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May occur singly or in clumps. Mostly neutrophils, with granules n lobulation of nuclei. &lt;5 leucocytes/hpf are seen normally. Increased number of clumps are seen in cystitis, prostatitis and balanitis. &gt;30/hpf suggests acute infection. Eosinophils are increased in urine in tubulointerstitial disease, drug hypersesitivity. May occur singly or in clumps. Mostly neutrophils, with granules n lobulation of nuclei. &lt;5 leucocytes/hpf are seen normally. Increased number of clumps are seen in cystitis, prostatitis and balanitis. &gt;30/hpf suggests acute infection. Eosinophils are increased in urine in tubulointerstitial disease, drug hypersesitivity.
Under phase contrast, Trichomonas organisms appear much darker than the surrounding white blood cells.
This parasite is considered an important factor in the etiology of carcinoma of the bladder. The ova are elongated and are 60 X 160 microns. They are a yellowish color, slightly transparent and possess a delicate terminal spine.
Sperm may be present in urine sediment. Sperm have a characteristic oval body with a long thin tail and are 50 microns in length.
Oval fat bodies are degenerating tubular epithelial cells. They contain refractile fat droplets. These fats have been absorbed by the tubular cells after being leaked through abnormal glomeruli. They appear as grape-like clusters of variable size and are highly refractile.
Yeast can appear as single cells or in the budding form. In the budding form, yeast is easily identified as demonstrated on this slide. Yeast can be found in patients with cystitis due to yeast, usually candida, or as a vaginal contaminant from patient&apos;s with vaginal candidiasis.
Urine is formed in the kidneys, is a product
of ultrafiltration of plasma by the renal
Collection of urine
Early morning sample-qualitative
Random sample- routine
24hrs sample- quantitative
Post prandial sample-D.M
Catheterised- in infants, bedridden
Suprapubic needle aspiration
24 hour urine sample
1. For quantitative estimation of proteins
2. For estimation of vanillyl mandelic acid,
5-hydroxyindole acetic acid,
3. For detection of hormones in urine
4. For detection of microalbuminuria
Preservation of urine sample
HCl- for 24 hr urinary sample preservation
for adrenaline, noradr, VMA, steroids.
Toluene- as a physical barrier
Boric acid- general preservative
Thymol- inhibits bacteria, fungi
Formalin- for preservation of formed
For routine analysis, preservatives should
be avoided as they interfere with reagent
strip technique and chemical test for
Sample should be examined within 1-2 hrs
If delay is expected, sample can be kept in
refrigeration for max 8 hrs.
Reaction or urinary pH
Normal = 600-2000ml with night urine not in
excess of 400 ml.
Polyuria- >2000ml/24 hrs
Anuria-complete cessation of urine(<200ml)
Nocturia-excretion of urine by a adult of >500ml
with a specific gravity of <1.018 at night
(characteristic of chronic glomerulonephritis)
Color & appearance
Normal= clear & pale yellow due to the
presence of various pigments called
1. Colourless- dilution, diabetes mellitus,
diabetes insipidus, diuretics
2. Milky-purulent genitourinary tract
3. Orange- urobilinogen
4. Red-beetroot ingestion,haematuria,
5. Brown/ black- alkaptunuria, melanin
Urinary pH/ reaction
Reaction reflects ability of kidney to
maintain normal hydrogen ion
concentration in plasma & ECF
Tested by- 1.litmus paper
2. pH paper
3. Reagent strip method
Ketosis-diabetes, starvation, fever
UTI by E.coli
High protein diet
UTI by pseudomonas or Proteus
Normal= aromatic due to the volatile fatty
Ammonical – bacterial action(E. coli)
Fruity- ketonuria, starvation
Fishy- UTI with Proteus
Depends on the concentration of various
solutes in the urine.
Normal range- 1.003 to 1.035
- reagent strip method
- falling drop method
This method is based on the principle of
Take 2/3 of urinometer container with urine
Allow the urinometer to float into the urine
Read the graduation at the lowest level of
Correction of temperature & albumin is a
Urinometer is calibrated at 15or 200
So for every 3o
c increase/decrease add/subtract
All causes of oliguria
Gycosuria, DM, Dehydration, nephrotic
All causes of polyuria except gycosuria
DI, pyelonephritis, glomerulonephritis.
Fixed specific gravity (isosthenuria)=1.010
Seen in chronic renal disease when kidney
has lost the ability to concentrate or dilute
Normal adult with normal fluid intake wil
produce urine of 500-850 mOsm/kg water.
The normal kidney is able to produce
urine osmolality in the range of 800-1400
mOsm/kg water in dehydration and
minimal osmolality of 40-80 mOsm/kg
water during diuresis.
Tests for proteins
Test – HEAT & ACETIC ACID TEST
Principle-proteins are denatured & coagulated
on heating to give white cloud precipitate.
