CLINICAL MICROSCOPY (LECTURE)

GARCIA

#TeamCSPage 1

Lesson 1: RENAL ANATOMY &

PHYSIOLOGY
*Kidneys major organ in urinary system
*Urethra difference in male and female
(length)

NEPHRON
1.3M per kidney
Responsible for the filtration of
blood and formation of urine
Basic unit of the kidney

FUNCTIONS OF THE KIDNEYS

Removal of metabolic waste
products (major function).
- Wastes: urea and creatinine
Regulation of water & electrolytes.
- Kidneys can filter 180L of
blood/day and 1.2L of urine per
day
Maintenance of the bodys acidbase balance.
- Blood pH = 7.4 (7.35 7.45)
- Maintenance of blood pH
through: BUFFER, PULMONARY
& KIDNEYS
**Kindeys decide which one to retain and
which one to excrete.

A. Renal Corpuscles
1. Glomerulus
2. Bowmans Capsule

Responsible
for the
filtration of
urine

B. Renal Tubules

Responsib
le for the
1. Proximal Convuluted Tubules (PCT)
reabsorpti
2. Loop of Henle (LOH)
on and
secretion

3. Distal Convuluted Tubule (DCT)

PASSAGE OF BLOOD

PARTS OF THE KIDNEY

**Afferent arteriole -> Glomerulus ->

**If particles are big, Glomerulus ->
efferent arteriole
**If particles are small, Glomerulus ->
Bowman's Capsule
Glomerulus responsible for
filtration and granular appearance
of cortex
Cortex outer portion of the
kidney
Medulla inner part of the kidney
Calyx bigger tubules
Renal Pelvis compiled calyxes

** Ultrafiltrate of plasma same

composition with plasma but does not
involve blood cells

FILTRATION BARRIER
1. Endothelial Cells
2. Basement Membrane

3. Podocytes

PROCESS OF URINE FORMATION

Regulation of blood volume,

blood pressure & Na level
(stimuli that activate Renin).

**Angiotensinogen free blood protein in

the plasma

1. Glomerular Filtration
2. Tubular Reabsorption
3. Tubular Secretion

2 TYPES OF TRANSPORT
1. Active Transport
2. Passive Transport
LOCATION

PCT

MODE OF
TRANSMISSI
ON

Passive
Active

LOH

Passive

DCT

Active

SUBSTANC
E
Water, Cl, K,
Urea, Na,
HCO3,
glucose,
amino acid,
proteins, PO4,
SO4, Mg, Ca
& Uric Acid

Water,
Urea, Na,
Cl
Na, Cl, SO4,
Uric Acid

Collecting
Passive
Water, Cl
Tubule
Active
Na
(Cortical)
Collecting
Tubule
Water,
Passive
(Medullary
Urea
)
**Water is always reabsorbed
Normall
y found

REGULATION OF KIDNEY FUNCTION

Juxtaglomerular Apparatus:
Renin
- Macula densa (crowded cells in
the DCT and LOH)
- Juxtaglomerular cells (afferent
arteriole)

Maximal Reabsorptive Capacity

- Renal Treshold (for glucose 160
180 mg/dL; normal value of
glucose is 100 mg/dL)
- Countercurrent mechanisms
between the LOH & vasa recta
- ADH/Arginine vasopressin
*How much blood is filtered by the
glomerulus? 1.25mL/min
*How much liters of blood is filtered
by day? 180L
*How much urine is produced? 600
1,800mL (1,200mL average)

URINE COMPOSITION
94% water
Urea (majority), Cl, Na, K
Phosphates, sulfate, creatinine and
uric acid

RENAL FUNCTION TESTS

A. Concentration (amount of solute)
Osmolality (most preferred)
- osmoies of solute particles per
kg of solvent

Good indicator of the kidneys

function to concentrate urine
- 800mOsm/kg
Specific Gravity
- Density of urine to the density
of pure water
- < 1.025

Consistency in the plasma level

Substance availability in the body
Availability of the test to chemical
analysis

CREATININE

Values depend on:

~degree of hydration
~physical activities
~stress

Some creatinine is secreted by the

tubules and secretion increases as
blood level rises
Chromogens present in human
plasma react in the chemical
analysis
Bacteria break down urinary
creatinine
Diet affects results

B. Fluid Deprivation Test

ADH secretion problems
C. Renal Clearance/Clearance Test/
Glomerular Filtration Test
How long or how fast the kidney
performs

