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MODEL PAPER -1
DIPLOMA IN MEDICAL LAB TECHNICIAN
SECOND YEAR
PAPER - II
Time : 3hrs. Total Marks : 80
SECTION – A
ANSWERS
SECTION – A
1. Explain about epithelical tissue?
Ans : Epithelial tissue: It covers body surfaces
and lines hollow organs, body cavities and
ducts. It allows the body to interact with
internal and external environment.
• Epithelial tissue consists of cells arranged in
continuous sheets in single or multiple lay-
ers.
• It has three major functions. They serve as:
- Selective barriers that transfer substances in
and out of the body.
- Secretory surfaces that release products pro-
duced by the cells.
- Protective surfaces.
Structure of Epithelial Tissue:
• Apical surface of an epithelial cell faces the
body surface, body cavity, lumen of an in-
ternal organ recieves cell secretions.
• The lateral surfaces of an epithelial cell fac-
ing the adjacent cells on either side may con-
tain tight junctions and desmosomes.
• The basal surface of an epithelial cell is op-
posite to the apical surface.
• Apical layer refers to most superficial layer
and basal layer is the deepest layer.
• Basement membrane is a thin extracellular
layer commonly consists of two layers. The
basal lamina and reticular lamina.
iv. Pseudo Stratified Columnar Epithelium: Function : Secretes mucus traps foreign par-
It appears to have several layers. Pseudo ticles, absorption and protection.
stratified ciliated columnar epithelium con- b. Stratified epithelium:
tain cells extend to surface and secrete mu-
i. Stratified Squamous Epithelium: It
cus.
consists of two or more layers of cells:
• Stratified squamous keratinised epithelium
develops tough layer of keratin in apical layer
of cells and several layers deep to it.
• Stratified squamous non-keratinised epithe-
lium does not contain large amounts of kera-
tin in apical layer and several layers deep
and is constantly moistened by mucus from
salivary and mucous glands.
Location : Keratinised is seen in skin. Non-
Fig : Pseudo Stratified Columnar Eoithelium
keratinised is seen in oesophagus, tongue
v. Pseudo stratified nonciliated columnar and vagina.
epithelium : It contains cells without cilia
Function: Protection, water loss and de-
and lacks goblet cells.
fence.
Location : Upper respiratory tract and epi-
didymis.
iii. Stratified Columnar Epithelium : Basal Ileocecal Valve : The lower end of the ileum
layers usually consist of shortened irregular opens on the posteromedial aspect of the
shaped cells, Apical layer has columnar cell. cecocolic junction. The ileocecal opening is
Location : Lines part of male urethra, guarded by the ileocecal valve.
conjuctiva of eye and esophageal glands.
Functions : Protection and secretion
3. Minimal or mild : When concentration, is ness is detected in the upper part, is can
less than 1 gm/24hr. sample. be due to proteins, phosphates, urates.
Causes of Proteinuria: Add 3-4 drops of CH3COOH (acetic
acid), if it disappears, it is due to phos-
i. Accidental : Due to contamination by pus, phates and urates. If it persists, it is be-
vaginal, fluid, blood or seminal fluid. cause of proteins. If mucin is present (as
ii. Physiological: Seen in pregnancy, infancy, ppt), add HN03 (Nitric acid) to dissolve
after muscular exercise, due to exposure to the precipitate. This can detect proteins
cold and sometimes relaterd to posture. as little as 2-3 mg - 100 ml.
Ortho static or postural proteinuria which is Interpretation:
usually during day when patient is upright.
1. No cloudiness-absence of proteins.
iii. Pathological :
2. Haziness-traces present (upto 10 mg/100 ml)
1. Pre-renal: No primary renal disease,
3. Cloudiness - + (10-50 mg./ml).
congestive cardiac failure, cerebral injury,
fever and toxemia, intra abdominal ascitis 4. Granular - + + (50-200 mg/100 ml).
tumours, drugs and chemical poisoning, 5. Flocculations - +++ (200-500 mg/1 100ml)
severe anaemia, plasma cell myeloma.
