Indications of Frozen sections.

1) A Urgent diagnosis during surgery (Benign and Malignant)

2) Involvement of resection margins by, malignancy.Ex: basal cell carcinomas.
3) Ganglion cells in Hirschprung disease…
4) Enzyme histochemistry.
5) Non enzyme histochemistry.
6) For DIF Silver and gold impregnation methods.Identification of the type of tissue. Ex: Parathyroid
gland.

Mention various mounting solutions used in museum specimen.

 Ans: Mounting solution

 Formalin 400 ml
 Water 2000 ml
 Potassium Nitrate – 30 gm
 Potassium acetate – 60 gm
 80% Ethyl alcohol.

Screening test for blood donors.

 Ans: Quality assured screening of all donated blood for transfusion.

 Transmissible infections-HIV, Hepatitis-B
 Hepatitis-C, Treponemapallidum.
 Serologic test for syphillis.
 The purpose of donor screening and referral procedures is to minimize the possibility, of
transmitting an infectious agent from a unit of donated blood to recipient of the unit as well as
ensuring the welfare of the donor himself.

Microscopic examination of urine.

 Ans: Microscopic urine analysis identification of formed elements present including casts, cells,
crystalls (3 Cs), micro-organisms mucus and arefacts.
 Procedure:
1) Use a clean, fresh early morning specimen.
2) Obtain urinary sediment by centrifuging urine at 3000 rpm for 5 minutes.
3) Draw off the clear supernatant fluid.
4) Place a drop of the sediment-on the glass slide and cover it with a cover slip.
5) Examine first under low power and then under high power, vary the light intensity for screening
casts

Different-types of Haematoxylins.

 Ans: Types of Haematoxylins :

 Alum haemotoxylin
 Iron haemotoxylin
 Tungsten haemotoxylin
  Molybdenum haemotoxylin
 Lead haemotoxylin
 Haematoxylon without moderant.

Transfusion reaction.

 The most common reaction of blood transfusion is fever, chills and urticaria.
 It include acute and delayed heamolytic transfusion reactions and bacterial contamination of
blood product.
 Acute Haemolytic transfusion reaction.
 Fibril reactions.
 Allergic sections.
 Transfusion transmitted infection.
 Transfusion related acute lung injury.
 Delayed heamolytic transfusion reaction

Separation of components.

 From the blood donation we get blood known as whole blood.

 Blood contains cellular components and liquid (plasma).
 Cellular components are RBC, WBC, pH.
 Plasma components are FFP (fresh Frozen plasma) or cryopeptitate.
 Blood components the various constituent separated by whole blood are in blood components.

Staining of bone marrow slides.

 Wright’s or wright – Giemsa stains are usually the preferred staining method for bone marrow
aspirate smears solution with similar dye composition to the cliff- quick stain but require longer
stain contact time for adequate staining.

Causes of neutrophilia.”

 Some viral infection

 Some fungal infection
 Some parasitic infection
 (eg. Hepatic amoebiasis pneumocystis carinii)
 The commonest pathological causes in pyogenic bacterial infection.

Laboratory Information System (LIS).

 Ans: Laboratory information system is computer software that processes, stores and manages
data from all stages of medical processes and tests.
 Physician and lab technician use laboratory information systems to co-ordinate varieties of
inpatient and outpatient medical testing, including hematology, chemistry, immunology and
microbiology. Basic laboratory information systems commonly have features that
manageypatient check in order entry, specimen processing, results entry and patient
 demographics. An LIS tracks and stores and clinical details about a patient during a lab visit and
keep the information stored in its database for future reference.

Normal values of DC.

 Neutrophil : 45-65%
 Eosinophil: 1-4%
 Basophil : 0-1%
 Lymphocyte: 25 – 45%
 Monocyte : 1-8%

Tissue Embedding.

 Ans: Embedding is the process in which the tissues or the specimens are enclosed in a mass of
the embedding medium using a mould. Since the tissue blocks are very thin in thickness they
need a supporting medium in which the tissue blocks are embedded.
 Embedding of tissue is done in molten wax (paraffin wax 58-60°C) blocks of which are prepared
using metallic L moulds.

Forward and Reverse grouping?

 Ans: Forward grouping: ABO-testing is a two- part process involving testing a persons red cells
for A and / or B antigens as well as testing the persons’s serum/ plasma for ABO antibodies.
 Reverse grouping: Cells indicates the presence of of absences of anti- A and anti- B in serum.
 Reverse grouping cells: including A, B, O and A type red blood cells (RBC) are important to
resolve AB discrepancies.

Principles of microwave tissue processing.

 Microwaves are electromagnetic wave with a- wavelength shorter than a normal radio wave but
longer than infrared radiation. By using a suitably modified microwave oven, tissues can be
processed rapidly. Heat is generated which warms the tissue uniformly in a short time. This
results in faster penetration of the tissue processing chemicals into the tissues resulting in rapid
processing

Importance of bone marrow examination.