D. Tubular Secretion and Renal Blood

Flow

Correction Factor = 1.73m2

- Average body surface area
- Average muscle mass

p-aminohippuric acid (PAH) Test

Phenolsulfonphthalein (PSP)
Titrable acidity & Urinary ammonia

SUBSTANCES USED FOR CLEARANCE

TEST
Urea (from CHON) traditionally
used
Creatinine (from muscle
metabolites) replaced urea for
common test; ideal but not
preferred
Inulin
Beta macroglobulin
Radioisotopes
Cystatin C

CLEARANCE TEST
Glomerular Filtration Test
**SUBSTANCE MUST BE:
Neither reabsorb nor secreted by
the tubules
Stability in the urine during the
24hr collection

HISTORY

Hieroglyphics

Uroscopy (Hippocrates) - study of urine

Ant and taste testing (Sasruta)

Urine boiling (Frederick Dekkers) - due to

- inexpensive

presence of albumin

- fountain of information

Addis Count (Thomas Addis) - micrscopic;

discovered analytes

DEFINITION

WHY STUDY URINE?

- by CSLI, testing urine with pro... commonly

performed in an expeditious, reliable, accurate,

- Relatively readily collected and easily available

safe and cost-effective manner

specimen

- less invasive
IMPORTANCE

- contains information about the body's major

metabolic functions

- fluid biopsy of kidney

- aid in the diagnosis of disease (ex UTI)

- help screen asymptomatic, congentital or

hereditary diseases

- monitor disease progression

FIRST MORNING SPECIMEN

- monitor therapy effectiveness and complications

- collected after you wake up (should have empty
bladder before going to sleep)

SPECIMEN TYPES

RANDOM URINE SPECIMEN

- collected any time of the day

- for routine urinalysis and drug testing

- routine screening tests

- must be used within 2hrs after collection

- highly concentrated

- pregnancy testing (excrete hcg)

- orthostatic proteinuria (due to posture)

Random

- Syrcardian Rhythm

1st
Morning

- urobilinogen: 2-4pm peak excretion

Ortho

- should always start and end with an empty

bladder

True pro

- conc of substance to be measured changes

with diurnal variations and daily activities

TIMED SPECIMEN

FASTING/SECOND MORNING

- Second voided urine

- for quantitative test

- determine amount of substance in urine

- 12hr, 24hr (most common), 2hpp

2 HOUR POST PRANDIAL

- 2 hours after eating

GLUCOSE TOLERANCE SPECIMEN

- routine urinalysis vs urine culture

MIDSTREAM CLEAN CATCH

- wash the genitalia

COLLECTION TECHNIQUES

- first voided urine, discard

- biohazardous substance
- safer, less traumatic
- clean dry container, disposable, wide mouthed
clear material

- labelling and requisition form is a must

SUPRAPUBIC ASPIRATION

- sterile needle to bladder of individual

- catheter

- for bacterial and cytologic testing

- bacterial culture

- collected under sterile conditions

PEDIATRIC COLLECTION

3-GLASS COLLECTION

- pediatric bags with adhesives

- almost similar to midstream clean catch
- check every 15-20 minutes
- for prostatic infection (presence of WBC)

- 1st: voided

CATHETERIZED SPECIMEN

- 2nd: midstream; control for presence of UTI

- 3rd: infection

- containers with bleach & detergent residue

- improper or non-matching labeling

- Quantity not sufficient (QNS)