6. Thick cloudy precipitate - + + + +
2. Renal: Nephritis, Diabetes mellitus, SLE,
T.B of kidney, Amyloidosis, Hyperten- (> 500 mg/100 ml).
sion, Carcinoma of kidney, Uraemia of b) Contact or Heller’s Test: Take 3 ml of once.
pregnancy, HNO3 in a test tube and run urine slowly
3. Post-renal: Pyelitis, cystitis, urethritis and with the help of a pipette along the side of
prostatitis. inclined tube A sharply defined white ring
of varying density appears at the junction
Types of Proteins found in Urine: Albumin, of the two liquids If albumin is present. The
globulin, mucin, blood proteins (haemoglo- size and density of the ring can be evalu-
bin) and Bence - Jones proteins. Albumin is ated and reported as 1+, 2+. etc.,
the chief protein and is the commonest
c) Uristix orAlbustix or stix Test: A stix which is
examinatin asked for in the laboratory
already impregnated with the indicator tetra
DETECTION OF PROTEINS bromophenol biie and is dipped into urine
Principle: sample. The colour changes is from yellow
to green to blue
All methods depend on precipitation of
proteins by chemical agents or coagulation 2. Quantitative Test for Albumin
by heat. If mucin is present in excess, dilute Instrument used is Esbach’s Albuminometer.
acetic acid is added and urine is filtered. If Sample is 24 hr urine. It is a thick walled
urine is turbid, then it should be filtered as it test tube - 15 cm long and 15 mm internal
interferes with the precipitation of proteins. diameter. Markings are ‘U ’(urine) at bot-
Before starting the test, urine is made acidic tom and ‘R’ (reagent) at top. Lower part
by adding 10% of Acetic acid. has markings from 0.5 to 12 gm. The tube
is worked and stood vertically in its special
1. Semiquantitative test :
stand for 24 hours.
a. Heat and Acetic acid Test : Fill 3/4th of
Composition of Esbach’s reagent: 5 gm of
the test tube with urine and heat the upper
picric acid and 10 gm of citric acid in 500
part (lower part acts as control). If cloudi-
ml of distilled water.
Medical Lab Technician
8n
iii. Constituents of Benedict Reagent: 18 gms - Therefore, 1 ml contains 0.05/x gms is calcu-
CuS04 in 100 ml of distilled water. 100 gms lated.
- (Na 2C03) Sodium carbonate (anhy-
drous). 200 gms - Na or K citrate. This re- Ketone Bodies in urine :
sults in alkaline copper reagent. These are intermediate products of fat metabo-
iv. Advantages of Benedict’s Test: lism. They are acetone, acetoacetic acid (diacetic
acid) and B-hydroxy butyric acid. They occur
• It uses a single solution which is already in urien when fat metabolism is disordered.
prepared.
i. Rothera's Test: ( For acetone and also for
• It uses a more stable reagent. acetoacetic acid).
• It is ten times more sensitive. Procedure: For acetone and also for ac-
• It is a more specific test. etoacetic acid. Take 5 ml urine in a test tube
and saturate with Ammonium sulphate. Add
b. Fehling's Test: Take equal volumes of
1 crystal of sodium nitroprusside. Let liquid
Fehling’s solutions A and B, mix well and
ammonia run down the sides of the tube,
boil. Add a few drops of urine. Boil and ob-
so that it forms a layer on top of urine. If
serve the colour change. Colour changes
acetone is present a permanganate caramel
are same as in Benedict’s test.
red colour ring forms at the junction of the
Composition of Fehling’s solutions: two layers.
Solution A - CuSO4, in distilled water. ii. Gerhardt's test for Diacetic Acid: Procedure:
Solution B - NaOH, Sodium Potassium Take 5 ml of urine in a test tube and add
tartarate in distilled water. 10% of aqueous solution of ferric chloride
drop by drop until no further ppt of ferric
c. Test for Lactose: Rubner’s test: Concen- phosphate occurs. Filter and add to the fil-
trated NH40H is used. Lactose gives and trate more of ferric chloride (FeCIS) solu-
ppt while glucose gives a yellow ppt. tion. If diacetic acid is present, the filtrate
d. Test for Fructose: Selwinoffs test. will develop brownish red colour.
e. Test for Pentose: Bial’s test. iii. Other tests: Lindeman’s Test: For diacetic
acid.
