Erbil Polytechnic University

Shaqlawa technical Institute

MLT department

Lab.
technology

Ali Zainal
MSc in physiology
[email protected]
 Urine analysis
Urinalysis is a group of tests performed most
frequently on random specimen. It is one of the
most helpful indicators of health and disease,
especially, it is useful as a screening test for the
detection of various endocrine or metabolic
abnormalities in which the kidneys function
properly but they will excrete abnormal
amounts of metabolic end-products specific of a
particular disease
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 Physiology of the Kidney and Formation of Urine

Approximately 1,200 milliliters of blood flow

through the kidneys each minute. This
represents about one-fourth of the total blood
volume. The blood enters the glomerulus of
each nephron by passing through the afferent
arteriole into the glomerular capillaries. The
capillary walls in the glomerulus are highly
permeable to water and the low molecular-
weight components of the plasma.
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They filter through the capillary walls and the
closely adhering membrane of Bowman's
Capsule into Bowman's Space from where the
plasma ultra-filtrate passes into the tubule
where reabsorption of some substances,
secretion of others, and the concentration of
urine occur. Many components of the plasma
filtrate, such as: glucose, water, and amino acids,
are partially or completely reabsorbed by the
capillaries surrounding the proximal tubules.

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In the distal tubules, more water is reabsorbed and
potassium and hydrogen ions are secreted. The Loop of
Henle and the system of collecting tubules are the
principal sites where the urine is concentrated as a
mechanism for conserving body water. Urine formed by
the three physiological processes that are by
glomerular filtration, tubular reabsorption, and
tubular secretion, is collected by the collecting duct
and passes into bladder through ureters and then
comes out through urethra.

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 Collection of Urine Specimen
• In order to make Urine analysis reliable the
urine must be properly collected. Improper
collection may invalidate the results of the
laboratory procedures.

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 Urine Containers
• There are many types of containers used for
collecting urine. Before specimens are
collected, the containers must be cleaned and
thoroughly dried. Disposable containers of
plastic or coated paper are available in many
sizes and are provided with lids to reduce
bacterial and other types of contamination.

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Urine cup

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Special polyethlene bags are available for
collection of urine from infants and children
who are not toilet trained.

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 Types of Specimen

• First Morning Specimen :

• a specimen obtained during the first urination of the
day.
• Most concentrated
• Bladder incubated
• Best for:
• Nitrite
• Protein
• Microscopic examination

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• Random Specimen :
• a specimen obtained at any time during examination.
• Most convenient
• Most common
• best for:
• Chemical Screen
• Microscopic examination

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• Second-voided Specimen
• In this case first morning specimen is
discarded and the second specimen is collected
and tested. Such type of specimen is good for:
• Reflection of blood glucose.
• Keeping of formed elements intact

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• Postprandial
• a specimen obtained 2 hours after meal.
• for glucose.

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• 24- Hour specimen
• a specimen obtained within 24 hours.
• Necessary for quantitative tests, especially for
quantitative determination of protein and
GFR.

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• Mid- stream Specimen
• a specimen obtained from the middle part of
the first urine.
• It is commonly used for routine urinalysis.
• It is also important for bacteriological urine
culture.

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 Preservation of Urine Specimen

• Urine should be examined immediately, because some urinary

components are unstable.
Long standing of urine at room temperature can cause:
• Growth of bacteria
• Break down of urea to ammonia by bacteria leading to an
increase in the pH of the urine and this may cause the
precipitation of calcium and phosphates.
• Oxidation of urobilingen to urobilin.
• Destruction of glucose by bacteria.
• Lysis of RBCs, WBCs and casts

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 Type of Examination in Routine Urinalysis
• 1-Physical Examination of Urine
• Volume: Normally, 600 – 2000 ml of urine is voided per 24 hr.
• Color: morning it has dark yellow color, while in the afternoon or evening,
the color ranges from light yellow to colorless. Normal urine color varies
from straw (light yellow color) to dark amber (dark yellow).
• Odor: has faint aromatic odor, which comes from volatile acids, normally
found in urine, mostly, ammonia.
• Appearance, Degree of cloudiness of the urine is described by using
common terms, starting by clear to turbid i.e. clear, hazy, cloudy, very
cloudy and turbid.
• pH : Normally, freshly voided urine pH range from 5-7 in healthy
individuals, and average is pH 6.
• Specific gravity: ratio of the weight of a fixed volume of solution to that of
the same volume of water at a specified temperature 20o C