DRUG SPECIMEN COLLECTION

- Urine contaminated with feces

- chain of custody be trace if there's tampering

- well supervised

- Inadequently checked containers

- Urine squeezed from diapers

- check for urea and creatinine content

URINE PRESERVATION

- Not more than 2 hours

UNSUITABLE SPECIMEN

- Add chemical preservatives/refrigerate if delayed

CO2

- No PERFECT preservative

Bilirubin

Decrease

Photo oxidation

Glucose

Decrease

Glycolysis &
consumption of

- Each preservative has disadvantages and

advantages, MT should decide what best to use

cells
Ketones

Decrease

Evaporation

Nitrite

Increase

Conversion of
nitrate to nitrite

Urobilinogen

Decrease

Oxidation to
urobilin

CHANGES THAT OCCUR IN IMPROPERLY

PRESERVED SPECIMEN

Crystals

Increase

Crystallization

Cells &

Decrease

Disintegration

Bacteria

Increase

multiplication

Trichomonas

Decreased

Loss of motility,

Casts

death
Color

Darker

Oxidation or
reduction of
metabolites

Clarity

Odor

pH

Turbid/

Bacterial

Decreased

proliferation

Ammoniacal/

Conversion of

Increased

urea to ammonia

Increase,

Breakdown of

alkaline

urea to ammonia
by ureaseproducing
bacteria/loss of

Urine
ultrafiltrate of plasma
urea + other organic +
inorganic substance
95% water, 5% solutes
Factors dietary intake, physical
activity, body metabolism, endocrine
functions
Urea
produced in the liver
breakdown of CHON and amino
acids
50% of total dissolved solids
Creatinineproduct of creatinine
metabolism by muscles

 Uric Acid- product of nucleic acid

breakdown in food and cells
Chloride- major inorganic solid (followed
by Na, K)
found in combination with
sodium and other inorganic compounds
Sodium primarily from salt
Potassiumcombined with salts
Phosphatecombines with Na (to
buffer blood)
Ammoniumregulates blood and
tissue fluid acidity
Calcium binds with chloride,
sulfate, and phosphate
Urine Volume

Oliguria

Anuria

Nocturia
urine
Polyuria

primarily water
Determined by the
amount of hydration
- Factors: fluid intake,
fluid loss, secretion of
ADH, need to excrete
increased amounts of
dissolved solids
- Normal urine daily
output: 1200mL to
1500mL ( 600mL to
2000mL)
urine output
infants: < 1mL/kg/hr
children: < 0.5mL/kg/hr
adults: < 400mL/day
dehydration, excessive
water loss
cessation of urine flow
kidney damage or flow
of blood
nocturnal excretion of
-

urine daily volume

children: > 2.5 to

3mL/kg/day
adults: > 2.5 L/day
diabetes mellitus /
insipidus, diuretics
Diabetes Mellitus- defect in pancreatic
production or function of insulin = glucose
conc appears diluted = SG
Diabetes Insipidus- production / function of
ADH = dilute, SG

Specimen Rejection1. in unlabelled

containers
2. Non-matching labels
and requisition forms
3. Contaminated with
feces
4. Containers with
contaminated exteriors
5. Insufficient quantity
6. Improperly
transported
Specimen Preservation
Common: refrigeration
o 2-8 C
o bacterial growth and
metabolism

PRESERVATIVES

ADVANTAGES

DISADVANTAGES

ADDITIONAL
INFO

Refrigeration

Does not interfere

Precipitates

Prevents bacterial

with chemical tests

amorphous

growth for 24 hrs

phosphates &
ureases
Boric acid

Prevents bacterial

Interferes with drug

Keeps pH at about

growth and

& hormone analyses

6.0; can be used for

metabolism

urine culture
transport

Formanlin

Excellent sediment

Acts as a reducing

Rinse specimen

(formaldehyde)

preservative

agent, interfering

container with

with chemical tests

formalin to preserve

for glucose, blood,

cells and casts

LE & Cu reduction
Sodium fluoride

Drug analyses

Inhibits rgt strip tests

for glu, blood, &
leukocytes

Commercial

When refrigeration

Check tablet

preservative tablets

is not possible; have

composition to

controlled conc. to

determine possible

min. interference

effects on desired
tests

Urine Collection

Contains collection

Kits (Becton,

cup, transfer straw,

Dickinson,

C&S preservative

Rutherford, NJ)

tube or UA tube

Light gray and gray

Sample stable at RT

Do not use if urine is

Preservative is boric

C&S tube

for 48 hrs; prevents

below minimum fill

acid, sodium borate

bacterial growth and

line

& sodium formate

Use on automated

Must refrigerate

Round or conical

instruments

w/in 2 hrs

bottom, no

metabolism
Yellow UA Plus tube

preservative
Cherry red/yellow

Stable for 72 hrs at

Must be filled to min

Preservative is

Preservative Plus

RT; instrument-

fill line.

sodium propionate,

tube

compatible

Bilirubin and

ethyl paraben, and

urobilinogen may be

chlorhexidine; round

decreased if

or conical bottoms

specimen is exposed
to light and left at
RT

PHYSICAL EXAMINATION OF URINE

Color - varies from colorless to black

Normal Urine Color

Red/ Pink/ Brown depends on the pH of

the urine

Long standing Brown Acidic Urine

due to oxidation of hemoglobin to
methemoglobin

Fresh Brown Urine containing blood

glomerular bleeding resulting from the
conversion
of
hemoglobin
to
methemoglobin

Pale yellow to dark yellow

Yellow color is caused by urochrome

(Thudichum, 1864)