 Examination of the bone marrow is an invaluable diagnostic aid and is of value in confirming a
diagnosis suspected on a peripheral blood smear examination, which must always precede the
bone marrow examination.
 Bone marrow examination remains a simple and reliable technique in the diagnosis of many
major clinical conditions. It is an important tool for diagnosing various haematological malignant
and nonmalignant conditions.
Anticoagulants used in collecting bone marrow particies.

 Definition: A chemical used to prevent the formation of blood clots.

 EDTA: Ethylene Diamine Tetra acetic acid.
 To collect bone marrow into a syringe that contains EDTA as an anticoagulant, because it reduces
the chance of clotting before a smear is prepared and allows time to prepare multiple smears
that may be needed for special stains.

Tissue Processing.

 Ans: Fixation is the first step and should be complete and adequate.
 Processing is the procedure following fixation which makes the tissues suitable for embedding.
Each of the steps of the processing method involves the diffusion of a solution into tissue and
replacement of the previous solution.
 Dehydration :
 Dehydration is done to remove fixative and water from the tissue and replace them with the
dehydration fluid.
 Types of dehydrating agents: Ethanol, Esopropyl alcohol, Acetone.
 Clearing is replacing the dehydrating fluid with a fluid that is totally miscible with both the
dehydrating fluid and the embedding medium..
 Clearing agents: Xylene, Toluene, Chloroform, Benzene.
 Impregnation: is the process of complete removal of
 Clearing agents and substitution by wax like paraffin in order to make the tissue suitable for
embedding.

Sperm Count.

 Ans: Sperm count : A count of the number of sperm in sample of semen. A

sperm count may be used as a measure of fertility.
 A Total number of sperm in an ejaculation.
 It is 20 million / ml i.e. 60 million / ejaculation.
 It is obtained by multiplying the sperm concentration by the volume.
 Azoospermia: No spermatocytes (male)
 Oligospermia: < 20 million / ml less than 50 million / ejaculation.
 Polyzoospermia : . May reach 350 millions / ejaculation

How to prepare Leishman’s stain?

 Dissolve 0.15 g of Leishman’s stain powder in 100 ml of absolute methanol.

 The stain is then filtered into stock bottle.
 Place at 50°C for 15 minutes in water bath.
 Again filter into clean brown borosilicate glass bottle and store in a dark room temperature.
Synovial Fluid.

 Ans: Synovial fluid is the clear, pale yellow fluid the is contained in every joint in the bodies.
It is derived from plasma, which is the protein salt solution that makes up the liquid portion
of blood.
 Synovial fluid contains large amount of hyaluronic acid, which help to make the fluid more
viscous or thicker

Microscopic examination of sputum.

 Ans: Microscopic examination:

 Smears: 2-3 direct smears are made on a clean dry glass slides.
 Stain used: Leishman stain for differentiate count.
 Cells: Normal sputum consists of neutrophils, few lymphocytes, and carbon laden
macrophages (dust cells).
 Pus cells (Neutrophils): Numerous pus cells indicate pyogenic (pus-forming) infection.
 Eosinophils Increased eosinophils are seen in asthama and parasitic disease.

Fixatives.

Ans : Physical methods :

 Heating, Microwaving, Freeze drying.

Chemical methods:

 Coagulant fixatives
 Alcohol and acetone
 Picric acid and trichloro acetic acid.

Cross linking fixative :

 Formaldehyde, Other aldehydes ,Metal salts ,Component fixatives.

Uses of Neubauer chamber.

 A device used for cell counting.

 Made of special optical glass.
 Used to count cells in suspension under a microscope,
 Improved Neubauer chamber most commonly used in haematology.

Cryostat.

 Ans: A cryostat is a device used to maintain low

 Cryogenic temperature of samples or devices mounted within the cryostat.
 Use a cryostat to preserve frozen tissue samples while a microtome, an extremely sharp
cutting instrument mounted inside cryostats, slices the tissue into pipes thin enough to be
observed under a microscope
Prothrombine Time.

 Ans: Prothrombine time-is a blood test that measures how long it takes blood to clot. A
prothrombine time test can be used to check for bleeding problems. PT also used to check
whether medicine to prevent blood clot is working.
 Prothrombine time reagent.
 Contains thromboplastin and calcium chloride.

Different type of wax used in histopathology.

 Ans : Paraffin waxes that are commonly used for histological applications are straight chains
of 20-40 carbon atoms that melt in the 56-58° C range.
 Ex: Paraplast, Paramal, Polyfin, Paraffin wax.

Bar bodies.

