhoid victims or carriers,
that is a person who harbor it without showing signs and symptoms.
 Aim: To investigate the presence of Salmonellatyphi and paratyphi in the serum of
patient
 Materials: Test card/white tiles, Pasteur pipette, centrifuge, antigen kit and stop watch
 Procedure: 3-5ml of blood was collected from the patient through venipuncture into a
plain bottle and the blood was spun at 3000rev per min for 5minutes so as to separate
out the plasma. A dropper was used to carefully drawn the antigen kits and a drop was
placed on each of the test card in pairs of four spots labeled O, OA, OB, OC and H, HA,
HB, HC and a drop of serum was carefully added into the antigen respectively with the
aid of Pasteur pipette and mixed together with the aid of an applicator stick the test card
was rocked gently, the rate of reaction and agglutination was observed at an interval of
30sec, 1min, 2mins, and 5mins
 Specimen: Serum
 Result
Reactive: visible agglutination on spot H and others indicate the present of Salmonella
antibodies
Non-reactive: no visible agglutination indicates absence of Salmonella antibodies
Widal test: Positive
Highly reactive……………………………….1:320 (agglutination reaction before 60
seconds)
Very reactive……………………………….1:160(agglutination reaction before 120
seconds)
Reactive ………………………………………1:80(agglutination reaction before 180
seconds)
Widal test: Negative
Non-significant…………………………………1:40
Non-significant………………………………….1:20
Not reactive…………………………………….Nil

25. 3.3.3 HORMONE PROFILING [MALE+FEMALE] (FSH, LH, EST, PROG,
TESTESTERONE, PRL)
Introduction: Hormone profile test helps you to understand the make-up of your hormones,
as well as detecting imbalances and to reveal important information about the rate of
production of these hormones in a patient’s body e.g.: Fertility level, Steroids, and Thyroids
Fertility hormones include: Follicle Stimulating Hormone [Male & Female], Luteinizing
Hormone [Male & Female], and Prolactin. While Steroids include: Testosterone [Male],
Progesterone [Female], and Estradiol [Male & Female].
Aim: To investigate and detect hormonal imbalances in the Male and Female body by using
blood serum.
Materials: A sample of a patients’ lysed serum, Micro-well, Micro-well washer, Microplate,
Micro-well reader, Accubind Eliza reagent Kits, pipette 100ul, Eliza reader, stop watch or
timer.
Specimen: Serum
Procedure: Pipette 100ul of sample into the micro-well and leave for 5mins before pipetting
100ul of reagents [either one of FSH, PRL, PRG, LH, E2, or T4] leave for 1 hr. Before
washing with a micro-well washer, the mixture of the samples are pipetted out and then
washed at a pipetted value of 1000ul for 3 times. After washing you decant, and it is advised
to avoid touching the bottom of the micro-well when decanting as the Micro-well reader
reads vertically. The sample is now left for few minutes before it is taken to the other section
for reading with the micro-well reader.
Male:
HORMONES…………………..MNIMUM RANGES ……………MAXIMUM RANGE
FSH 1.0ng/ml 14.0ng/ml
E2 19
LH 0.7ng/ml 7.4ng/ml
PRL 1.8ng/ml 17.0ng/ml
TSH 0.28ng/ml 0.53ng/ml
Female:
HORMONES…………………MINIMUM RANGES……………MAXIMUM RANGE
FSH
[Follicular phase] 3.0 ng/ml 12.0ng/ml

26. [Midcycle phase] 8.0ng/ml 22.ong/ml
E2
Follicular phase: 48ng/ml 9.175mlu
Luteal Phase: 103ng/ml 196ng/ml
Periovulatory phase: 209ng/ml 281ng/ml
Treated Menopause: 122ng/ml 289ng/ml
Untreated Menopause: 7.3ng/ml ND20
LH
Follicular Phase: 0.5ng/ml 10.5ng/ml
Midcycle phase: 18.4ng/ml 61.2ng/ml
Luteal Phase: 0.5ng/ml 10.5ng/ml
PRL
Adult women 1.2ng/ml 19.5ng/ml
Postmenopausal 1.5ng/ml 18.5ng/ml
3.3.5 ANTI-STREPTOLYSIN O TITRE TEST (ASOTITRE)
 Introduction: This is for the diagnosis of the measurement of anti-bodies fighting
against streptolysin a substance produced by a group of A streptococcus bacteria. This
streptococcus bacteria’s cause different forms of disease ranging from sore-throat,
cold, catarrh, impetigo etc. Some of its symptoms are also Yellow Fever.
