Government College University

Faisalabad

INTERNSHIP REPORT
By
NIMRA AZIZ
BS (Hons.) Physiology
3583

8th Semester
Department of Physiology

“DEPARTMENT OF PATHOLOGY, FAISALABAD

INSTITUTE OF CARDIOLOGY”
 Internship completion certificate

It is certified that the research work in this report, has been carried out and completed by
“NIMRA AZIZ” Registration No. 2012-GCUF-02509 under my supervision during her BS
(Hons.) studies in the subject of “PHYSIOLOGY” as a student of “DEPARTMENT OF
PHYSIOLOGY”, Government College University Faisalabad.

SUPERVISOR SUPERVISOR

DR. MUHAMMAD AMEEN DR. HASEEB ANWAR

Consultant Pathologist Assistant Professor
Head of Pathology Department Head of Physiology Department
Faisalabad Institute of Cardiology Government College University
Faisalabad Faisalabad
 Preface
Internship training program provides a chance to students to make
themselves Familiar with the actual professional environment. Working in the
clinical laboratory reflects almost all the practical work that are learned
during theoretical courses.

This report documents the work done during the internship at Faisalabad
Institute of Cardiology (FIC).This report covers all the activities that have
contributed to achieve a number of stated goals. In the following chapters the
report first will give an overview of the tasks completed during the period of
internship with technical details. Then the results obtained will be discussed
and analyzed.
 Acknowledgement
First of all, I would like to express my deepest gratitude to Almighty Allah for
giving me strength and composure to complete this report within scheduled
time.
I am thankful to the Supervisor DR.MUHAMMAD AMEEN, Consultant
Pathologist and Head of Department of Pathology, Faisalabad Institute of
Cardiology (FIC) for giving me the opportunity to do my internship at his
highly esteemed organization.
I am also grateful to DR. HASEEB ANWAR Head of Department of
Physiology, Government College University Faisalabad who equipped his
students with knowledge and made them able to do internship in such
valuable, esteemed and professional institute.
I am also thankful to the Co-supervisors Dr. Ameer, Dr. Kashif, Dr. Mubashir
And Madam Asma for their valuable guidance, suggestions, advice and
encouragement.
I am extremely grateful to all the people working in the laboratory for their
reception and friendliness and for giving me information and valuable
guidance during the period of internship.
Thanks to them, my time in Faisalabad Institute of Cardiology Laboratory
was an enriching experience and it was real pleasure to work with them.

NIMRA AZIZ
Contents
CHAPTER NO. 1 INTRODUCTION ........................................................................... 1
CHAPTER NO. 2 LAB ETHICS .................................................................................. 3
CHAPTER NO. 3 PHLEBOTOMY AND MAIN RECEPTION ........................................ 5
CHAPTER NO. 4 HEMATOLOGY ............................................................................. 9
CHAPTER NO. 5 CLINICAL BIOCHEMISTRY ........................................................... 14
CHAPTER NO. 6 MICROBIOLOGY ......................................................................... 19
OPERATION THEATER LAB ................................................................................... 31
Learning Outcomes: ............................................................................................ 32
CHAPTER NO. 1 INTRODUCTION
Introduction of Hospital:
Faisalabad Institute of Cardiology, Faisalabad started working on November 13, 2007. This
institute is a tertiary level specialized autonomous institution for cardiac disease. It consists of
202 beds. It is situated near Chenab Club, Faisalabad.
This institution is providing services to cardiac patients of Faisalabad and its adjacent districts

like Sargodha, Toba Tek Singh, Jhang, Chiniot and beyond areas. This is a teaching institute
providing post graduation in Cardiology, Cardiac Surgery and BS in Cardiac Perfusion.

Mission Statement:
Faisalabad Institute of Cardiology strives to play a leading role in providing the Evidence Based
Healthcare services to cardiac patients and to prepare Healthcare Professionals for the challenges
of future through Academic Activities.

Objective Statement:
The objectives of this institute are:
 To provide the best possible cardiac diagnostic and treatment facilities to the people of
Faisalabad and other adjoining areas.
 Improve Cardiac Health care facilities in this region.
 Provide facilities of highly specialized nature.
 Act as a referral center for the central Punjab and adjacent areas.
 Train medical, paramedical personnel and nurses, capacity building in this field.
 Provide research facilities in the field of cardiology and cardiac surgery.
 Provide training in all Cardiac specialties to medical graduate.