Hemoglobin and myoglobin produces

red urine

Urochrome product of endogenous

metabolism

If specimen is red and clear, hemoglobin

or myoglobin is present

Uroerythrin and Urobilin present in

much smaller quantities

Uroerythrin pink pigment resulting in

the precipitation of amorphous urates

Urobilin

oxidation
product
urobilinogen, orange-brown color

of

Nonpathogenic causes:

Dark Yellow/ Amber/ Orange

Caused by the presence of abnormal

pigment bilirubin

Presence of bilirubin could be suspected if

yellow foam appears when specimen is
shaken
Large amount of white foam could
indicate presence of large amount of
protein

Urine specimen that has bilirubin may also

contain hepatitis virus

Photo-oxidation of bilirubin imparts

yellow-to green color caused by biliverdin

Yellow-orange specimen caused

phenazopyridine or ago-gantrisin
people who have UTI

by
to

Red/ Pink/ Brown

Myoglobinuria - breakdown of skeletal muscles,

more reddish-brown in color
Urine specimens containing porphyrins may also
appear red, resulting from the oxidation of
porphobilinogen to porphyrins

Abnormal Urine Color

Hemoglobinuria - in vivo breakdown of RBCs

accompanied by red plasma

Mostly caused by the presence of RBCs

menstrual contamination

ingestion of highly pigmented foods and

medications

Brown/ Black

Presence of melanoma or homogenistic

acid

Homogenistic acid metabolite of

phenylalanine; persons with alkaptonuria

Blue/ Green

Bacterial/ Urinary Tract Infections by

Pseudomonas species

Intestinal infections caused by urinary

indicant

Infections caused
providencia species

by

Klebsiella

or

Clarity - Refers to the transparency or turbidity of

the urine specimen; Visual examination
Normal Clarity

Usually clear

Precipitation of amorphous phosphates

and carbonates may cause white
cloudiness

Odor

Nonpathologic Turbidity

Squamous Epithelial Cells, Mucus

Amorphous
urates

Semen, spermatozoa

Fecal contamination

Radiographic contrast media

Talcum powder

Vaginal creams

phosphates,

Reagent Strip SG based on the change in the

pKa of a polyelectrolyte in an alkaline medium,
high concentration, more H ion, lower pH

carbonates,

Pathologic

Odor
Arom
atic

Cause
Normal

Fruity,
sweet
Mous
y
Sweat
y feet

Ketones
(DM,
starvation,
vomiting)
Phenylketo
nuria
Isovaleric
anemia

Bleac
h

Contaminat
ion

Odor
Foul,
ammo
nia-like
Maple
Syrup

Cause
Bacterial
decomposi
tion, UTI
Maple
Syrup
Urine Dse

Rancid

Tyrosinemi
a
Methionine
malabsorp
tion

Cabba
ge

RBCs, WBCs

Bacteria, Yeast

CHEMICAL EXAMINATION OF URINE

Non squamous epithelial cells

REAGENT STRIP

Abnormal crystals

Lymph fluid

Lipids

Specific Gravity

Density of the solution compared with the

density of a similar volume of distilled
water at a similar temperature

Isosthenuric SG 1.010
Hypersthenuric SG above
Hyposthenuric SG below

Provides a simple, rapid means for

performing medically significant chemical
analysis of urine, including pH, protein,
glucose, ketones, blood, bilirubin,
urobilinogen, nitrite, leukocytes, and
specific gravity.
Two major types of reagent stip:
Multistix
Chemstrip
ERROR CAUSE BY IMPROPER TECHNIQUE

1. Unmixed specimen

Random specimens - fall at 1.015 to 1.030

2. Allowing the strip to remain in the urine for an

extended period

Refractometry concentration of dissolved

particles in a specimen by measuring its refractive
index

3. Excess urine remaining on the strip

Osmolarity affected only by the number of

particles present, determined by measuring a
property that is mathematically related to the
number of particles in the solution

4. The timing for reactions to take place varies

between tests and manufacturers
5. A good light source
6. The strip must be held close to the color chart
without actually being placed on the chart

7. Reagents strips and color charts from different

manufacturers are not interchangeable
8. Specimens that have been refrigerated must
be allowed to return to RT prior to reagent testing
HANDLING AND STORING REAGENT STRIPS