Ans: Represent the inactive ‘X’ chromosome and are normally found only in female somatic cells.\

Sex linked inheritance :

Physically condenses to form a Barr body, a small. Structure found at the rim of the nucleus in female
somatic cells between divisions. The discovery of ‘X’ inactivation is generally attributed to geneticist
Marry Lyon, and it is called lyonization.

PAS-stain.

 Ans: is staining method used to detect poly saccharides such as glycogen, and mucosubstances
such as glycoproteins, glycolipids and mucins in tissue.
 Periodic acid-Schiff stain
 Uses:
 In diagnosis of poorly differentiated adeno carcinoma of various tissue like stomach, pancrease,
lung.
 In diagnosis of hepato cellular carcinoma.
 To demonstrate basement membrane.
 RCC-Renal cell carcinoma-clear cell type.

Blueing in H and staining.

Ans: One of the step in the H and E procedure is blueing. As the name implies, this step converts the
initial soluble red colour of hematoxylin within the nucleus to an insoluble blue colour.

Steps:

1) Remove the wax

2) Apply the Hematoxylin Nuclear stain
3) Complete the nuclear stain by Blueing
4) Remove the excess background stain
5) Apply the eosin counterstain.
 Uses of gel card in blood bank.

 Any immunohaematology test that has haemagglutination as its end point.

 ABO-Rh-typing for other blood group systems.
 Antibody screening and identification.
 Compatability testing-Cross-matching.
 DAT/IAT, other coomb’s test phase.
 Antibody-classification-IgG, IgM, IgA components.
 Specialised haematological test-sickle cell anemia.

Platelet apherisees.

 Ans: Apheresis is derived from Greek word Meaning “to take away” –
 Is a technique in which whole blood is withdrawn-separated into a components desired
component is retained and remaining constituents are returned to donor.
 Platelets are essential for blood.
 Apheresis is the process of separating blood into its different components. Platelets, Red
blood cells, and plasma.

Carbois fluid

 Solution is a substance used for as a complementary treatment after the conservative

excision of odontogenic keratocyst.
 Carnoy’s fluid is a Histological tissue fixative routinely used preserving RNA, nissl granules
and glycogen.
 Cornoy’s fluid intended for In-vitro diagnostic use. Tissue fixed with carnoy’s fluid can be
used for routine special stains and Immunohistochemical (IHC) stains.

Role of medical officer in blood bank.

 Ans: Role of medical officer in blood bank :

 Perform routine donor evaluation and monitoring.
 Physical examination and review of periodic laboratory testing.
 Provide consultation to blood bank technical and clerical personnel concerning donor selection
and acceptability.
 Evaluate and manage blood donor reactions.

Anti-coagulants in blood bank.

 Parenteral anticoagulants
 Heparin, Fondaparinux ,Danaparoid
 Oral anticoagulants
 Bishydroxycoumarin, Warfarin sodium ,Acenocoumarol ,Phenindione

APTT

 Ans: Activated Partial Thromboplastin Time is a screening test that helps evaluate a person
ability to appropriately form blood clots.
  It measures the number of seconds it takes for a clot to form in a sample of blood after
substances (reagent) are added.

Transfusion reactions.

Ans: Transfusion reaction is a serious complication that can occur, after a blood transfusion. The
reaction occurs when the red blood cells that were given during the transfusion are destroyed by the
person’s immune system. When red blood cells are destroyed, the process is called hemolysis.

Clot Retraction time.

 Ans: An old test of platelet function, not done now days.

 Clot refraction is a function of the refractile protein, thrombosthenin, present in blood
platelets.
 Take 3-5 ml of blood in a test tube allow it to clot.
 Place in a water bath at 37°C.
 Leave it undisturbed.
 At the end of 1.2.4.12 and 24 hours observe the clot and record the finding.
 The clot should be firm and refracted from the sides of the tube, and occupy half the original
volume.

Name decalcifying fluid.

 Neutral ethylene diamine tetraacetic acid (EDTA) decalcifying solution.

 5% nitric acid
 Perenyi’s fluid
 Formalin-nitric acid
 5% trichloracetic acid-
 10% formic acid.

Normal Hemostatasis, mechanism of coagulation.

Ans: Normal Hemostasis: It is a biological or physiological phenomenon which is responsible to keep the
blood in fluid state in the circulation as well as to arrest bleeding followed by an injury to blood vessel.

Mechanism of coagulation :

Hemostasis includes three steps that occur in a rapid sequence.

o Vascular spasm or Vasoconstriction a brief and intense contraction of blood vessels.

o Formation of platelet plug.
o Blood clotting or coagulation which reinforces the platelet plug with fibrin mesh that
acts as glue to hold the clot.

Role of Technician in transfusion reactions:

 Test samples to screen potential donors

 Store blood
• Draws and maintain documentation and records:
 Make patient comfortable during procedure.
  Monitor vital signs.
 A Check the compatability blood before issuing it for transfusion.