 Aim: To investigate the level of steptolysin in a patient’s blood by using anti-
streptolysin reagents
 Materials: Positive/Negative Anti-serums kit, Asotitre slides with a positive/negative
control, Applicator stick, Pasteur pipette, and result booklet.
 Specimen: Serum
 Procedure: The blood sample collected in the plain bottle was centrifuged for 15mins
till it was lysed. The serum was collected using a Pasteur pipette, and 3 drops of the
sample was placed on the Asotitre slide for both the positive and Negative control
circles. A drop of ASO latex antigen was added and mixed with the sample using a
disposable applicator stick, after that the slides were rocked back and for 2 mins and
examined for agglutination.
 Result: Agglutination is a positive result and indicates the presence of detectable Anti-
streptolysin-o antibody is the serum of the patient. No agglutination is a negative test
result and indicates the absence of detectable anti streptolysin-o antibody in the serum

27. Diagram depicting and Asotitre kit and showing the +ve and –ve glass slides
3.4 CLINICAL CHEMISTRY SECTION
This section deals with chemical analysis of serum or plasma in which many diseases of
the major organs systems can be diagnosed such as heart attacks, hepatitis, renal failure,
diabetes, Liver function, etc. Blood sample samples may collected into the Serum
Separator Tube or Lithium Heparin. Test performed in this department are:
 Liver function Which include four parameters Include: AST, ALT, ALP, and Bilirubin.
3.4.1 LIVER FUNCTION TEST
 Introduction: This is a group of blood tests that detects inflammation and damage to
the liver and also checks how well the liver works. Liver function test includes ALT
(Alanine amino transferase)
AST (Aspartate Aminotransferase) others are PT, INR, albumin, bilirubin
ALP (Alkaline phosphatase)
 Materials: Precision Kit, Biochemistry Analyzer, Test tubes, Micro pipette 100ml,
ALT reagent, AST reagent, ALP reagent, Bilirubin Kit, Timer
 Specimen: Serum
 Procedure: Blood is collected using an EDTA container and left to be centrifuged
until the patients’ serum is utilized. After that the 10U/L sample is added with
(1000u/l for both ALT&AST reagents). Also Alp has two reagents of which use
500u/l each into the sample. The sample to left to examined using the Biochemistry
Analyzer for about 20-25Oc for 5mins.
 Result:

28. 3 .5 .1 MICROBIOLOGY SECTION/PARASITOLOGY SECTION
Microbiology involves the study of microbes. Although, microorganisms are generally
beneficial and essential to life some are, however, pathogenic and cause infectious
diseases. The diagnostic microbiology laboratory is engaged in the identification of
infectious agents, WHILE the Parasitology bench deals with the relationship of some of
these parasites with their Host and help to effective stop their parasitic load on their host.
These infectious agents are broadly classified as viruses, bacteria, mycotic agents and
parasites (Protozoans and Helminthes) for both microbiology and Parasitology
respectively. Also this section analyses body fluids and tissues for the presence of
pathogenic microorganisms primarily by means of culture and sensitivity (C&S). Results
of the C&S tell the physician the type of organisms present as well as the particular
antibiotic that would be most effective for treatment.
BASIC RULES FOR WORKING IN THE MICROBIOLOGY LABORATORY,
 While working in the laboratory, it is important that you must adhere to the following
basic rules;
 Be methodical and orderly in habits; keep the work area clean especially before leaving
the laboratory and disinfectant it thoroughly at the end of day.
