Microscopic Analysis of 


Enhances nuclear detail of cells
Differentiates WBCs from RTE cells

Urine  Useful for cell identification of other body fluids

3. 2% Acetic Acid
Microscopic Urine Analysis  Lyses RBCs
 Third part of routine urinalysis  Enhances nuclear detail WBCs.
 Used to detect/ identify/ quantify insoluble materials
present in urine (sediments, crystals, cells) 4. Hansel Stain
 Most time consuming part of routine urinalysis  Composed of methylene blue and eosin Y
 LEAST STANDARDIZED among all analyses  Preferred stain for urinary eosinophls
conducted in urine sample.  Dried smear of centrifuged urine is used
 Also known as chemical seiving.
5. Wright’s Stain
Urine Sediment Preparation and Examination  Primarily used for peripheral blood smear but can
 Standardization started in 1926 by Thomas Addis also be use as a substitute for urinary staining
(Addis’ Count) (particularly eosinophil)
- uses hemocytometer  Not as specific as Hansel stain for eosinophil
- 12 hour urine sample
- Used to monitor the course of diagnosed cases of 6. Prussian Blue Stain
RENAL DISEASE.  Stains hemosiderin granules as seen in
 Commercialized methods have been developed hemoglobinuria
throughout the years.  Hemosiderin granules stain BLUE
e.g.
KOVA CenSlide Lipid Stains
Urisystem R/S Workstation 1. Oil red O, Sudan III & Sudan IV
Count-10 Quick-Prep  For the identification of lipid containing cells and free
fat bodies.
CenSlide  Triglycerides and neutral fats stains well with a
 Doesn’t require manual charging of urine sample on ORANGE-RED color
slides
 Closed-system procedure to eliminate exposure Bacterial Stain
 Especially designed tube to facilitate urine 1. Gram’s Stain
microscopy  Differentiates Gram positive from Gram negative
bacterial infection
R/S Workstation  For bacterial cast identification
 Glass flow cell through which urine sediment is
pumped, examined and discarded from the system. 2. Ziehl Neelsen Stain
 Differentiates acid fast organisms from non acid fast
Specimen Preparation organisms.
 Freshly collected then thoroughly mixed, midstream  For urinary tubercular infection
clean-catch sample
 10-15 mL (standard amount of urine ) Cytodiagnostic Testing
 Centriguation time: 5 minutes at 400 RCF 1. Papanicolaou’s Stain
 Decantation (rapid pouring of centrifuged urine  Used for preparing fixed slides for urine sediment
sample) identification and cytology.
 Charging the sediment on the slide (20uL/ 0.02 mL)  For the detection of malignancies in lower urinary
 Examination under the microscope tract.
 Reporting the microscopic exam semiquantitatively  Detects and monitors renal diseases
(rare, few, moderate, many)  Provides more definitive information about:
transplant rejection, inflammatory conditions,
Checking the Correlation… pathologic casts
Correlation between physical and chemical exam plus the  First morning urine sample or suprapubic aspirate.
microscopic exam must be observed to ensure accuracy and
reliability of reports. MICROSCOPE
Bright-field Microscopy
Sediment Examination Techniques  Most common type of microscopy done in the lab
STAINS  Object appears dark against light background
 Increases the overall visibility of sediments being  When using this method, decreased light level must
analysed. be employed. (Adjust the rheostat level)
 Imparts identifying characteristics to cellular
structures: nuclei, cytoplasm, and inclusions. Phase-Contrast Microscopy

Sediment/ Cellular Stain

1. Sternheimer-Malbin Stain
 Supravital stain (stains living cells)
 Made up of crystal violet and safranin O
 Absorbed well by WBC, EC, Cast
 Provides clearer delineation of cells
 Provides contrasting color of the nucleus and
cytoplasm