Foetal haemoglobin.

 Ans: Fetal hemoglobin (HbF) is the dominant form of hemoglobin present in the fetus during
gestation.
 Is the main O, transport protein in the fetus during the last seven months of development in the
uterus and in the newborn until roughly 6 months.
 Procedure:
o Prepare hemolysate (R1) add 0.5 ml blood to 9.5 ml drapkien then mixed.
o Transfer 2.8 ml from R1 to new tube add 200 µl NaOH (2N) mixed then incubation 5-10
min at RT.
o At the end of 2 min exactly add 2 ml saturation ammonium sulfate and mixed then
incubation 5-10 min at RT.
o Filter the solution by filter paper.
o Read the filterate at 540 nm.

Clotting time.

1) Make a clean venipuncture.

2) Timing is begun when the blood first enters the syringe.
3) Draw 3-5 ml of blood.
4) Place approximately 1 ml of blood in each of three test tubes.
5) Place tubes in a stand so that they remain upright at room temperature.
6) After 5-minutes, take the first tube and gently tip it every 30 seconds to test for clotting.
7) When the first tube is clotted, record the time and start the tipping of the second tube
every 30 – seconds until it is also found to be clotted.
8) Then do the same with the third tube.
9) The time recorded for the clotting of the third tube is taken as the clotting time.

Hospital information system.

Ans: A hospital information system is an element of health informatics that focuses mainly on the
administrational needs of hospitals.

It acts as centralized data base whereby all the information related to patients, doctors, and staffs is
collected and stored. Thus health care professionals can offer a quick diagnosis by visiting a patients
health information whenever they want.

Why validation of clinical laboratory instrument is needed?

Ans: Laboratory validation is a process that is employed to ensure that laboratory test data and results
are consistent, accurate and precise.

Validation reduces the risks of non compliance with regularity agencies. It also. Can reduce compulsory
in process controls and testing validation is a means of improving procedures and final product quality.
Quality control in histopathology.

 Accurate patient identification.

 Fixation.
 Adequate processing.
 Appropriate embedding techniques.
 Microtomy.
 Unacceptable artifacts and inspection of controls to determine correctness of special stains and
immunohisto chemical methods.

Tissue Embedding.

Ans: Embedding is the process in which the tissues or the specimens are enclosed in a mass of the
embedding medium using a mould. Since the tissue blocks are very thin in thickness they need a
supporting medium in which the tissue blocks are embedded.

Embedding of tissue is done in molten wax (paraffin wax 58-60°C) blocks of which are prepared using
metallic L moulds.

Separation of components.

 Ans : From the blood donation we get blood known as whole blood.
 Blood contains cellular components and liquid (plasma).
 Cellular components are RBC, WBC, pH.
 Plasma components are FFP (fresh Frozen plasma) .
 Blood components the various constituent separated by whole blood are in blood
components.

Histokinete- Merits and Demerits.

 Rapid penetration
 Easy availability and cheap
 Does not overharden the tissue
 Fixes lipids for Frozen sections
 Ideal for mailing.

De-Merits:

 Irritant to the nose, the eyes and mucous membranes. Formation of precipitate of
paraformaldehyde which can be prevented by adding 11-16% methanol.
 Formation of black formalin pigment, Acid formalde- hyde hematin.

Abnormalities seen in Semen Analysis

 Aspermia absence of sperm

 Azospermia – absence of sperm in semen.
 Hypospermia – Low semen volume
 Hyperspermia –High semen volume
 Oligospermia
What is Perl’s stain? Mention its use.

 Ans: Perl’s Prussian blue is a commonly used method in histology, histopathology and clinical
pathology.
 Uses:
 To detect any Ferric overload.
 Demonstrate the distribution and amount of iron deposits in liver tissue.
 Identify excess iron deposits.
 Such as hemosiderin deposits
 Hereditary hemochromatosis.

Mention 3 connective tissue stains.

 Masson’s Trichrome stain

 VAN Gieson’s stain
 Mallory’s phosphotungstic acid Hematoxylin
 Heidenhain’s Iron Hematoxyclin
 Verhoff’s Elastic stain

Mention the composition of Leishman stain.:

 Polychromed methylene blue

 Eosin azure leishman powder
 Acetone free absolute alcohol
 1.5 gms Leishman powder in 1-litre acetone free absolute alcohol.

Mention the importance of calibration of instruments in hematology.

 Calibration fine tunes Hematology analyzer and ‘provides the most accurate result purpose.
 In the normal process of tracking data for an extended period time, laboratory can make a
specific decision to recalibrate a given parameter, Never adjust to a specific value for an
individual sample.
 For best performance, calibrate all the CBC parameters.
 Ensuring correct calibration of hematology instruments can prevent physicians from making
the wrong clinical decisions.