 Wash hands frequently with soap and water
 Before leaving the laboratory, place the discarded glassware into the designated place.
 Cultures are kept under incubation and should be inspected in the morning and findings
must be carefully.
 Send the laboratory reports promptly. In case of emergency a special report is
dispatched or communicated by telephone.
3.5.2 GENERAL REMARKS
 All specimens for culture must be collected prior to the therapy. If the patient is on
antibiotic, inform the microbiologist so that he/she can take measures.
 Collect the specimen in adequate amount from the infectious site. This usually
instructed by the physician.
 Always use the sterile bottle to transporting the specimen.
 All specimen must be accompanied by a request slip with complete information, h on
the patient name, age, sex, hospital number, source of specimen and clinical information
is very important in order to choose the appropriate medium for the culture.
 Enter the details in the laboratory register and performed direct examination of the
specimen before choosing the media for the culture.
 All containers used for holding microbiological specimens must be sterilized before
used. Such as test tube, culture tube with and without cap, and plates, container to
collect sputum specimen, blood specimen for microbial culture, penicillin bottles for
collection of spinal fluid and other specimen container (universal bottle) for collection
of urine specimen and stool specimen.

29. 3.5.2 MATERIALS/APPARATUS USED IN MICROBIOLOGY/PARSITOLOGY
SECTION
Materials include: Inoculating loop, Bunsen burner, Incubator, Weighing balance, Spatula,
Microscopy slide, Cover slip, Staining rack, Medium plates, Sensitivity disc, Forceps,
Cotton wool swab, Petri-dish, Flat-bottom flask, and Beakers.
3.5.3 INTRODUCTION LABORATORY GROWTH MEDIA
These are classified into six types: (1) Basal media, (2) Enriched media, (3) Selective
media, (4) Indicator media, (5) Transport media, and (6) Storage media.
1. Basal Media. Basal media are those that may be used for growth (culture) of bacteria
that do not need enrichment of the media. Examples: Nutrient broth, nutrient agar and
peptone water. Staphylococcus and Enterobacteriaceae grow in these media.
2. Enriched Media. The media are enriched usually by adding blood, serum or egg.
Examples: Enriched media are blood agar and Lowenstein-Jensen media. Streptococci
grow in blood agar media.
3. Selective Media. These media favour the growth of a particular bacterium by inhibiting
the growth of undesired bacteria and allowing growth of desirable bacteria. Examples:
MacConkey agar, Lowenstein-Jensen media, tellurite media (Tellurite inhibits the growth
of most of the throat organisms except diphtheria bacilli). Antibiotic may be added to a
medium for inhibition.
4. Indicator (Differential) Media. An indicator is included in the medium. A particular
organism causes change in the indicator, e.g. blood, neutral red, tellurite. Examples: Blood
agar and MacConkey agar are indicator media.
5. Transport Media. These media are used when specie-men cannot be cultured soon
after collection. Examples: Cary-Blair medium, Amie’s medium, Stuart medium.

30. 6. Storage Media. Media used for storing the bacteria for a long period of time.
Examples: Egg saline medium, chalk cooked meat broth. The survival and growth of
microorganisms depend on available and a favorable condition environment. Culture
media are nutrient solutions used in laboratories to grow microorganisms. The method for
the preparation of basic microbiology media is given below. Sterilization is at 121c for
15minutes.pH values are 7.0 unless stated otherwise.
3.5.4 COMMON MEDIA IN ROUTINE USE
1. CLED Agar (Cysteine Lactose Electrolyte Deficient): is a non-selective differential
plating medium for the growth and enumeration of urinary tract microorganism.