2. 0.5% Solution of Toluidine Blue

 Metachromatic stain
Polarizing Microscopy
 Confirms the identification of fat droplets, oval fat
bodies and fatty casts that produce MALTESE CROSS
APPEARANCE
Interference Contrast Microscopy
 Provides a three dimensional image by showing very
fine structural detail
 ADVANTAGE: object appears dark against light
background without any halo associated with phase
contrast microscopy
Dark-Field Microscopy
 Enhances visualization of specimens that cannot be
seen easily with bright field microscopy
 For identification of spirochetes (Treponema,
Borrelia, etc)
Fluorescence Microscopy
 Used to detect bacteria and viruses within cells and
tissues through immunofluorescence.
 FLUORESCENCE: property by which an atom
absorbs light of a particular wavelength and emit
light of a longer wavelength, fluorescence lifetime
Urine Sediments
Red Blood Cells
 Smooth, non-nucleated, biconcave discs
 7um in diamter
 MOST DIFFICULT to recognize by students because
of variation in sizes (anisocytes), characteristic
structures and close resemblance to YEAST,
BUBBLES, OIL DROPLETS and other microscopic
sediments.
Mononuclear cells
 Not identified in wet mount preparation of urine
 LYMPHOCYTES
- resembles RBCs
- increased in early stages of renal rejection
 MONOCYTES, MACROPHAGE, HISTIOCYTES
- large cells with vacuoles
The primary concern of mononuclear cells is to
differentiate them from RTE
CLINICAL SIGNIFICANCE
 Pyuria (increased WBC in urine)
 Pyelonephritis (ascending UTI; involve the kidneys)
 Bacteuria (increased bacterial multiplication)
 Cystitis (inflammation of the bladder)
Epithelial Cells
 Normally seen in urine except in large amounts or
abnormal forms
3 Types of EC:
1. Squamous EC
2. Transitional EC (urothelial cells)
3. Renal Tubular Cells
Squamous Epithelial Cells
 Largest cells found in urine sediment
 Originates from the linings of vagina and female
urethra, and the lower portion of the male urethra.
CLUE CELLS
 infected epithelial cells
 Seen in bacterial vaginosis
 Gardnerella vaginalis

HYPERSTHENURIC SAMPLE: Transitional Epithelial Cells

 RBCs shrink= CRENATED= loss of water  Originates from the lining of renal pelvis, calyces and
HYPOSTHENURIC SAMPLE: bladder
 RBCs swell, lyse, relase Hgb= GHOST CELLS  Smaller than epithelial cells
 Spherical, polyhedral or caudate in shape due to the
DYSMORPHIC RBC ability to absorb water.
 Aids in determining the site of renal bleeding  Pathologically seen in Malignancy and Viral Infection
 Associated with glomerular bleeding
 ACANTHOCYTES SYNCTIA
 increased numbers of transitional cells seen in
CLINICAL SIGNIFICANCE clumps.
 Glomerular bleeding  Seen in catheterization collection
 Lower urinary tract bleeding  Considered as NORMAL
 Trauma
 Acute infection/ inflammation Renal Tubular Epithelial cells
 Coagulation disorder  Slightly larger than WBC
 May be flat, cuboidal, columnar
MANNER OF REPORTING:  Must be identified under HPO
/ hpf  THE MOST CLINICALLY SIGNIFICANT URINARY
EPITHELIAL CELL (e.g. oval fat bodies)
White Blood Cells
 Larger than RBCs CLINICAL SIGNIFICANCE
 12 mm in diameter  Malignancy
 Stains well with Sternheimer-Malbin Stain  Exposure to heavy metals
 NEUTROPHIL- predominating urinary WBC  Tubular damage
At HYPOTONIC ENVIRONMENT = SWELL  Pyelonephritis
BROWNIAN MOVEMENT = GLITTER CELLS  Kidney transplant rejection

Eosinophil Oval Fat Bodies

 Stains well with Hansel Stain
 Seen in:
- Drug- induced interstitial nephritis (allergy)
- UTI (parasitic in origin)
- Renal transplant rejection (anaphylactic
reaction)
 RTE cells that absorb lipid
 “lipid containing RTEC”
 Lipiduria
- presence of lipid/ fat in urine
- Most frequently associated with nephrotic
syndrome