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  Attract patients and research workers for advanced and specialized training and research
work.

Facilities:
 It is a 202 bedded hospital.
 It has well equipped operation theaters.
 10 OPD clinics are providing services to 800 to 1000 patients per day.
 Routine cardiac drugs are being provided to needy patients free of cost.
 FIC has two 10 bedded CCUs and 18 bedded emergency ward with bedside cardiac
monitoring, central Oxygen and suction facility. All other equipment required for the
treatment of cardiac emergencies like defibrillators, ventilators and temporary
pacemakers are also available.
 Every test for every cardiac patients round the clock.

Training procedure:
 Supervisor and Co- supervisor is available round the clock.
 Working in every section of lab periodically.
 Practical demonstration and active working assistance during analytical working in all
modules.
 Standard sample collection according to SOPs.
 Blood gases estimation.
 Sample fitness/ labeling for analysis.

Department of Pathology:
The FIC provides a swift, accurate and dependable diagnostic laboratory services that are highly
affordable and competitive. The pathology laboratory is under the supervision of Dr. Muhammad
Ameen.
 About 40,000 to 42,000 tests are performed in this laboratory per month.
 Every section is equipped with fully automated instruments and semi automated
instruments as backup.
 It is running round the clock for cardiac patients.

Sections:
 OPD Reception (Phlebotomy)
 Computer section and main reception
 Hematology lab.
 Clinical Chemistry lab. (routine and special Chemistry)
 Microbiology lab.
 Operation theater lab.

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CHAPTER NO. 2 LAB ETHICS

Lab Ethics:
Every student and researcher working in the lab should have to follow “Lab. Standard Operating
Procedures” which include personal safety, environmental safety and disposal of biological
wastes.

Personal Safety
 Must wear a buttoned lab coat. If forget your lab coat, lab coats are available in the lab,
must sign out and sign in when returning
 Lab. Coat Laundry guidelines: wash separately from other clothes with detergent and
bleach. When taking lab coat home for washing, carry separate from all other personal
effects, i.e. not in your back pack.
 No personal effects (this includes outer clothing and back packs) are permitted in the lab,
only essential lab supplies.
 Long hair must be tied back. Keep your hands away from your hair.
 Wash hands with antibacterial soap for minimum of 30 sec. before leaving the lab. Use
the hand wash sink located at safety station. Use wrists (wrist levers for your safety) to
turn the water on and off, not your hands.
 Remove gloves using finger of opposite hand to peel off other glove by inserting at wrist,
rolling off glove. Repeat with other hand. Dispose off gloves in Petri plate containers.
 No eating or drinking in the lab
 Never mouth pipette. Always use a pro-pipette.
 Cover any cuts with a bandage (first aid kit at Safety Station, hand wash sink area).
 Students are advised to wear closed toed shoes.

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Lab. Environment:
 Wash bench area before and after with disinfectant. Never assume that the lab bench has
been cleaned by the previous students.
 First aid kit present in lab Location is safety station (hand wash sink area) drawer.
 Know location of exits, fire extinguisher, eye wash, full body shower.
 Know how to operate equipment before use. DO NOT use equipment unless you know
exactly how to operate the equipment. The Teacher Assistant is always available to
assist.
 Leave your bench area clean. All equipment and supplies should be returned to original
location.

Disposal:
 All biohazard disposable containers must be labelled with a biohazard label. After
autoclaving the biohazard label is removed.
 All biohazards must be autoclaved. Biohazards include any surface that has come in
contact with bacteria.
 Discard all non-sharp biologically contaminated items in the Petri plate container -only
agar culture plates, disposable gloves and contaminated paper towels.
 Any ‘pointy’ item must be disposed in plastic lined bucket (not Petri Plate container), this
includes pipetman tips, disposable cuvettes, slides, broken glassware, metal objects.
 Biological Spills:
Put on gloves. Cordon off area to let aerosol settle for 10-30 minutes, then wash container
or area with disinfectant by putting stack of paper towels on top of spill, pour disinfectant
around and over. Do not press down. Collect soaked paper towels in Petri plate container.