Osazone Test: This test is used to differentiate
the various osazones. Acidified urine is used Hart’s Test: For Beta Hydroxy butyric acid.
along with phenyl hydrazine hydrochloride and Causes for presence of Ketone bodies in
sodium acetate. Glucosazone crystals are Urine:
straight, needle shaped, arranged in fans or 1. Diabetes mellitus
sheets of spherical clusters. Lactosazone crys-
tals are finely curled with puffball appearance. 2. Infants and children with acute febrile con-
ditions
Quantitative Test for Glucose:
3. Toxic states accompanied by vomiting and
Procedure: Take 25 ml of Benedict’s reagent diarrhoea
in a flask and add 15 gm of crystal Na2 C03(Na
carbonate) and some broken pieces of porce- 4. Vomiting of Pregnancy.
lain. Heat until it boils. Take urine in a burette 5. Cachexia.
and titrate against the contents of the flask. Nor-
6. Following exposure to cold.
mally, 25 ml of reagent requires 0.05 gms of
sugar for reduction. If × is the amount of urine 7. Following severe exercise.
run down, then × ml contains 0.05 gms of sugar. 8. Following anaesthesia.
Medical Lab Technician
10 n
Note : In diabetes, the high proportion of +++ - Blue.
sugar is not dangerous to the body. But, if ++++ - Deep Blue.
sugar metabolism is upset, Ketone bodies
accumulate in blood which are poisonous Other Tests:
to the body b. Guaicum test: It is less sensitive test
Blood in urine : c. Orthotolidine test
Blood may be found in urine in the form of III. Spectroscopic Examination: Look for typica.
red blood cells or blood pigments alone. It absortion bands with a hand spectroscope.
is important to indicate whether RBC are
Causes of Hematuria:
present or only pigments are present. Nor-
mally, occasional RBC may be found. a. Lesion of kidney: Cystitis, acute glomerulo-
nephritis, nephrolithiasis, urethritis, embo-
Hematuria - Denotes presence of red blood
lic nephritis, Renal TB, malignant nephro-
cells in urine.
sclerosis.
Haemoglobinuria - Denotes presence of
b. Neoplasms of kidney: Lesions in lower urinan
blood pigments in urine without associated
tract stones in bladder, urethra, Bilharziasis
presence of RBC’s
and Neoplasms.
Tests for Presence of Blood:
Bile Salts and Pigments in urine :
I. Microscopic Examination: Urine is centri-
Constituents of bile may be excreted in urine
fuged and the sediment is examined under
as bilirubin, urobilin, bile salts and bile pigments.
microscope. Report as the no. of RBC / High
Bilirubin appears in urine, when it is in excess in
Power Field.
blood i.e. in jaundice. Bile salts and pigments are
II. Chemical Tests: present in obstructive jaundice and hepatocellular
a. Benzidine Test: carcinomas.
i. Principle: The peroxidase activit of hae- Tests for Bile Salts:
moglobin decomposes hydrogen perox- Hay Sulphur Test:
ide and the liberated oxygen oxidises the
i. Procedure : Take 10 ml of urine in a test
benzidine. It is very sensitive to even small
tube and sprinkle sulphur granules. Nor-
amounts of haemoglobin.
mally they float.
ii. Procedure: To two ml of urine (previ-
In the presence of bile salts, the surface ten-
ously boiled and cooled) add one ml of
sion is reduced and the sulphur granules sink
clear saturated benzidine solution (Ben-
and settle at the bottom.
zidine powder and glacial acetic acid) mix
well. add one ml of freshly prepared 3% ii Control: Take 10 ml of water m a test tube
hydrogen peroxide. and sprinkle sulphur granules (which float)
iii. Interpretation: The appearance of green Tests for Bile Pigments:
or blue colour within 5 minutes indicate i Foarm test: Take urine in a test tube and
the presence of blood or haemoglobin shake it well. A positive result is obtained
in the urine with the formation of stable yellow froth,
Trace - Faint green Colour. the stability of which is due to the presence
of bile salts and the colour is due to biliru-
+ - Green.
bin.