NOT NORMALLY SEEN IN URINE= (at least 1%) Bacteria

CLINICALLY SIGNIFICANT!
  Not normally present in urine especially if specimen
is taken under normal condition.
 Most indicative of urinary tract infection
 Most common cause of UTI are the Enterobacters
Yeast
 Small, refractile oval structures, usually mistakes as
RBC
 Candida albicans is the most common cause of yeast
UTI.
CLINICAL SIGNIFICANCE
 Diabetes
 Immunocompromised patients
 Vaginal Moniliasis
Parasites
Trichomonas vaginalis
 The most frequent parasite encountered in urine
samples
 Resmbles WBC, transitional cells or RTE; hard to
identify when not moving.
 Can be seen in wet mount preparation
Schistosoma haematobium
 Parasite of the urinary bladder that can be excreted
in urine.
Enterobius vermicularis
 Most common ova contaminant in urine
Spermatozoa
 Oval, slightly tapered heads with long, flagella- like
tails.
 Occasionally found in both male and female following
sexual intercourse
 Not usually reported in the laboratory due to possible
legal consequences
Mucus
 Thread-like structure that is made of protein
materials
TAMM-HORSFALL PROTEIN
 Produced at the distal convoluted tubule by RTE
excretion
 Major component of mucus and casts.
Casts
 The only element found in the urinary sediments
 that are unique to the kidney
 Made up of TAMM-HORSFALL PROTEIN
 Cannot be detected by reagent strips, therefore,
increased or positive CHON strip is frequently
associated with renal disease.
CYLINDURIA
 Presence of casts in urine
 Type: Hyaline
 Description: Homogenous with parallel sides and
rounded ends
 Significance: 0-2/low-power field (LPF) are normal.
Increased with stress. Fever, trauma, exercise, renal
disease
 Comments: Most common type. Least significant.
Contain Tamm-Horsfall protein only . May be
overlooked if light is too bright
 Type: Granular
 Description: Same as hyaline, but contains granule
 Significance: 0-1/LPF is normal. Increased with
stress, exercise, glomerulonephritis, Pyelonephritis.
 Comments: May originate from disintegration of
cellular casts.
 Type: RBC
 Description: RBCs in cast matrix. Yellowish to
orange color or orange-red color
 Significance: Acute glomerulonephritis. Strenous
exercise
 Comments: Pinpoints source of bleeding in kidney.
Most fragile of casts. Often in fragments.
 Type: Blood
 Description: Contain hemoglobin. Yellowish to
orange color.
 Significance: Same as RBC cast
 Comments: From disintegration of RBC casts
 Type: WBC
 Description: Leukocytes incorporated into cast
matrix. Irregular in shape.
 Significance: Pyelonephritis
 Comments: Pinpoints kidneys as the site of infection
 Type: Epithelial cell
 Description: Renal tubular epithelial cells
incorporated into cast matrix
 Significance: Renal tubular damage
 Comments: Transitional and squamous epithelial
cell casts do not exist. These cells are found distal to
renal tubules and collecting ducts where casts are
formed.
 Type: Waxy
 Description: Homogenous, opaque, notched edges,
broken ends
 Significance: Urinary stasis
 Comments: From degeneration of cellular and
granular casts. Unfavorable sign.
 Type: Fatty
 Description: Casts containing lipid droplets
 Significance: Nephrotic syndrome
 Comments: Maltese crosses with polarized light.
Stain with Sudan and oil red O.
 Type: Broad
 Description: Wide. May be cellular, granular, or
waxy.
 Significance: Advanced renal disease
 Comments: Formed in dilated distal tubules and
collecting ducts. “Renal failure casts.”
Crystals in Acidic/Neutral Urine
 Crystal: Amorphous urates
 Description: Irregular granules
 Significance: None
 Comments: From pink precipitate in bottom of tube.
May obscure significant sediment. Dissolved by
warming to 60ºC.
 Crystal: Uric acid
 Description: Very pleomorphic. Four-sided, six-
sided, star-shaped, rosettes, spears, plates.
Colorless, red brown, or yellow.
 Significance: Usually normal
 Comments: Birefringent. Polarized light.
 Crystal: Calcium oxalate
 Description: Octahedral (eight-sided) envelope
form is most common. Also dumbbell and ovoid
forms.
 Significance: Normal
 Comments: Occasionally found in slightly alkaline
urine. Monohydrate form may be mistaken for RBCs.
Most common constituent of renal calculi.
  Crystal: Leucine
 Description: Yellow, oily-looking spheres with radial
and concentric striations
 Significance: Severe liver disease
 Comments: Often accompanied by tyrosine

 Crystal: Tyrosine
 Description: Fine yellow needles in sheaves or
rosettes
 Significance: Severe liver disease
 Comments: Often accompanied by leucine

 Crystal: Cystine
 Description: Hexagonal (six-sided)
 Significance: Cystinuria
 Comments: Must be differentiated from uric acid.
Does not polarize light.

 Crystal: Cholesterol
 Description: Flate plate with notched out corner.
“Star-step.”
 Significance: Nephrotic syndrome
 Comments: Birefringent

 Crystal: Bilirubin
 Description: Yellowish-brown needles, plates, and
granules
 Significance: Liver disease
 Comments: Reagent strip or Ictotest should be
positive for bilirubin.

Crystals Found in Alkaline Urine

 Crystal: Amorphous phosphates

 Description: Irregular granules
 Significance: None
 Comments: Form white precipitate in bottom of
tube. Dissolve with 2 acetic acid.

 Crystal: Triple phosphate

 Description: “Coffin-lid”crystal
 Significance: None

 Crystal: Ammonium biurate

 Description: Yellow-brown “thorn apples” and
spheres
 Significance: None
 Comments: Seen in old specimens

 Crystal: Calcium phosphate

 Description: Needles, rosettes, “pointing finger”
 Significance: None
 Comments: Only needle form seen in alkaline urine

 Crystal: Calcium carbonate

 Description: Colorless dumbbells
 Significance: None