 Organic solvents must be disposed off in organic solvent container. Never pour solvents
down the sink. Use extreme care with flammable solvents.
Dispose off all sharps (needles, syringe tops, razors, scalpel blades) in specified container
(red or yellow .Do not replace the needle cap before disposing.

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CHAPTER NO. 3 PHLEBOTOMY AND MAIN RECEPTION
OPD Reception (Phlebotomy):

The section for phlebotomy is in out patient department (OPD) of hospital. The OPD is the
mirror of hospital which reflects the functioning of hospital being the first point of contact
between the patient and the hospital staff.
The blood sample is collected according to “Standard Operating Procedures (SOPs)”.
It is supervised by Mr. Javaid (Phlebotomist) who gives training to the students for standard
blood sample collection.

Phlebotomy:
Phlebotomy is the practice of drawing blood from patients and taking the blood specimens to the
laboratory to prepare for testing.
The blood is usually taken from vein on the back of the hand or just below the elbow. Some
blood tests, however, may require blood from an artery.

Procedure:

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  Collect all the equipment needed for the procedure which includes:
o Sterile glass or plastic tubes with rubber caps
o Sterile gloves
o Blood sampling devices
(needle/syringe)
o A tourniquet
o Alcohol swab
o Laboratory forms
o Puncture resistant sharp containers
 Clean your hands with sanitizer.
 Ask the patient to state his/her first and last
name.
 Check the requisition form for requested tests, other patient information and any special
draw requirements. Gather the tubes and supplies that you will need for the draw.
 Position the draw site for
best visualization and/or
palpation. Apply the
tourniquet 3-4 inches
above the selected puncture
site.
 Do not place tightly or leave
on for more than 1 minute.
Instruct the patient to make
a fist.Select the venipuncture
site by palpating with the
gloved index finger.
 Wipe the skin with an
antiseptic over the area of venipuncture.
 Grasp the patient’s arm firmly using your thumb to draw the skin taut and anchor the
vein.
 Swiftly insert the needle through the skin, bevel side up, at a 15 – 30 degree angle with
the skin, into the lumen of the vein. Draw appropriate amount of blood.
 Release the elastic band and withdraw the needle from the vein.
 Draw the blood into respective vial.
Note: If the patient complains of “shooting, electric-like pain, or tingling or numbness proximal
or distal to the puncture site,” the needle should be removed immediately. It is possible that a
nerve has been punctured and possibly damaged. The venipuncture should be repeated in a
different site.

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Sites for Venipuncture:

Preventions:
Preventing Hematoma:
 Puncture only the uppermost wall of the vein.
 Remove the tourniquet before removing the needle.
 Make sure the needle fully penetrates the upper-most wall of the vein; partial penetration
may allow blood to leak into the tissue surrounding the vein.
 A hematoma can cause a post-phlebotomy compression injury to a nerve.
Preventing Hemolysis:
 Mix tubes gently, by inversion, 5-10 times, do not shake them.
 If using a needle and syringe, avoid drawing the plunger back too forcefully.
Preventing injury to a nerve, tendon, or muscle:
 Use careful palpitation and appropriate angle of entry. Avoid excessive probing.
Preventing infection:
 Follow proper infection control policies.

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Sites to be avoided for blood collection:

Computer section and main reception:

This is the main section of laboratory where the samples are received and reports are delivered to
the patients. After receiving the samples:
 Checking sample integrity and deciding if the sample is fit for analysis or not.
 The data is entered into the laboratory computer.
 The samples are labeled and then the data is entered into the computer system.
 Finally, the samples are dispatched to the respective departments.

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CHAPTER NO. 4 HEMATOLOGY

The test performed in hematology section are:

 Complete Blood Count (CBC)
 Erythrocyte Sedimentation Rate (ESR)
 Prothrombin Time ( PT-INR)
 Activated Partial Prothrombin Time (APTT)

Complete Blood Count (CBC):

The complete blood count (CBC) is one
of the most common blood test used.It is
a blood test used to evaluate your overall
health and detect a wide range of
disorders, including anemia, infection
and leukemia.
The machines used for CBC is:
o Sysmax KX-21

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Following are the parameters of CBC:
Parameters Normal ranges