Preparation: 36.0g of medium is suspended in one liter of distilled water, slowly heated
and frequently stirred. Boiled for a minute and sterilized at 121O
C for 15minutes and
poured into Petri dishes. Plates were inverted when solidified for storage purposes and to
avoid moisture
2. MacConkey Agar. Most commonly used for enterobacteriaceae. It contains agar, peptone,
sodium chloride, bile salt, lactose and neutral red. It is a selective and indicator medium:
a) Selective: as bile salt does not inhibit the growth of enterobacterceae but inhibits
growth of many other bacteria.
b) Indicator: medium as the colonies of bacteria that ferment lactose take a pink colour
due to production of acid. Acid turns the indicator neutral red to pink. These bacteria
are called 'lactose fermenter', e.g. Escherichia coll. Colourless colony indicates that
lactose is not fermented, i.e. the bacterium is non-lactose fermenter, e.g. Salmonella.
Shigella, Vibrio.
Preparation: 50g of the Agar was suspended and measured into one liter of purified
water and mixed thoroughly and was heated with frequent agitation, then boiled for
one minute to completely dissolve the powder. The Agar was autoclaved at 121O
C for
15minutes.
3. Chocolate Agar or Heated Blood agar. Prepared by heating blood agar. It is used for
culture of pneumococcus, gonococcus, meningococcus and Haemophilus. Heating the
blood inactivates inhibitor of growths.
Preparation: 2litres of distilled water was added to 144g of agar powder. Autoclaving
was done at 121O
C for 15minutes and cooled till 45O
C then 5% of defibrinated blood was
added. Heated slowly and evenly to 65O
C, cooled till 45O
C and poured into plates
4. Blood Agar. Most commonly used medium. 5-10% defibrinated sheep blood is added to
melted agar at 45-50°C. Blood acts as an enrichment material and also as an indicator.
Certain bacteria when grown in blood agar produce haemolysis around their colonies.
Certain bacteria produce no haemolysis. Types of changes:
(a) Beta () haemolysis. The colony is surrounded by a clear zone of complete haemolysis,
e.g. Streptococcus pyogenes is a beta haemolytic Streptococci.
(b)Alpha () haemolysis. The colony is surrounded by a zone of greenish discolouration
due to formation of biliverdin, e.g. Viridian’s streptococci, and
(c) Gamma () haemolysis, or, No haemolysis. There is no change in the medium
surrounding the colony,

31. 5. Nutrient Broth. 500 g meat, e.g. ox heart is minced and mixed with 1 litre water. 10g
peptone and 5 g sodium chloride are added, pH is adjusted to 7.3. Uses: (1) As a basal
media for the preparation of other media, (2) To study soluble products of bacteria.
3.5.5 DISPOSAL
Once the Petri dishes have been taped shut, they should not be opened again. All
microorganisms grown during the experiment should be killed before discarding. The best
way to dispose of bacterial cultures is to pressure sterilize them in a heat stable biohazard
bag. If autoclaves or pressure cookers are not available or large enough to make this
convenient, an alternative is to bleach the plates. Saturate the plates with a 20% or "1 in 5"
household bleach solution (in other words, 1part bleach and 4 parts water). Let them sit
and soak overnight in the bleach solution before disposing of them. Please note that the
bleach solution is corrosive and needs to be thoroughly removed afterwards. In addition,
the plates can be incinerated if access to an incinerator is available.
3.5.6 PRECAUTIONS TAKEN WHEN PREPARING MEDIUM
Do not talk when pouring medium on plates and when culturing the sample on plates to
avoid contaminants as a result of unwanted bacterial or enzymes through saliva.
The degree at which we incubate any cultured sample is always at 37c to avoid the death
of the microorganism.
3.5.7 GRAM STAINING
 Introduction: In this section, the staining of bacteria as a means for identification is
done. Bacteria are being identified as Gram-Negative or Positive on the basis of their
cell wall thickness after staining. Gram positive bacteria hold the dye and appear purple
while Gram-Negative bacteria release the dye and appear red.
 Aim: To identify the gram-negative and the gram positive bacteria.