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 2-Chemical Examination of Urine

• Glucose: either physiological (large ingestion of

carbohydrates, pregnancy, Renal Glycosuria) or
pathological (Diabetes mellitus, Hyperthyroidism,
Hyperadrenalism, Hyperpitutarism)

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Protein Condition (s)
Strenuous Physical Exercise
Albumin Emotional Stress
Pregnancy
Infections
Glomerulonephritis
Newborns ( First Week )
Glomerulonephritis
Globulin Tubular Dysfunction
Hematuria
Hemoglobin Hemoglobinuria
Fibrinogen Severe Renal Disease
WBCs in Urine
Nucleoprotein Epithelial Cells in Urine
Multiple Myeloma
Bence jones Leukemia
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• Ketones
• Ketone bodies: They are normally not
detectable in the blood or urine are a group of
three related substances such as, acetone,
acetoacetate and β -hydroxybutyrate. Ketone
bodies are normal products of fat
metabolism..

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• Bilirubin (normal range 0.02 mg/dl)
• Bilirubin is a waste product that must be
eliminated from the body. It is formed by the
breakdown of hemoglobin in the reticulo
endothelial cells of the spleen and bone
marrow, and then transported to the liver.

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• urobilinogen (Normally 1-4 mg/day)
• In the intestine, most of the bilirubin is
converted to urobilinogen or stercobilinogen
by the action of certain bacteria that make up
the intestinal flora. Urine is often tested for
increases in urobilinogen when investigating
hemolytic jaundice or liver disorder in which
liver function is impaired.

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• Blood (normal 1.0 - 1.4 g/1.)
• Hemoglobin appears in the urine when there
is extensive or rapid destruction (hemolysis) of
circulating erythrocytes that the
reticuloendothelial system cannot metabolize
or store the excessive amounts of free
hemoglobin.

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• Nitrite
• The detection of nitrite in the urine can be
used to indicate the presence of bacteria such
as Escherchi coli, proteius, klebsiella,
enterobacter, citrobacter, and salmonella will
reduce urinary nitrate to nitrite

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• Leukocyte Esterase
• The presence of leukocytes indicates
inflammation at some point along the
urogenital tract

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• Melanin
• Melanin occurs in metabolic tambours
especially with metastasis of liver.

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 Microscopic Examination of Urine

• Red Blood Cells:

• Normally RBCs appear in the fresh sample as intact,
small and faint yellowish discs, darker at the edges,
Measure 7-8 μm in diameter. When the number of
RBCs is found more than their normal range, usually
greater than 5 RBCs/HPF it may indicate Acute and
chronic glomerulonephritis, Renal stone, Cystitis,
Prostates, Trauma of the kidney and Presence of
disease conditions in the urinary tract

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• White Blood Cells:
• normally, clear granular disc shaped ranged between
0-4 WBC/HPF, Measure 10-15 μm, the nuclei may be
visible. increased number of leukocyte urine are seen
in case of : Urinary tract infection, All renal disease,
Bladder tumor, Cystitis, Prostates, Acute or chronic
bacterial infection

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• Epithelial cells:
• Normally few epithelial cells (0-2 / HPF) can be found, Their
size differs depending on the site from which they originated.
Presence of epithelial cells in large number, mostly renal types
may indicate:
• Acute tubular damage, Acute glomerulonephritis and Silicate
over dose, also the presence of large number of epithelial
cells with large number of Leukocytes and mucus trades
(filaments) may indicate Urinary Tract Infections (UTI).
• Epithelial cells distribution reported after looking under 10x
(low power objective) of the microscope

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• Usually they are reported semi quantitatively by saying
• Occasional epithelial cells /LPF ……1-3 epithelial cells seen in
the whole LPF
• Few epithelial cells / LPF................ 2-4 epithelial / LPF
• Moderate epithelial cells / LPF......... 6-14 epithelial / LPF
• Many epithelial cells / LPF......... 15-25 epithelial/ LPF
• Full of epithelial cells / LPF..............……...
when the whole field of 10 x objective covered by epithelial cells.