++ - Greenish blue.
ii. Gmelitt’s test: Principle - Bringing yellow ni- Importance: The edge of the microtome
tric acid in contact with urine. Take 2 ml kinfe performs the function of cutting and it
yellow nitric acid in a test tube overlay with should be sharp in order to get good sections
an equal amount of urine. A band of of the tissues.
coloured rings appear, the most prominent Introduction : Sharpening of the knife can
being green colour. This demonstrates the be carried out by:
presence of bile pigments. This can be done
• Mechanically or by
on a filter paper or procelain plate.
iii. Fouchet’s test: Take 10 ml urine in a test • Automated knife charpners.
tube and add 2.5 ml of 10% BaClr Mix well The process of sharpening consists of:
and filter.
• Honing and
Then unfold the filter paper and spread on
• Stropping.
another filter and allow one drop of Fouchef
s reagent on the ppt. A green or blue colour Requirements:
indicates the presence of billirubin. This is a
• A hone (rectangular block of stone cpecially
sensitive test.
made for sharpening of knife).
Composition of reagent - Trichloroacetic
• A properly fitted back of the knife.
acid and 10% FeCl 3 in distilled water.
• Microscope to observe the edge of the knife.
Reaction :
• Knife.
Bile Salts + BaCl2 BaSQ4 ® ( ppt ) acts ®
with fouchef s reagent (colour +) • A strop (flexible or grid).
Tests for Urobilinogen: Excess can be de- Procedure (Honing):
tected by Ehrlich’s aldehyde test or by
• Clean the knife with Xylene soaked cloth.
commerical strip test.
• Put on the proper honing back.
1. Urobilineogen: On reacting with para -
dimethylaminobenzaldehyde forms a pink • Keep a damp cloth under the hone and po-
coloured compound. sition the hone on suitable bench.
ii Ehrlich’s Test: Procedure. Take 10 ml of • Lubricate the surface of hone with coconut
fresh urine in a test tube and add 2.5 ml of oil.
BaCl . Mix well and filter. Take 2.3 ml of
• Place the knife at one end and push it di-
filtrate, add 0.5 ml of aldehyde reagent and
agonally forward with the leading cutting
allow it to stand for 3 mins.
edges in the front.
Then see the top column of urine against a
• Just before the edge reaches the end of the
white background. A pink colour denotes
stone the knife is turned, over on its back
the presence of urobilinogen. Repeat the test
without lifting it. It is steadily pulled back with
with in 10, 20, 50, 100 and 200 dilutions.
the cutting edge leading again along the
The more dilute it is, the more sensivtive
hone towards the operator.
the test becomes. The report should be in
terms of highest dilution with a positive re-
action.
4. Write about sharpening of microtome
knife?
Ans : Sharpening of the Micro tome Kife:
Medical Lab Technician
12 n
• Place the knife at one end of the strop and • Tom and ragged sections are obtained and
push it diagonally forward carefully so that, even the cutting edge of the microtome knife
entire edge receives the polish. may be damaged.
• Maintain the pressure on the knife just suffi- The technique of decalcification can be
cient to keep the edge in contact with the divided into the following stages:
surface of the strop and establish a rhyth- 1. Selection of tissue.
mic, steady motion.
2. Fixation.
• When it reaches the end of the strop, turn it
3. Decalcification.
on its back and pull back towards you (to-
wards the operator). 4. Acid neutralisation and
• Observe under microscope for a sharp and 5. Washing.
even surface.
1. Selection of tissue:
a. Bone: By using fine toothed bone saw or c. 1 g /dl zinc sulfate : 0.2 ml.
hack saw, thin slices of bone are obtained. d. Chloroform : Few drops.
The slices should not exceed 4-5 mm in
Specific features:
thickness.
• Use of this buffered solution causes no per-
• The cut surfaces should be retrimmed to
ceptible damage to cells or tissue fibers.
remove damaged areas after decalcification
and washing. • The decalcification results are much slower
than other establiched fluids.
b. Calcified tissue: A sharp knife is used to cut
calcified tissue (or a saw may be used). • This solution is of use when time is not an
Duration of decalcification will depend on important factor.
the degree of calcification.