 Apparatus: Stop watch, Normal saline, Clean grease free microscopic slide, Gentian
violet, Lugol’s iodine, 95% ethyl alcohol, Neutral red, Microscope, Sterilized
inoculating loop.
 Procedure: The organism was isolated and smeared using the sterilized inoculating
loop in a drop of normal saline on a clean grease free microscopic slide. It was left to
Air-dry.
The smear was placed on a staining rack, and was flooded with Gentian violet solution
for 30seconds. It was rinsed with water. The smear was again flooded with Lugol’s
iodine for 30 seconds. It was rinsed with water and the smear was decolorized with 95%
ethyl alcohol for 30 seconds, It was rinsed with water again.
The decolorized smear was counter-stained by flooding with neutral red for 30 second
and was rinsed with water. The back of the slide was cleaned with cotton wool and
allowed to Air-dry. The slide was mounted on the microscope after air-drying and was
examined under ×100 immersion objectives. The result was recorded.
 Sensitivity Test (Result): If the bacteria are gram positive, a positive sensitive disc is
used while a negative sensitive disc is used for gram negative bacteria-A pure colony
was sub-cultured on a Nutrient medium and sensitive disc was picked with the aid of a

32.  sterile forceps, and placed on the medium, then the plate was incubated for 24 hours at
37O
C.The plate was read after 24hours of incubation to observe zone of inhibition and
resistance. Sensitivity test is also done for pathogens that grow on the media by taking a
colony of the organism, streak on the sensitivity agar and add antibiotics discs and
incubate for 24hours at 37O
C.
3.5.8 PROCEDURE FOR AUTOCLAVING
The inner element was filled with water and allowed to cover the surface of the element,
The medium were arranged in the sample bucket and the machine was covered and
screwed tightly by holding the screw opposite each other and the wing nut was screwed
tightly.
The switch on button was on and also the outlet valve was opened down so as to increase
the temperature of the steam on the workload.
It was sterilized at 121⁰C for 15minutes. The safety valve was closed at 120⁰C and at
121⁰C, button was switched off. The medium were allowed to cool for 47⁰C before
pouring in the Petri dish
Principle of Autoclaving:
The principle behind these sterilization methods is based on the temperature and the type
of autoclaving operation performed. Autoclaving operation at 121⁰C is referred to as
culture media autoclave.
3.6 MICROSCOPY, CULTURE AND SENSITIVITY TESTS
Microscopy involves the examination of specimen under the microscope, Culture refers to
the microorganisms that grow on the culture medium after inoculation and incubation
while sensitivity tests determines the antibiotics which will be administered to the patients.

33. The cultured plates are incubated for 24hours at 37o
C to facilitate the growth of the
organism and chocolate plates were incubated in a candle jar to facilitate the growth of
both aerobic and anaerobic microbes while other plates were incubated aerobically.
3.6.1 STOOL ANALYSIS/PARASITES
 Introduction: Stool analysis involves the collection and analysis of faecal matter to
diagnose the presence or absence of a medical condition. During the outbreak of cholera
or diarrhea, food microbiologist collect stool sample into sterile universal bottle from
victims for fecal examination in the laboratory.
 Aim: To determine cysts and ova in the stool of a patient.
 Materials/Apparatus: Test tubes, Clean microscopic glass slide, drop of saline,
binocular microscope, swab stick, coverslip, loop, physiological Saline, TCBS agar,
MIU agar, Selenite F Broth,
 Procedures:
FOR SALMONELLA, SHIGELLA AND E.COLI:
After the reception has the labelled appropriately the Name, Age/sex, Examination
required, and the Hospital where the patients’ stool was brought:
1. Incubate a well dried DCA plate with a heavy inoculum (3 loopfuls of
unformed/watery or emulsified ½g of formed stool in 2mls of normal saline),
preferably from mucoid or bloody portion (if present). Also inoculate a loopfuls of the
stool onto a well dried MacConkey agar, streaking in way as to obtain discrete colonies
after incubation. Inoculate selenite F broth of about 4mls with about 1g of stool.