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• Casts:
• Formed by precipitation of proteins, and
aggregation of cells within the renal tubules.
Most of them dissociate in alkaline urine, and
diluted urine (specific gravity ≤ 1.010) even in
the presence of proteinurea

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• a. Hyaline Casts
• Normal range: 0-2/HPF produce as a result of
possible damage of glomerular capillary membrane
• b. Grandular Casts
• c. waxy casts: Not seen in the normal individuals
• d. fatty casts: Not seen in the normal individuals
• e. cellular casts: Not seen in the normal individuals

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 Bacteria
• Normally, bacteria are not seen in the healthy individual’s
urine
• To check the presence or absence of bacteria a technician can
either check for Nitrate that was formed in the urine after
breakdown of nitrite into nitrate by the metabolic action of
bacteria. Also, presence of pus cells within the drop of urine
or its sediment.
• Further the observed bacterial cell can be identified by
bacteriological culture. Presence of bacteria may indicate the
presence of UTI

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 Yeasts
• Yeast cells are fungi that are not normally seen
in health individuals, Oval in shape, and
usually form budding. have high refractive
index. They are usually of candida species
(Candida albicans) and are common in
patients with UTI, vaginitis and Diabetic
mellitus

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 Parasites
• Parasites that can be seen in urine microscopy
are:
• Trichomonas vaginalis
• Schistosoma haematobium
• Wuchereria bancroftie
• and other parasites also may occur due to
contamination

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 Crystals
• Mostly occur during metabolic abnormalities
and excessive consumption of certain
foodstuffs. May be classified into acidic, basic,
and both acidic and basic

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 I. Acidic Urine Crystals
• a-Amorphous Urates:
• Normally present in urine in different quantity,
have pink to “brick red” color.

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• b-Uric Acid Crystals:
• Polymorphs (different in shape) i.e. square,
prism, hexagonal, rostelles, yellow to yellow
brown in colorm. Size is 30-150 μm (gout and
leukemia )

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• c-Cystine Crystals: rarely found (Wilson disease and
pyelonephritis )

• d-Cholesterol: rarely found (elevated cholesterol

chyluria )

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• e-Tyrosin : found rare (protein break down problem, or severe
liver disease

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 II. Alkaline Urine Crystals
• a. Amorphous Phosphates: Normally seen in alkaline urine.
Small, whitish granules usually seen scattered

• b. Calcium Carbonate: Less commonly seen.Colorless, have

needle, spherical or dumbbells shape.

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• c. Calcium Phosphates: Seen in small amount in normal individual
urine, and when they are in large amount, may indicate chronic
cystitis, or prosthetic hypertrophy, have star or needle shape.

• d. Ammonium Bruits (Urates): Normally seen in alkaline urine,

have bundle of needles or “thorn apple” sphere shape.

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 III. Alkaline, Neutral, or Slight Acidic Urine
Crystals
• Triple Phosphates : Colorless and refractive, have
“coffin lids” 3 to 4 to 6 – sided prism, fern leaf or
star shape

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IV .Acidic, Neutral, or Basic Urine Crystals

• Calcium Oxalate Crystal: Are colorless and refractive,

have octahedral, envelope, shape., size 10-12 μm.

• Normally seen in small amount, after consumption of

high calcium, or oxalate rich foods, such as milk,
tomatoes, asparagus, and orange

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 Artifacts
• -Spermatozoa: it frequently seen n the urine of male and they
may be seen in the urine of female when the urine collected
after coitus, usually do not report it.
• -Mucus Thresds: Formed by the precipitation of mucoprotein
in cooled urine. Normally little mucus thread seen in normal
individuals, have fine, fiber like appearance, may indicate UTI.
• -Starch granules
• -Fat droplets
• -Oil droplets

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Urine Dipstick examination

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