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.