2. Incubate at 37°C for 18-24hrs.
3. Subculture the selenite F broth onto a dry DCA and MacConkey agar plate and
incubate again at 37°Cfor 18-24hrs.
4. Read the primary plates for NIF and circle with grease pencil and select similar
colonies for sugars and other biochemical tests.
5. With a straight wire, pick each selected colony and inoculate into peptone water, urea
broth and Simon's citrate medium. Incubate for 4hrs at 37°C and using peptone water
inoculate other sugars (glucose, lactose, mannitol, sucrose and xylose. Alternatively,
inoculate KIA or TSIA, MIU agar. Incubate at 37°C for 18-24hrs.
6. From the primary MacConkey agar plates, touch 5 different discrete colonies of
suspected E. coli (if any) and subculture onto another MacConkey agar, incubate at
37°C for 18-24hrs. Use this for EPEC typing using EPEC antisera.
7. Perform step 5 with selenite F broth subculture after 18-24hrs incubation.
8. On the 3 and 4* days, read the sugar reactions for all the isolates. Perform other
biochemical tests e.g. indole and oxidase.
9. Type for Shigella or Salmonella using the appropriate antisera.
Results: The results were viewed in two ways which are microscopic and macroscopic;
1. Macroscopic-----------------Hard, dark ,brown stool formed with no mucus or blood
seen.
2. Microscopic-----------------Present of Ascaris lumbricoides.

34. 3.6.2 URINE MICROSCOPY CULTURE AND SENSITIVITY (M/C/S)
Urine for microscopy culture and sensitivity is an array of tests performed on urine
samples to examine the presence or absence of cells such as; epithelial cells, pus cells, red
blood cells, yeast cells, crystals, and bacteria. It is one of the most common methods of
medical diagnosis.
 Aim: To identify parasites and Bacteria cell in the urine of individual.
 Materials: Microscope, Sterile container [for collecting patient’s urine] microscopy
slide, centrifuge, inoculating loop, urine sample, media plates; MacConkey agar and
CLED agar, Blood Agar, Nutrient agar, Gram staining reagents, Sensitivity disc,
Incubator, Bunsen burner.
 Specimen: Early morning Urine [EMU], Mid-stream Urine [MSU]
 Procedures: Urine collected should be processed within 1 hr. of collection or
refrigerated at 4°C
 Microscopy: DAY 1
Note the appearance e.g.: Cloudy, Blood-stained, Straw-colored
Macroscopy: DAY 1
Examine uncentrifuged and unstained urine samples and note the number of WBC per
field, number of RBC, epithelial cells, bacteria, casts, crystals etc.
Culture: We used a standard inoculation loop [0.01ml], was streaked onto a blood Agar,
MacConkey or CLED media, and incubated at 35-37°C
DAY 2: Examine plates for growth. Note the number and the type of growth (significant
bacteriuria>10^3cells/ml) and gram stain
Gram Staining: Primary and secondary gram staining was done on the smear of a colony
of the bacteria. This is to identify if the bacteria is Gram positive or Gram negative.
Do Biochemical tests and antimicrobial susceptibility testing at 35-37°C overnight.
fig 2.12 An antimicrobial susceptibility testing disc showing zones of inhibition.
3.6.3 FUNGAL/MYCOLOGY CULTURE

35.  Materials: KOH (20% and 40%), 70% ethanol, lacto phenol cotton blue, scalpel
blade, tweezers, scissors, glass slides, cover slips, Sabouraud Dextrose agar or Malt
extract agar.
Procedure: SAMPLE COLLECTION
. Cleanse the affected area with 70% (v/v) ethanol.
 Collect skin scales, crusts, pieces of nails or hairs on a clean piece of paper as
follows:
 1. Skin scales: Collect by scraping the surface of the margin of the lesion using a
sterile scalpel blade.
 2. Crusts: Collect by removing part of the crust nearest to healthy skin using sterile
scissor and tweezers.