ESR 0-18
Total WBCs 4000-11000/mm3
Total RBCs 4.5-6.5/mm3
Hemoglobin 14-17.5g/dL (For Male)
12.3-15.3g/dL (For Female)
Packed cell volume 40-54%
Mean cell volume 76-96%
Mean cell hemoglobin 20-32%
Mean cell hemoglobin conc. 30-35%
Platelet count 150000-400000/mm3
Neutrophils 50-70%
Lymphocytes 20-50%
Monocytes 1-6%
Eosinophil 0-6%
Sample requirement:
The sample should be in CBC vial having ethylene-diaminetetraacetic acid (EDTA). The clotted
sample will not be acceptable.
o 1.6ml blood sample
o 0.4ml EDTA

Procedure:
The test is performed on fully automated machine.
 Place the samples on the roller mixer for complete mixing of the sample, as it is
necessary so that all the blood components are mixed and result may not disturbed.
Mixing of sample is done for at least 5-10 min.
 Press sample No. button.
 Enter the sample No. then press enter.
 Put the sample vial below sample probe and press start button.
 Remove the sample when analyzing is displayed.
 Wait for result. The print will come out automatically.

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Results:
Cells Causes of low count Causes of high count

WBCs Bone Marrow Damage, Viral Infection,

Autoimmune disease, Inflammation,
Disease of Immune Asthma
System e.g. HIV/AIDS
Eosinophils Numbers are normally low in Parasitic Infection,
the blood. Medically Addison disease,
insignificant
Basophils Numbers are normally low in Allergic reactions,
the blood. Medically Inflammation,
insignificant Uremia
RBCs Acute bleeding, Dehydration, Lung Disease,
Nutritional deficiency, Living high altitude
Bone Marrow damage
Mean cell volume Indicates RBCs are smaller Indicates RBCs are larger
than normal (microcytic); than normal (macrocytic),in
caused by iron deficiency anemia caused by vitamin
anemia B12 or folate deficiency,
hypothyroidism
Mean cell hemoglobin conc. Iron deficiency, anemia and autoimmune
thalassemia. hemolytic anemia
Platelets Viral Infection, Cirrhosis Cancer, Arthritis

Neutrophils Nutritional deficiency, Acute infection

Tuberculosis ,
Dengue fever,
viral hepatitis
Lymphocytes infectious diseases , Acute lymphocytic leukemia,
autoimmune disorders, blood Chronic lymphocytic,
cancers and blood diseases leukemia,
Whooping cough,
Tuberculosis.

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Erythrocyte Sedimentation rate:
The erythrocyte sedimentation rate (ESR) is a relatively simple, inexpensive, non-specific test
that has been used for many years to help detect inflammation associated with conditions such
as infections, cancers, and autoimmune diseases.

The machine used for ESR is HUMASED 20.

Procedure:
 In laboratory the sample test tube is placed on the roller
mixer for complete mixing of the sample, as it is
necessary so that all the blood components are mixed
and result may not disturbed. Mixing of sample is done
for at least 5-10 min.
 Draw the blood into the ESR tube (having sodium citrate
3.8%) up to the mark.
 Place it into the machine and wait for the results.
 Note the results.
Normal range: 0-18 mm/hr

PT-INR and APTT Test:

The prothrombin time (PT) is used, often along with activated partial thromboplastin time (PTT),
to help diagnose the cause of
unexplained bleeding or inappropriate
blood clots.
The international normalized ratio
(INR) is a calculation based on results
of a PT and is used to monitor
individuals who are being treated with
the blood-thinning medication.
The machine used for this test is
Humaclot Duo plus Human.

PT-INR Procedure:
 Centrifuge the sample for about 15 minutes.
 Place the sample in the water bath at temperature of 37ºC.
 Add 100 microliters PT reagent.

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  Add 50 microliters plasma.
 Note the results.
Normal range: 10-14 seconds.

APTT Procedure:
 Centrifuge the sample for about 15 minutes.
 Place the sample in the waterbath at temperature of 37 degree celcius.
 Add 100 microliters APTT reagent.
 Add 100 microliters plasma.
 Wait for about 2 and half minutes.
 Then add CaCl2.
 Note the results.
Normal range: 30-34 seconds.

Peripheral blood film:

A blood film or peripheral blood smear is a thin layer of blood smeared on a glass microscope
slide and then stained in such a way as to allow the various blood cells to be examined
microscopically. Blood films are examined in the investigation of hematological (blood)
disorders or malaria parasite.