3. Jenkin's fluid
2. Fixation:Bone marrow is best fixed in Zenker
a. Absolute alcohol 73ml
formal or any other routine fixative which
may preserve the issue well, e.g. Formalin. b. Distilled water 10ml
3. Decalcification may be carried out as follows: c. Chloroform 10ml
• By using a dilute mineral acid (a simple d. Glacial acetic acid 3ml
solvent of calcium). e. Hydrochloric acid 4ml
• By forming simple combination of calcium The swelling effect of hydrochloric acid is
with ion exchange resins. counter-acted by the shrinking effect of the
• By using chelating agent, which binds the alcohol.
calcium. • Quantity of this fluid should be 40 and 50
• By removal of calcium from tissues using times the bulk of the tissue.
electrophoresis techniques. • Human rib cross sections are decalcified in
4. Acid neutralisation :Tissues should be 4-6 days
neutralised before washing by treatment, 4. Formal nitric acid:
which breal alkali such as 5g/ dl lithium or
sodium sulfate a. Formalin 5ml
5. Washing: It is carried out for 2 to 4 hours in b. Nitric acid (sp. gr. 1.14) 7.5 to 15ml
alcohol or overnight in water to remove al- c. Distilled water 100ml
kali after the neutralization.
Specific features:
Decalcifying fluids:
• Nitric acid develops yellow colour due to the
1. Gooding and Stewart fluid: formation of nitrous acid, which may inter-
a. Formic acid : 5 to 25 ml. fere with the staining reactions.
b Formalin : 5 ml. • The yellow colour can be obviate by the
addition of 0.1 g/dl urea to pure nitric acid.
c. Distilled water : 100 ml.
• Human rib cross section (5 mm) can be de-
2. Citrate-citric acid buffer: calcified in 1-2 days by using 7.5 % (v/v)
a. 7 g/ dl citric acid (monohydrate) : 5 ml. nitric acid.
b. 7.54 g/ dl ammonium citrate Electrophoretic Decalcification: It is based
on the principle of attraction of the calcium
9 anhydrous) : 9.5 ml.
Medical Lab Technician
14 n
ions in solution to a negative electrode, by • Clamping screw should be kept loose on the
passing electrical current. The electrolyte specimen head.
used in electro phoresis is equal parts of 8%
(v/v) hydrochloric acid and 10% (v/v) for- • Insert the specimen disc/ metal chuck into
mic acid. Brass plate is used as negative elec- opening of specimen head.
trode and platinum wire as positive elec- • Tighten clamping screw.
trode. The positive electrode is wound
around the specimen. This is placed in a glass • Unlock the hand wheel.
museum jar. Speed of decalcification may • Trim the specimen using coarse and preci-
be increased by moving the electrodes sion feed buttons. Use coarse feed button
closer together or decreased by moving them for moving specimen towards the cutting
further part. edge.
6. What is cryostat? • Set the section thickness to 6 to 10 microns
Ans : Working of cryostat: General Procedure: using sections thickness knob.
Fresh tissue is received for frozen section. • Place the anti-roll guide plate over the knife
All specimens should be considered holder. Sections have tendency to curl. Anti-
potentially dangerous, hence, these should roll plate supports section to form flat sec-
be handled using gloves. tions.
• Water content from tissue is absorbed by • For maintenance of cryostat use the follow-
gauze, which affects sectioning procedure. ing procedure
• Size of tissue for frozen section should be • Lock the hand wheel
about 3× 3 mm. • Remove frozen section waste with a
• A drop of cryo compound (optimum cooling branch.
medium) or water is added on the disc/ • Dispose waste as biohazard waste.
metal chuck. Cryo compound or water can
act as embedding medium. 7. Write about the infarction and shock?
• Tissue piece is placed on the disc/ metal Ans : Infraction : Definition - An area of ischemia
chuck with proper orientation. in an organ or tissue due to arterial insuffic-
ieicny or venous drainage block.
• The disc is placed on quick freezer shelf.
Cause : Thrombi or thrombo emboli
• Once the specimen is frozen, insert the disc
in the specimen head for sectioning. Types : Based on colour and presence pr
absence of bacterial contamination, there
• Hand wheel should be locked. are two types of infarcts.
• Knife or blade should be placed appropri- • Arterial-white infarct - Anaemic infarct.
ately.
ii. Shaft :Shaft is rounded in the upper half and triangular in the lower half.
Glomerular Filtration Rate is increased, when • Cations crosses the membrane through
extracellular fluid and blood volume are in- cotransport mechanism, while anions
creased. transfer through passive diffusion.