 3. Nail pieces: Collect by taking snipping of the infected part of the nail using
sterile scissors.
 4. Hairs: Collect by removing dull, broken hairs from the margin of the lesion using
sterile tweezers. Fold the paper to form a flat packet after collecting and label
appropriately. Submit to the lab.
 Processing the sample: КОН
Place a drop potassium hydroxide solution (20% w/v for skin and 40% w/v for hair and
nail clippings). Transfer the specimen (small pieces) to the drop of KOH, and cover
sup. Place the slide in a Petri dish or other container with a lid and containing a piece
of damp filter paper or cotton wool to prevent the preparation from drying out. Leave
for some time for the specimen to clear. As soon as the specimen has cleared, examine
microscopically using 1Ux and 40x objective with the condenser iris sufficiently closed
to give good contrast.
 Note: clearing can be hastened by gently passing the preparation over flame or
adding 40% Dimethyl sulfa-oxide solution (DMSO).
Look for branching hyphae, chains of angular or rounded arthrospores or a mixture
of both.
 Culture: Using a sterile blade or scissors (flame-sterilized) cut the specimen into
pieces as small as possible. Using sterile tweezers, inoculate the small pieces (a few
millimeters apart) on the surface of malt extract agar or Sabouraud Dextrose agar.
Incubate at room temperature (25-30 C) for up to 3weeks, examining every few
days for growth.
3.6.4 URO-GENITAL, URETHRAL AND ENDOCERVICAL SWAB.
 Test Overview: This is use to detect the causative organisms of female reproductive
system infections and their sensitivity to antibiotics.
 Procedure: FOR VAGINAL SWAB:

36. The patient should lie on her back, knee drawn up and the thighs spread apart to reveal the
vagina. Remove the swab from the plastic wrapper without touching the cotton tip or
laying it down. Insert the swab into the vagina about inches deep. Gently rotate clockwise
for 10 to 30 seconds. Withdraw the swab without touching the skin. Replace the swab in
the plastic wrapper or place in a transport medium, label correctly and send to the
microbiology laboratory.
MICROSCOPY:
Wet Preparation
Make a wet prep by emulsifying the swab in few drops of normal saline
Cover with a cover slip and examine using ×40 objectives.
Report the number of pus cells and reds per high power field. Also indicate the presence of
Yeast cells, epithelial cells, Trichomonas vaginalis etc.
Gram Stain Procedure
A direct smear for Gram staining may be performed as soon as the swab specimen is
collected from the urethra, cervix, vagina or rectum. The swab should be rolled gently onto
the slide to preserve cellular morphology and over an area less than 1 cm?
Allow the smear to dry.
Heat-fix over a Bunsen burner; ensure the heat is not so much as to irritate the back of the
hand.
Stain with crystal violet stain for 1 minute
Wash off in gentle running tap water
Flood with Lugol's iodine for 1 minute and wash in running tap water.
Decolorize with acetone alcohol for a few seconds
Wash in tap water.
Counter stain with satranin red.
Wash and allow the stain to dry before examining it using x100 objectives
CULTURE
DAY 1
Inoculate the sample on MacConkey, Chocolate and Blood agar media and incubated over-
night at 35-37 C.
DAY 2
Examine the plates for growth. Note the number and type of growth (Significant bacteriuria
>10° cells/ml).
Gram stain the colonies
Carry out biochemical tests.
Set up antimicrobial susceptibility testing. Incubate at 35-37°C overnight.
DAY 3
Note the zone of inhibition of the different antibiotics used.
Report the degree of susceptibility using NCCLS chart and dispatch your results.
3.6.5 MALARIA PARASITE TEST
LABORATORY DIAGNOSIS OF MALARIA PARASITE
Malaria is a mosquito borne disease transmitted by the bite of a female Anopheles
mosquitoes, malaria causes symptoms such as headache, fatigue, fever, vomiting etc. The

37. bite from a female anopheles mosquito introduces the parasite from the mosquito saliva
into ones blood stream.