Method-take 2/3 of test tube with urine, heat only
the upper part keeping lower part as control.
Presence of phosphates, carbonates, proteins
gives a white cloud formation. Add 1-2 drops of
10% acetic acid, if the cloud persists it indicates
it is protein(acetic acid dissolves the
Other Tests for Protien
Nitric acid test
Sulphosalicylic acid test
Test with Esbach’s reagent
Protein % of Total Daily Maximum
Albumin 40% 60 mg
Tamm-Horsfall 40% 60 mg
Immunoglobulins 12% 24 mg
Secretory IgA 3% 6 mg
Other 5% 10 mg
TOTAL 100% 150 mg
Proteins in “Normal” UrineProteins in “Normal” Urine
Causes of proteinuria
Glomerular proteinuria: due to increased
permeability of glomerular capillary wall.
Selective( only albumin n transferrin bands seen)
Nonselective: pattern same as serum
e.g. nephrotic syndrome
Tubular proteinuria: in acute n chronic
pyelonephritis, heavy metal poisoning, TB
Overflow proteinuria: Bence jones
proteins(plasma cell dyscrasia),
hemglobin( intravascular hemlysis),
myoglobin(skeletal muscle trauma)
Hemodynamic proteinuria: seen in high
fever, hypertension, heavy exercise, CCF
Post-renal proteinuria: caused by
imflammatory or neoplastic conditions in
renal pelvis, ureter, bladder, prostate or
It is presence of albumin in urine above
normal level bt below detectable range of
conventional urine dipstick method.
Defined as urinary excretion of 30 to 300
mg/24 hrs of albumin in urine.
Significance of microalbuminuria
an indicator of subclinical cardiovascular
an important prognostic marker for kidney
in diabetes mellitus (earliest sign of renal
damage in DM)
increasing microalbuminuria during the first 48
hours after admission to an intensive care unit
predicts elevated risk for acute respiratory failure
, multiple organ failure , and overall mortality
Detection of microalbuminuria:
Methods for detection :
Measurement of albumin creatinine ratio
in random urine sample.
Measurement of albumin in early morning
Measurement of urine in 24 hr sample.
Bence Jones proteins
These are monoclonal immunoglobulin light
chains (kappa or lamda) synthesized by
neoplastic plasma cells.
seen in multiple myeloma, macroglobulinemias,
Test- Thermal method(waterbath):
Proteins have unusual property of
precipitating at 400
c & then dissolving
when the urine is brought to boiling(1000
reappears when the urine is cooled.
Test for sugar Test-BENEDICT’S TEST(semiquantitative)
Principle-benedict’s reagent contains
cuso4.In the presence of reducing sugars
cupric ions are converted to cuprous oxide
which is hastened by heating, to give the
Method- take 5ml of benedict’s reagent in a
test tube, add 8drops of urine. Boil the
Detects all reducing substances like
glucose, fructose, & other reducing
Sensitivity of the test is about 200 mg
reducing substance per dl of urine.
To confirm it is glucose, dipsticks can be
used (glucose oxidase)
Reagent strip method:
Specific for glucose.
Based on glucose oxidase peroxidase
More sensitive (sensitivity- 100 mg
Glu + oxygen---- gluconic
oxidised chromogen(blue)+ H2O
Other Methods of detecting glycosuria
Causes of glycosuria
Glycosuria with hyperglycaemia-
diabetes,acromegaly, cushing’s disease,
hyperthyroidism, drugs like corticosteroids.
Glycosuria without hyperglycaemia-
renal tubular dysfunction
β-hydroxy butyric acid
They are products of fat metabolism
Principle-acetone & acetoacetic acid react
with sodium nitroprusside in the presence
of alkali to produce purple colour.
Method- take 5ml of urine in a test tube &
saturate it with ammonium sulphate. Then
add one crystal of sodium nitroprusside.
Then slowly run the liquor ammonia along
the sides of the test tube.
Formation of purple coloured ring at
junction indicates + test
Acetest tablet test
Ferric chloride test(Gerhardt’s test)
Reagent strip method
Hart’s test for beta hydroxy butyric acid
Causes of ketonuria
Non-diabetic causes- high fever,
starvation, severe vomiting/diarrhoea
Glycogen storage diseases
Test- fouchet’s test.
In 5 ml of urine add 2.5 ml of 10% Barium
chloride and mix well. Then filter to obtain
precipitate. To ppt add 1 drop of fouchet’s
reagent. Development of blue green colour
indicate + test.
Principle: Bilirubin Adsorbs to the Barium Chloride
and results in green color formation when
fouchet’s reagent is added.
Obstruction to biliary tract
Gmelin’s test (nitric acid)
Lugol’s iodine test
Reagent strip with diazo reagent
Test- ehrlich test
In 5 ml of urine add 0.5 ml of Ehrlich’s
reagent(HCl 20 ml, d/w 80 ml, p-
dimethylaminobenzaldehyde 2 gm). Allow
to stand for 5 min. development of pink
colour indicates +test.