Requirements:
 Blood samples drawn in EDTA having vial
 Glass slides
 Dropper
 Giemsa stain and methanol

Procedure:
 Place a drop of blood on one end of a slide
 Place the spreader slide at an angle of 30-45° near the drop of blood.
 Spread the drop by making a monolayer.
 Air dry the slide and after that fix the slide by immersing it briefly in methanol.
 Finally, stain it by Giemsa stain.

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CHAPTER NO. 5 CLINICAL BIOCHEMISTRY

Clinical biochemistry is the area of clinical pathology that is generally concerned with analysis
of bodily fluids for diagnostic and therapeutic purposes, mostly on serum or plasma.
Routine Chemistry Special chemistry
 Blood glucose  Troponin I
 Renal function tests  T3,T4,TSH
 Liver function tests
 Lipid profile
 Cardiac enzymes
 Serum electrolytes

Routine chemistry:
1. Blood Glucose:
Principle: This test is done to check glucose level in the blood.
Normal range: Fasting blood glucose less than 100 mg/dL
Random blood glucose below 125 mg/dL

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2. Renal Function Test:

Tests Principle Normal ranges

Blood urea nitrogen Measures the amount of Adults7-20 mg/dl
(BUN) nitrogen in the blood that Children5-18 mg/dl
comes from waste product Patients on dialysis40-60
urea. High BUN level mg/dl
indicates kidney
dysfunction

Serum creatinine Measures the blood level of 0.5 – 1.1 mg/dl

creatinine. Its high level
suggests kidney problem.
Uric acid Helps to diagnose gout and 3.5 – 7.0 mg/dl
kidney stones.

3. Liver Function Test:

Tests Principle Normal ranges

SGPT/ALT It is an enzyme found in 0 – 41u/l
muscles and many other
tissues besides the liver.
Alkaline phosphatase: when bile flow is slow or 40 – 129 u/L
blocked, the level of
alkaline phosphatase will
increase which indicates
liver problem
Serum albumin Measures the amount of 3.4 – 5.5 g/dL
albumin in the serum. Its
low level indicates kidney
or liver disease
Total bilirubin Measures jaundice, liver 0.1 – 1.2 mg/dL
disease and blockage of
bile duct
Direct bilirubin Estimates the amount of 0.0 – 0.5 mg/dL
conjugated bilirubin
present.
Indirect bilirubin Measures the amount of 0.1 – 0.7 mg/dL
bilirubin attached to
protein.

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 4. Lipid Profile:
Tests Principle Normal ranges
Cholesterol Measures the total amount 140 – 200 mg/dL
of fatty substances in blood.
HDL Estimates the level of good 35 – 65 mg/dL
cholesterol in blood.
LDL Estimates the level of bad Less than 100mg/dL
cholesterol in blood.
Triglycerides Measures the total amount 50 – 150 mg/dL
of fatty substances in blood

5. Cardiac Enzymes:
Tests Principle Normal ranges
CK-MB (Creatine Indicates heart or skeletal 0 – 25 u/L
kinase myoglobin) muscle damage. Increased
CK-MB can usually be
detected in someone with
heart attack about 3-6 hours
after chest pain.
CPK (Creatine When a muscle is damaged, 24 – 245 u/L
phosphokinase) CPK leaks into the blood
stream. Form of CPK helps
to determine which tissue
has been damaged
SGOT/AST Its elevated level indicates 8 – 40 u/l
heart damage.

All these tests are performed in 902 automatic chemistry

analyzer HITACHI.
 Firstly the samples are centrifuged.
 Pour 30 µl serum in sample cups.
 Start entry and select required test.
 Place sample at respective places.
 Start the machine and wait for the results.

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Special Chemistry:
1) Troponin I:
This test is done to diagnose the heart attack and rule out other conditions with signs and
symptoms.
Troponin I is the protein that is found in hearts muscle and are released into the blood
when there is damage to heart.
Normal range: less than 0.01 ng/mL

Procedure: (Manual)
 Centrifuge the sample.
 Remove the testing device from the pouch and
place the device on bench.
 Pipette drop wise 3 free falling drops of undiluted
fresh serum sample into the sample well of test
device.
 Place the sample in Biocard reader device
immediately after 12 minutes.
 Give the sample ID and start the machine.
 Wait for the results and print.