Renal Clearance : It is the volume of plasma • Urea and lipid soluble solutes gets diffused
from which the substance is completely re- passively across electrochemical gradient.
moved by both kidneys in unit time • Proteines are absorbed through endocytosis
Mass of the substance b. In Loop of Henle:
excreted in unit time • Water moves along with NaCl as a concu
Clearance of a substance =
Plasma concetration rrent mechanism.
of substance. • Na+ and chloride are the common ions
Urea Clearance: It is the volume of plasma reabsorbed at loop of henle.
from which urea is removed in one min. c. In Distal Convoluted Tubule:
B. TUBULAR REABSORPTION • Here, the reabsorption depends on the
The filtrate contains physiologically useful hormones.
substances namely glucose, amino acids, • Aldosterone promotes the excretion of K+
bicarbonates, phosphates, potassium and and reabsorption of Na+,Aldosterone
water which are absorbed in different parts mediated active transport cotransports H+,
of nephron. K+,Hco and C1‘ions and Antiduretic
Tubular Reabsorption occurs by two types: hormone activates the reabsorption of
water.
• Active Reabsorption: It is the movement of
molecules against the electrochemical gra- Collecting Duct: Urea in response to the
dient (up hill). This needs expenditure of concentration gradient and water in
energy which is derived from Adenosine response to ADH is absorbed
Triphosphate (ATP). C. TUBULAR SECRETION
* Passive Reabsorption: In this, molecules • In addition to reabsorption from renal tu-
move along the electrochemical (down hill) bules some substances are also secreted into
gradient. lumen from peritubular capillaries through
This process does not need energy. Sub- the tubular epithelial cells known as tubular
stances like chloride, urea and water are secretion or excretion.
reabsorbed passively The substances that are not required such
Site of Reabsorption: as foreign materials such as drugs (pencillin,
aspirin), which are not filtered from the skeleton embedded in it(sometimes also
blood are excreted from the body in the called bony pelvis or pelvic skeleton).
form of urine.
The pelvic region of the trunk includes the
H+, K+, creatinine, ammonium ion, marine bony pelvis, the pelvic cavity (the space
drugs moves from the blood through the enclosed by the bony pelvis), the pelvic floor
tubular wall into the filtrate. below the pelvic cavity, and the perineum
11. Write about human pelvis? below the pelvic floor.
Ans : Pelvic Girdle (Human Pelvis) : In human The pelvic skeleton is formed in the area of
anatomy, the pelvis (plural pelves) is either the back, by the sacrum and the coccyx and
the lower part of the trunk, between the anteriorly and to the left and right sides, by
abdomen and the thighs (sometimes also a pair of hip bones. The two hip bones
called pelvic region of the trunk), or the connect the spine with the lower limbs.
They are attached to the sacrum posteriorly, connected to each other anteriorly, and joined with
the two femurs at the hip joints.
The gap enclosed by the bony pelvis, called the pelvic cavity, is the section of the body underneath
the abdomen and mainly consists of the reproductive organs (sex organs) and the rectum.
Difference between male and female pelvis:
The female pelvis is adapted for pregnancy and child birth. It differs from male pelvis in the
following aspects:
• Bones are lighter and smoother.
• Body of pubis is broad and square.
• Pubic arch is wider.
• Pelvic cavity is broad and round.
• Sacrum is short and wide, coccyx is more movable.
Structure of the Hip Bone (or) Innominate age. The head of the femur articulates with the
bone: acetabulum to form the hip joint.
The hip bone is made up of the three parts: a. Ilium:
a. Ilium • The superior part of the hip bone is formed
by the ilium, the widest and largest of the
b. Ischium and
three parts.
c. Pubis.
The body of the ilium forms as the superior
Prior to puberty, the triradiate cartilage sepa- part of the acetabulum. Immediately above
rates these constituents. At the age of 15-17, the acetabulum, the ilium expands to form
the three parts begin to fuse. Their fusion forms the wing (or) ala.
a cup-shaped socket known as the acetabulum,
• The wing of the ilium has two surfaces. The
which becomes complete at 20- 25 years of
inner surface is concave, and known as the
iliac fossa, providing origin to the iliacus
Medical Lab Technician
MODEL PAPER -2 n 23