Causes-hemolytic jaundice, early
hepatitis, hepatocellular jaundice.
Hay’s sulphur test:
In 5 ml of urine sprinkle a pinch of
sulphur particles. If bile salt is present
sulphur particles will sink to the bottom
because bile salts lowers the surface
tension of urine.
Blood in urine
Test- BENZIDINE TEST
Principle-The peroxidase activity of hemoglobin
decomposes hydrogen peroxide releasing
nascent oxygen which in turn oxidizes benzidine
to give blue color.
Method- mix 2ml of benzidine solution with 2ml
of hydrogen peroxide in a test tube. Take 2ml of
urine & add 2ml of above mixture. A blue or
green color within 5 min indicates + reaction.
Causes of hematuria
Pre renal- bleeding diathesis,
Renal- trauma, calculi, acute & chronic
glomerulonephritis, renal TB, renal
Post renal – severe UTI, calculi,
trauma, tumors of urinary tract
Type Plasma color Urine color
Hematuria normal Smoky red
Myoglobunuria Pink, normal
A well mixed sample of urine (12 ml) is
centrifuged in machine for 5 min at 1500 rpm.
The top liquid part (the supernatant) is
discarded. A drop of urine left at the bottom of
the test tube (the urine sediment) is placed on
glass slide and covered with cover slip. It is
examined under high power.
Contents of normal urine m/s
Contains few epithelial cells, occasional
RBC’s, few crystals.
Urinary casts are cylindrical aggregations
of particles that form in the distal nephron,
dislodge, and pass into the urine. In
urinalysis they indicate kidney disease.
They form via precipitation of
Tamm-Horsfall mucoprotein which is
secreted by renal tubule cells.
The most common type of cast, hyaline
casts are solidified Tamm-Horsfall
mucoprotein secreted from the tubular
Seen in fever, strenuous exercise,
damage to the glomerular capillary
Granular casts can result either from the
breakdown of cellular casts or the
inclusion of aggregates of plasma proteins
(e.g., albumin) or immunoglobulin light
indicative of chronic renal disease
Formed by the breakdown of lipid-rich
epithelial cells, these are hyaline casts
with fat globule inclusions
They can be present in various disorders,
diabetic or lupus nephropathy,
Acute tubular necrosis
Formed by the adhesion of metabolic
breakdown products or drug pigments
Pigments include those produced
endogenously, such as
hemoglobin in hemolytic anemia,
myoglobin in rhabdomyolysis, and
bilirubin in liver disease.
Though crystallized urinary solutes, such
as oxalates, urates, or sulfonamides, may
become enmeshed within a hyaline cast
during its formation.
The clinical significance of this occurrence
is not felt to be great.
Red cell casts
The presence of red blood cells within the
cast is always pathologic, and is strongly
indicative of glomerular damage.
They are usually associated with nephritic
This cast is formed by inclusion or
adhesion of desquamated epithelial cells
of the tubule lining.
These can be seen in
acute tubular necrosis and
toxic ingestion, such as from mercury,
diethylene glycol, or salicylate.
Urine dipstick is a narrow plastic strip which
has several squares of different colors
attached to it. Each small square represents
a component of the test used to interpret
urinalysis. The entire strip is dipped in the
urine sample and color changes in each
square are noted. The color change takes
place within 2 minutes from dipping the
strip. If read too early or too long after the
strip is dipped, the results may not be
Chemical AnalysisChemical Analysis
Urine DipstickUrine Dipstick
The squares on the dipstick represent the
following components in the urine:
∀ specific gravity (concentration of urine),
∀ acidity of the urine (pH),
∀ protein in the urine (mainly albumin),
∀ glucose (sugar),
∀ bilirubin and
The main advantage of dipsticks is that
2. easy to interpret,
3. and cost-effective
The main disadvantage is that the
1. Information may not be very accurate as
the test is time-sensitive.
2. It also provides limited information about
the urine as it is qualitative test and not a
quantitative test (for example, it does not
give a precise measure of the quantity of
Automated urine analyser:
It is based on the principle of flow
This system classifies particles based on
fluoroscent intensity ( cells are stained
with 2 fluoroscent dyes), electrical
impedance and forward angle light scatter.
Next generation automated image based
urinalysis system iQ200 is a walk away
This system uses flow imaging analysis
technology and Auto Particle Recognition
It requires minimum of 3 ml of urine and
quantitates particles in 2 microlitre of
Can analyse 60 urine samples per hr.
Henry’s Clinical diagnosis and
Management of Laboratory Methods
Clinical Pathology Kawthalkar
Clinical Pathology Sabitri Sanyal
Basics of Body fluids by Akhil Bansal