Packaged in opaque containers with a

desiccant to protect them from light and
moisture
Reagent strips be stored at room
temperature below 30oC
QUALITY CONTROL OF REAGENT STRIPS

Must be checked with both positive and

negative controls minimum of once every
24 hours
Distilled water is not allowed as negative
control because reagent strip chemical
reactions are designed to perform at ionic
concentrations similar to urine
Interfering substances in the urine,
technical carelessness, and color
blindness also produces error
CONFIRMATORY TEST
Are defined as test using different reagent
or methodologies to detect the same
substances as detected by the reagent
strips with the same or greater sensitivity
or specificity
pH
Along with the lungs, kidneys are the
major regulators of the acid-base content
in the body
Normal first morning (slightly acidic): 5.06.0
Normal after meal (alkaline pH)
pH or random samples: 4.5-8.0
PROTEIN
the most indicative of renal disease
Proteinuria: is often associated with early
renal disease, making the urinary CHON
test important part of any physical
examination
Normal urine: less than 10 mg/dL or 100
mg per 24 hours is excreted

Albumin: major serum protein found in

normal urne
Tamm-Horsfall Protein (uromodulin):
produced by the renal tubular epithelial
cells; and proteins from prostatic, seminal
and vaginal secretions
CLINICAL SIGNIFICANCE
Clinical proteinuria: indicated at 30 mg/dL
or greater (300 mg/L)
Three major categories: prerenal, renal
and postrenal (based on the origin of the
urine)
Prerenal Proteinura
Caused by conditions affecting the plasma
prior to its reaching the kidney, and,
therefore is not indicative of actual renal
disease
Bence Jones Protein: primary example of
proteinuria due to increased protein levels;
seen in persons with multiple myeloma
Renal Proteinuria
Glomerular proteinuria: increased
amounts of serum protein and eventually
red and white blood cells pass through the
membrane and are excreted in the urine;
Increased pressure from the blood
entering the glomerulus may override the
selective filtration of the glomerulus,
causing increased albumin to enter the
filtrate
Microalbuminuria: presence of
microalbuminuria is also associated with
an increased risk of cardiovascular
disease
Orthostatic/Postural Proteinuria: frequent
in young adults; Increased pressure on
the renal vein when in the vertical position
is believed to account for this condition;
collect specimen upon rising in the
morning and after remaining in a vertical
position for several hours
Tubular Proteinuria: causes of tubular
dysfunction include exposure to toxic
substances and heavy metals, severe viral
infxns, and Fanconi syndrome
Postrenal Proteinuria

 Bacterial and fungal infxns and

inflammations produce exudates
containing protein from the interstitial fluid
REAGENT STRIP REACTIONS
Uses the principle of the protein error of
indicators to produce visible colorimetric
reaction
Indicator (yellow) + CHON (pH 3.0)
CHON + H+ ; indicator H+ (blue-green)
SULFOSALICYLIC ACID PPT TEST
SSA test is a cold precipitation test that
reacts equally in all forms of protein
TESTING FOR MICROALBUMINURIA
Random or first morning spx
Based on immunochemical assays for
albumin or albumin-specific reagent strips
Micral test and ImmunoDip: both are read
visually and first morning spx is required
Micral-Test reagent strips contain a goldlabeled antihuman albumin antibody
enzyme conjugate
ImmunoDip reagent strips uses
immunochromographic technique.
Albumin reagent strip: bis-3,4,5,6tetrabromosulphonphtalein; green to aqua
blue
Creatinine: Copper sulfate, 3,3,5,5tetramethylbenzidine, and diisopropyl
benzene dihydroyperoxide; orange though
green to blue; normally present in 10-300
mg/dL
GLUCOSE
Most frequently performed chemical
analysis on urine
Early diagnosis of diabetes mellitus
Renal threshold: 160 to 180 mg/dL
Specimen test: 2 hours after meal
Glycogenesis: glucose to glycogen
Glycogenolysis: gly to glu
Epinephrine: inhibitor of insulin secretion
REAGENT STRIP REACTION
Glucose oxidase provides a specific test
for glucose
Glucose to gluconic acid and peroxide