2)T3,T4,TSH:
This test is done to diagnose the thyroid problems.
Normal ranges: T3 0.5 – 1.8 ng/ml
T4 5.0 – 13.0 ng/ml
TSH 0.5 – 5.5 IU/ml
High T3 level indicates Graves’ disease, hyperthyroidism, painless (silent) thyroiditis,
toxic nodular goiter.
Low T3 level indicates hypothyroidism or starvation.
High T4level indicates iodine induced hyperthyroidism, subacute thyroiditis.
Low T4 level indicates iodine deficiency and hypothyroidism.

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Serum Electrolytes:
This test evaluates symptoms of heart disease and monitor the effectiveness of treatments for
high blood pressure, heart failure and liver and kidney disease.
The machine used for electrolytes is HUMALYTE PLUS3HUMAN.

Procedure:
 Press the sample No. button.
 Give the sample ID.
 Press OK.
 Put the serum below the sample probe.
 Machine will aspirate the serum.
 Wait for the result analysis.
 Print will come out automatically.
Normal Ranges
Na: 135 – 145 mmol/L
K: 3.5 – 5.5 mmol/L
Cl: 98 – 108 mmol/L

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CHAPTER NO. 6 MICROBIOLOGY

The tests performed in microbiology lab are:

 Blood culture
 Urine culture
TESTS FOR SEROLOGY
 Swab culture  Anti HCV.
 Sensitivity Testing
 HBsAg.
 Catalase
 Coagulase  ASO titer
 Oxidase  RA factor
 Fluid complete examination  Typhidot.
 Cell counting in Neubauer Chamber
 Urinalysis
 Staining

Cultures:
The procedure of growing microorganisms or other living matter in a specially prepared nutrient
medium.

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 1) Blood culture:
Blood cultures are used to detect the presence of bacteria or fungi in the blood, to identify the
type present, and to guide treatment.
Types of agar used:
 Blood agar
 Macconkey agar
 Mannitol salt agar
Procedure:
 Sample is taken into the culture bottles and transport to the lab.
 Incubate the sample for about 24
hours.
 Incubate the agar plates at 37oC.
 Apply one drop of blood on agar
plate, red hot the loop and spread in
zigzag direction with the help of
loop.
 Incubate the plates.
 Read the culture after 24 hours and
record the observations.
 If growth is not present then proceed to second subculture. If growth is present the make
the slide or smear.
1) Urine Culture:
A urine culture is a test to find and identify germs (usually bacteria) that may be causing a
urinary tract infection (UTI).
Type of agar used:
 Cystine lactose electrolyte deficient (CLED) agar
Procedure:
 Incubate the agar plate at 37oC.
 Spread urine on plate and discard the extra urine.
 Incubate the plates.
 Read the culture after 24 hours and record the observations.
2) Swab culture:
The sample for swab/pus culture is taken on swab from the site having pus.

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Types of agar used:
 Blood agar
 Macconkey agar
 MSA….
Procedure:
 Incubate the agar plate at 37oC.
 Apply/ rub the swab on plates in
zigzag
 Incubate the plates.
Read the culture after 24 hours and record the observations.

Antibiotic Sensitivity :
An antibiotic sensitivity test is done to help choose the antibiotic that will be the most effective
against the specific types of bacteria or fungus infecting the patient.
Type of agar used:
 Muller Hinton agar
Procedure:
 Take 1.5cc normal saline in test tube.
 Red hot the Nikon loop, take the sample
on it from the culture plate.
 Mix the colony in the normal saline with
the help of loop.
 Pour it on the culture plate and spread it
on whole plate and discard the extra.
 Incubate it for 45 minutes.
 Then apply the antibiotic discs on it
carefully and incubate, note the results
after 24 hours.
 The zone of inhibition shows the
susceptibility of bacteria to that antibiotic.