AIM: To confirm if Trophozoites of Plasmodium falciparium is present in the Blood sample
of a patient
MATERIALS NEEDED: EDTA bottle containing the blood sample, Malaria staining
reagent, field stain A and B, Slide and Slide racks, and Immersion oil
PRECAUTION
1. Make sure that the bottle is an EDTA bottle so as to avoid coagulation.
2. There should be no spillage of blood on the bottle, The specimen bottle should be
labeled properly.
PROCEDURES
1. Disinfect the working bench with alcohol.
2. Label the slides with the patient’s number.
3. Using a completely clean grease free slides, spread a large drop of the blood to
make a thick smear of the blood on one slide, making it possible to see news print
through the film.
4. On the second slide, a little quantity of blood was dropped on the Centre of the
slide, and the thin film was spread from an angle using a smooth edge slide spreader
so as to make a smooth tail end.
5. Allow the side to air-dry by placing it in a horizontal position away from insect and
dust.
6. After drying the slides, the thick film was dipped into field stain A for 5seconds
and excess stain drained
7. The slide was then rinsed gently in a clean water and drained
8. Dip the thick film slide into field stain B for 3 seconds, drain gently and wipe the
back of the slide clean and allow to dry.
9. For the thin film the reverse was the case, it can be fixed with methanol before
staining , the thin film was first dipped in field stain B for 3 seconds, rinsed off in a
clean water and field stain A for 5 seconds. The thin film slide was rinsed and
allowed to dry properly.
10. Immersion oil was added and viewed with X100 objective lens.
RESULT
Less than 10 Trophozoites of Plasmodium falciparium with a cytoplasm and chromatin dot
was observed.

38. CHAPTER FOUR
4.0 SUMMARY, CHALLENGES ENCOUNTERED, AND CONCLUSION
RECOMMENDATION
4.1 SUMMARY OF ATTACHMENT ACTIVITIES
During my period at the De-Family Health Diagnostic & Research Centre, Imo
State as a SIWES student, I aided in the cataloguing some information materials for the
laboratory, I also did some activities at the laboratory reception such as: attending to
patients, confirming and examining their request forms, entering their details into the
computer system and then to the laboratory register, and also running phlebotomy upon
Med. Scientist request.
4.2 CHALLENGES ENCOUNTERED
The main constraint that I had encountered during my 5 months of S.I.W.E.S was
the issue of IT placement. During my search for placement at a Lab some many
Organizations rejected Me especially Laboratory Institutions, the claim was that Most
students are sent out by lecturers to gather information that are being used to secure
their Master’s Degree programs, and because of that the Union of Medical Laboratory
Scientist’s has restricted any course that has nothing to do with Medical Lab. Therefore,
the need to protect their profession has been an issue for most of us doing other courses
in school. It was also quite challenging for me because the Laboratory I happened to
work with was very far from my home and Transportation fare became unbearably
expensive, coupled with the lack of cash influx during the election period. Other
problems I encountered during the training was attending to different people with
different personalities at the reception.
4.3 CONCLUSION
My five months industrial attachment with De-Family Health Diagnostic and
research Centre has been one of the most interesting, productive, instructive and
educative experience in my life. Through this training, I have gained new insight and
more comprehensive understanding about the real industrial working condition and
practice, thereby improving on my theoretical skills that I have gained over the past 3
years.
All these valuable experiences and knowledge that I have gained were not only
acquired through the direct involvement in task but also through other aspects of the
training such as: work observation, supervision, interaction with colleagues,
supervisors, superior and other people related to the field. It also exposed me to some
certain things about medical environment. And from what I have undergone, I am sure
that the industrial training program has achieved its primary objective.
As a result of the program, I am now more confident to build my future career
which I have already started with De-Family Health Diagnostics.
4.4 RECOMMENDATION
To students that are to undergo the training, I recommend that they should take it very
seriously, because it is one of the most important parts of their education which will
help them build a very significant and effective meaning in their career pursuit.