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

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


CLINICALLY SIGNIFICANT!
 Not normally present in urine especially if specimen  Type: RBC
is taken under normal condition.  Description: RBCs in cast matrix. Yellowish to
 Most indicative of urinary tract infection orange color or orange-red color
 Most common cause of UTI are the Enterobacters  Significance: Acute glomerulonephritis. Strenous
exercise
Yeast  Comments: Pinpoints source of bleeding in kidney.
 Small, refractile oval structures, usually mistakes as Most fragile of casts. Often in fragments.
RBC
 Candida albicans is the most common cause of yeast  Type: Blood
UTI.  Description: Contain hemoglobin. Yellowish to
CLINICAL SIGNIFICANCE orange color.
 Diabetes  Significance: Same as RBC cast
 Immunocompromised patients  Comments: From disintegration of RBC casts
 Vaginal Moniliasis
 Type: WBC
Parasites  Description: Leukocytes incorporated into cast
Trichomonas vaginalis matrix. Irregular in shape.
 The most frequent parasite encountered in urine  Significance: Pyelonephritis
samples  Comments: Pinpoints kidneys as the site of infection
 Resmbles WBC, transitional cells or RTE; hard to
identify when not moving.  Type: Epithelial cell
 Can be seen in wet mount preparation  Description: Renal tubular epithelial cells
incorporated into cast matrix
Schistosoma haematobium  Significance: Renal tubular damage
 Parasite of the urinary bladder that can be excreted  Comments: Transitional and squamous epithelial
in urine. cell casts do not exist. These cells are found distal to
renal tubules and collecting ducts where casts are
Enterobius vermicularis formed.
 Most common ova contaminant in urine
 Type: Waxy
Spermatozoa  Description: Homogenous, opaque, notched edges,
 Oval, slightly tapered heads with long, flagella- like broken ends
tails.  Significance: Urinary stasis
 Occasionally found in both male and female following  Comments: From degeneration of cellular and
sexual intercourse granular casts. Unfavorable sign.
 Not usually reported in the laboratory due to possible
legal consequences  Type: Fatty
 Description: Casts containing lipid droplets
Mucus  Significance: Nephrotic syndrome
 Thread-like structure that is made of protein  Comments: Maltese crosses with polarized light.
materials Stain with Sudan and oil red O.

TAMM-HORSFALL PROTEIN  Type: Broad


 Produced at the distal convoluted tubule by RTE  Description: Wide. May be cellular, granular, or
excretion waxy.
 Major component of mucus and casts.  Significance: Advanced renal disease
 Comments: Formed in dilated distal tubules and
Casts collecting ducts. “Renal failure casts.”
 The only element found in the urinary sediments
 that are unique to the kidney Crystals in Acidic/Neutral Urine
 Made up of TAMM-HORSFALL PROTEIN
 Cannot be detected by reagent strips, therefore,  Crystal: Amorphous urates
increased or positive CHON strip is frequently  Description: Irregular granules
associated with renal disease.  Significance: None
 Comments: From pink precipitate in bottom of tube.
CYLINDURIA May obscure significant sediment. Dissolved by
 Presence of casts in urine warming to 60ºC.

 Type: Hyaline  Crystal: Uric acid


 Description: Homogenous with parallel sides and  Description: Very pleomorphic. Four-sided, six-
rounded ends sided, star-shaped, rosettes, spears, plates.
 Significance: 0-2/low-power field (LPF) are normal. Colorless, red brown, or yellow.
Increased with stress. Fever, trauma, exercise, renal  Significance: Usually normal
disease  Comments: Birefringent. Polarized light.
 Comments: Most common type. Least significant.
Contain Tamm-Horsfall protein only . May be  Crystal: Calcium oxalate
overlooked if light is too bright  Description: Octahedral (eight-sided) envelope
form is most common. Also dumbbell and ovoid
 Type: Granular forms.
 Description: Same as hyaline, but contains granule  Significance: Normal
 Significance: 0-1/LPF is normal. Increased with  Comments: Occasionally found in slightly alkaline
stress, exercise, glomerulonephritis, Pyelonephritis. urine. Monohydrate form may be mistaken for RBCs.
 Comments: May originate from disintegration of Most common constituent of renal calculi.
cellular casts.
 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