Peroxide and chromogen to oxidized

chromogen and water
COPPER REDUCTION TEST (Clinitest)
Earliest chem test performed on urine
Ability of glucose to reduce copper sulfate
to cuprous oxide in the presence of alkali
and heat
Negative blue green yellow
red/orange
KETONES
Increased fat metabolism include the
inability to metabolize CHO, as occurs in
DM, increased loss of CHO from vomiting
and associated with starvation
Ketonuria: shows a deficiency in insulin,
indicating the need to regulate dosage;
DM type 1
REAGENT STRIP REACTIONS
Uses Sodium Nitroprusside rxn
Acetone and B-hydroxybutyric acid are
produced from acetoacetic acid
Acetest: provides sodium nitroprusside,
glycine, disodium phosphate and lactose
in tablet form
BLOOD

Intact RBC = hematuria

RBC destruction = hemoglobinuria
Hematuria = cloudy red urine
Hemoglobinuria = clear red spx

Hematuria
Closely related to disorder of renal or
genitourinary origin in which bleeding is
the result of trauma or damage in the
organs
Severe back and abdominal pain are
suspected of having renal calculi
Strenuous exercise and menstruation =
nonpathologic
Hemoglobinuria
From lysis of RBC
Reabsorption of filtered hemoglobin also
results in the appearance of large yellow-

brown granules of denatured ferritin called

hemosiderin in the renal tubular epith.
Cells
Myoglobinuria
Heme-containing CHON found in muscle
tse
Associate with muscle destruction
REAGENT STRIP REACTIONS
Psuedoperoxidase activity of hemoglobin
False negative: menstruation, bacterial
enzymes, vitamin C
BILIRUBIN
Can provide early indication of liver
damage
Exhibited long before the patient exhibits
jaundice
Highly pigmented yellow compound,
degraded product of hemoglobin
REAGENT STRIP (DIAZO) RXNS
Uses diazo reaction
Bilirubin combines with 2,4-dicholoaniline
diazonium salt or 2,6-dichlorobenzenediazonium-tetrafluoroborate in an acid
medium to produce azodye
Tan or pink to violet (color range)
False negative: spx are not fresh
Ictotest: confirmatory test for bilirubin
UROBILINOGEN
Intestinal bacteria convert bilirubin to a
combination of urobilinogen and
stercobilinogen.
Responsible for the brown color of feces
CLINICAL SIGNIFICANCE

Increased urine urobilinogen is seen in

liver dse and hemolytic disorders

NITRITE
Rapid screening test for the presence of
UTI
Determining initial bladder infection
REAGENT STRIP RXN
Detected by Greiss reaction; nitrite at
acidic pH reacts with aromatic amine
INTERFERENCES
Bacteria do not possess te ability to
reduce nitrate to nitrite
Bacteria capable of reducing nitrate must
remain intact with the urinary nitrate
Realibility of results depends on the
amounts of nitrate in the urine
Further reduction of nitrite to nitrogen may
occur
False negative results: bacterial
metabolism, ascorbic acid, decreased
sensitivity in specimens
LEUKOCYTE ESTERASE
Detection of urinary leukocyte
Normal values: 0-2 to 0-5/hpf
Detects WBC except lymphocyte
REAGENT STRIP REACTION
Uses the action of LE to catalyze the
hydrolysis of an acid ester embedded on
the reagent pad to produce an aromatic
compound and acid
REACTION INTERFERENCE
Presence of strong oxidizing agents
High conc of CHON, glucose, oxalic acid
and ascorbic acid
SPECIFIC GRAVITY
Measures only ionic solutes

REAGENTS STRIP RXN & INTERFERENCES

Multistix uses Ehrlichs aldehydes rxn;

color ranges from light to dark pink
Chemstrip incorporates an azo-coupling
rxn using 4-methoxybenzene diazoniumtetrafluoroborate

MICROSCOPIC EXAMINATION
- Least standardized
- Results depend on the skill of microscopist
PREPARATION OF URINE SEDIMENT

Urine volume: 12 mL
Speed of centrifugation: 400 RCF
RCF = (1.118)(10^-5)(radius in cm)(RPM2)
Overcentrigugation: cellular disintegration
Undercentrifugation: less sediments recovered
Time of centrifugation: 5 mins
Volume of sediment: 20uL
METHOD: STANDARDIZATION/ COMMERCIAL
SYSTEM using conical centrifuge automatic 1
mL when decanted
REPORTING OF RESULTS

STAINING TECHNIQUES
***check book
RBC
-

RBC, WBC, Casts: average no.