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Biochemical Tests:
The biochemical test performed in FIC are:

 Catalase, Coagulase, Oxidase

1) Catalase Test:
This test is done to identify the catalase producing bacteria such as Staphylococcus aureus.
Procedure:
 Pour a drop of hydrogen peroxide on the slide
and mix the growth in it.
 If the bubbles are formed it means the test is
positive.
2) Coagulase Test:
This test is done to differentiate Staphylococcusaureus
(positive) from S.epidermidis and S. saprophyticus (negative).
Procedure:
 Put a drop of normal saline on the slide and
mix the growth in it.
 Add two drops of fresh plasma in that.
 If result is positive agglutination will occur.

3) Oxidase Test:
The oxidase test is a test used in microbiology to determine if a bacterium produces certain
cytochrome c oxidases such as Pasteurella species., which cause metabolic disorders.
Procedure:
 Take a bacterial colony on pipette tip and rub
it on the oxidation paper.
 If test is positive, purple color will appear.

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Fluid complete examination:
The fluid may be:
 Pericardial fluid.
 Cerebrospinal fluid.
 Peritoneal fluid.
 Pleural fluid.
3 basic steps:
Physical Examination:
Volume, Color, turbidity, coagulum, deposits.
Chemical Examination:
Glucose, total protein, albumin, LDH.
Microscopic Examination:
RBCS, WBCs, total cells, lymphocytes, polycytes, AFBs, morphology ofcells, gram
positive or negative bacteria.
 Microscopy is done with Neubauer chamber (hemocytometer) or stained slides.

Neubauer Chamber:
 Make the dilution of blood
sample.
 Place cover slip over the chamber.
 The tip of the pipette is placed
near the cover slip.
 Capillary action will draw the fluid
into the chamber
 It is important not to overload the
chamber, as doing so will give an
inaccurate count.
 Count the cells by applying formula:
Total cells = No. of cells x 2.5 x dilution factor

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Urinalysis:

Urinalysis is used to detect and assess a wide range of disorders, such as urinary tract infection,
kidney disease and diabetes. Urinalysis involves examining the appearance, concentration and
contents of urine.
3 basic steps.
Physical Examination:
Color, turbidity, deposits.
Chemical Examination:
Specific gravity, pH, glucose, ketones, blood, bilirubin, protein, nitrite.
Microscopic Examination:
RBCS, WBCs, Crystals, Epithelial Cells, Casts.

PROCEDURE:
 Number the urine samples, test tubes and
payment slips.
 Check the physical parameters and note
them down on slip.
 Dip the strip for chemical examination in
sample, take it out and note the results
after 60 sec. by matching color.

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  Pour the sample in test tube and centrifuge.
 Discard the sample and the remaining is used for microscopy.
 Label the slides, put a drop of sample on slide and place cover slip on it.
 Examine the slide under the microscope and note the results for microscopy.
Urine for ketone bodies:
 Ketone bodies are present in the case of diabetic ketoacidosis.
 High glucose conc. in the body accelerates the metabolism due to this increase
acetoacetic acid is produced.
 Osmotic diuresis occurs which leads to dehydration.
 Breathing rate is also increased.
 All these conditions together leads to coma.
 The procedure is same as urine for C/E.

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Staining:

The process of impregnating a specimen for microscopic examination with coloring matter that
matter that makes it visible.
There are three types of staining:
 Gram Staining
 To view gram positive or gram negative bacteria
 Giemsa Staining
 To view morphology of bacteria or cells
 Zeihl-Neelsen Staining
 To view acid fast bacteria

Gram Staining:
 First of all, make the smear and let it to dry.
 Heat fix the smear.
 Staining with crystal violet for 1-2min
 Wash
 Pour iodine solution for 1-2 min
 Wash
 Decolorize with acetone alcohol for 5-10 sec
 Wash
 Pour safranin for 1-2 min
 Wash and dry.

Giemsa Staining:
 First of all, make the smear and let it to dry.
 Fix the slide with methanol.

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  Make 1:3 dilution of giemsa stain.
 Pour the stain over the smear for 14-20 min.
 Wash and air dry the slide.

ZN Staining:
 Heat fix the smear.
 Carbol fuchin heat until vaporization then wait for 5min.
 Wash.
 Decolorize with 25% H2SO4 for 10 sec.
 Wash.
 Pour methylene blue for 3min.
 Wash and dry.

Serology:

Serology is the scientific study of serum and other bodily fluids. This test is done to detect the
presence of antibodies against a microorganism.

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Anti-HCV and HBsAg:
Anti HCV and HBsAg kits are used for test. (Chromatographic method)

Anti HCV HBsAg

 Works on the principle of sandwich  Works on the principle of direct
ELISA ELISA.
 Centrifuge the sample.  Centrifuge the sample.
 Remove the testing device from the  Remove the testing device from the
pouch and place the device on bench. pouch and place the device on bench.
 Pipette one drop undiluted fresh serum  Pipette drop wise 3 free falling drops
sample into the sample well of test of undiluted fresh serum sample into
device. the sample well of test device.
 Pipette two drops of buffer which act  Result will be shown by antigen
as secondary antibody. antibody reaction.
 Results will be shown by antigen
antibody reaction.

Antistreptolysin O (ASO) titer:

It is a blood test to measure antibodies against streptolysin O, a substance produced by group A
Streptococcus bacteria which causes glomerulonephritis or rheumatic fever in a person with
signs and symptoms.

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The amount of ASO antibody (titer) peaks at about 3 to 5 weeks after the illness and then tapers
off but may remain detectable for several months after the strep infection has resolved.

Procedure:
 Pipette 50µl drop of serum on slide. Then add 1 drop of ASO titer reagent on it.
 Mix it in circular motion.
 If there is agglutination, the test will be positive and the value will be >200.
 If there isno agglutination, the test will be negative and the value will be <200.
 To calculate the exact value, make the dilution 1:2 of serum with normal saline.

 Repeat the procedure.

 If the result will be positive then go for next dilution that is 1:3.
 If the result will be negative then multiply the dilution factor with 200.
 Note the results.
ASO titer value = dilution factor * 200

RA factor (Rheumatoid factor):

These are proteins produced by your immune system that can attack healthy tissue in your body.
High levels of RA factor in the blood are most often associated with autoimmune diseases, such
as rheumatoid arthritis.

Procedure:
 Mix blood with tiny rubber (latex) beads that are covered with human antibodies.
 If RA Factor is present, the latex beads clump together.
 Agglutination will occur.

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Typhidot test:
Typhidot is a medical test consisting of a dot ELISA kit that detects IgM and IgG antibodies
against the outer membrane protein of the Salmonella typhi.
The typhidot test becomes positive within 2–3 days of infection and separately identifies IgM
and IgG antibodies.

Procedure:
 Centrifuge the sample.
 Remove the testing device from the pouch and place the device on bench.
 Pipette drop wise 3 free falling drops of undiluted fresh serum sample into the sample
well of test device.

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 OPERATION THEATER LAB
The tests performed in FIC OT-Lab are:
Arterial Blood Gases
 Arterial blood gases
 pH
 Sodium
 pO2
 Potassium
 pCO2
 Glucose
 O2 saturation
 Hemoglobin
 Bicarbonate
Arterial blood gases:
An arterial blood gas (ABGs) test measures the acidity (pH) and the levels of oxygen and carbon
dioxide in the blood from an artery. This test is used to check how well your lungs are able to
move oxygen into the blood and remove carbon dioxide from the blood.
The machine used for ABG’s is COBAS B 121.

Procedure:
 Take heparinized whole blood sample and
check for clot, air bubbles.
 Put off cap of syringe and discard air bubbles
and discard few drops.
 Enter the patient name.
 Press OK and lift the flap of sample probe side.
 Insert the sample probe into nozzle of syringe
in blood sample.
 Remove the syringe when the beep sound is
heard.
 Wait for 1 to 2 minutes, the result will come out
as print automatically.

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Learning Outcomes:
In a nutshell, this internship has been an excellent and rewarding experience. One main thing that
I have learned through this internship is time management skills as well as self-motivation. I
have gained new knowledge, skills and met many new people. I have learned and practiced all
the methods and procedures that are performed in the Pathology Lab. and also came to know the
normal ranges and values that will help in diagnosis of disease.
The internship was also good to find out what my strengths and weaknesses are. This helped me
to define what skills and knowledge I have to improve in the coming time. At the beginning of
the internship I formulated several learning goals and I have achieved many of them which
includes:
 Understand the functioning and working condition of a Government organization.
 Practical implementation of theoretical study.
 Experience of professional field.
 Use my gained skills and knowledge.
 To see what skills and knowledge I still need to work in a professional field.
 To buildup self confidence.

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