EC, crystals: qualitative description

Rare

Hard to find

Few

Moderate

7-8 um in diameter
Smooth, non-nucleated, biconcave disc
Normal: 0-3/hpf
Higher: pathogenic
Hypersterunic urine: crenated (high SG)
Hyposterunic urine: ghost cells membrane
remains intact; low refractive index
Dysmorphic RBC: glomerular bleeding
acanthocytes
Most often confused with yeast cells (budding),
Oil droplets (too big, vary sizes, high refractive
index), WBC (granules)
If still not distinguished from others ADD 2%
ACETIC ACID RBC would lyse, WBC
become prominent, yeast still intact
Correlations:
Physical: Color:reddish, pinkish, brownish
Chemical: blood rxn (ascorbic acid interference),
renal dse, low urinary tract (acute and chronic
infxn), extra renal dse (acute appendicitis), toxic
rxn (drugs), physiologic causes (exercise)
WBC

- Normal: 0-5/hpf
- Too much water: BROWNIAN MOVEMENT in
hypotonic urine: Glitter cells
- Renal Tubular EC are bigger & single
eccentrically central nucleus
- Correlation: LE & Nitrite Rxn are (+)
- Reasons: 1. Bacterial infxn or inflammation of
genito-urinary tract 2. Bladder tumor 3. Fevers
and strenuous exercise
- Neutrophil predominant
EPITHELIAL CELLS

Many

- Normal but little

SQUAMOUS EC

TNTC

Not significant, but if high = contamination

Abundant and prominent nucleus
Common in female urine
Gardnerella vaginalis, affect cytoplasm of EC
Clue cell dirty cell
TRANSITIONAL

- Caudate or polyhedral

- Bladder, ureter, urethra

- Viral and malignant infxn if high

- Confused with granular cast (differentiate by gram

staining)

RENAL TUBULAR CELLS

-

Originate from renal tubules

Eccentrically located nucleus
Normal: 2RTE/hpf
> 2 RTE/hpf: necrosis and toxicity
Oval fat bodies: RTE with lipid inclusion:
Nephrotic syndrome
- Bubble cell: RTE with non lipid vacuole
CASTS
-

Tubular lumen, where it is formed

Uromodulin, core matrix
Unique to kidneys: DCT & collecting tubules
Process specimen right away
Cylindroids
Parallel sides, sometimes tapered end
How is cast formed: Acid pH, increased solute
conc, urinestasis, increased plasma protein
(whatever solute in excess that would be type of
cast)
TYPES OF CAST:
HYALINE CAST

Very low refractive index

0-2/lpf
Strenuous exercise
Acute glomerulonephritis, chronic renal dse, CHD

GRANULAR CAST
- Made up of uromodulin
- Stress, dehydration, strenuous exercise

CRYSTALS
- Formation: pH of urine, conc of solute, flow of
urine in tubules (disruption)
- Enhanced in refrigeration
ACIDIC URINE
- Amorphous Urate: long standing, brick dust
(orange pink precipitate)
- Uric acid
- Sodium urate
- Calcium oxalate dehydrate or monohydrate
ALKALINE URINE
- Amorphous Phosphate: long standing, white to
beige precipitate
- Triple Phosphate (Mg, Ammonium Phosphate;
coffine lid)
- Calcium Phosphate
- Mg Phosphate
- Ammonium Biurate: thorny apple
- Ca Carbonate: dumbbell shape

WAXY CAST
-

Broken edges, blot ends, serated ends

Highly refractive
Ground glass appearance
Indicates poor prognosis
From degeneration of other casts
Aka renal failure cast
Only form in extreme urinestasis
Cellular granular waxy broad cast

ABNORMAL
-

RBC CAST
- Yellow/orange pigment
- Freely floating RBC around it
- Glomerulonephritis, acute tubular necrosis
WBC CAST
- Help us differentiate ordinary UTI or lower UTI
with infxn in kidney called Pyelonephritis
BACTERIAL CAST

Signify pathologic condition

Hexagonal plate: CYSTEINE
Butch plate: cholesterol
Radiographic dye
Tyrosine
Leucine
Bilirubin (liver disorder)
Sulfonamide (medication)
OTHER ELEMENTS
MICROORGANISMS

Bacteria
Yeast candida albicans diabetic patients
Trichomonas vaginalis parasite, itching vagina
Clue cells and Gardnerella vaginalis
MISCELLANEOUS

- Mucus threads low refractive index

- Sperm dont report

- Contaminants such as fiber, starch, oil droplets,
air bubbles, pollen grains dont report