MICROSOPIC EXAMINATION OF THE URINE

"MACROSCOPIC SCREENING AND MICROSCOPIC SCREENING"

Test Significance
• COLOR • Blood
• CLARITY • Hematuria vs. Hemoglobinuria
• BLOOD • RBC,RBC casts
• PROTEIN • Casts,cells
• NITRITE • Bacteria,WBCs
• LE • WBCs,WBC casts, bacteria
• GLUCOSE • Yeast cells

HOW SPECIMEN IS PREPARED? SPECIMEN VOLUME AND

 Fresh urine/midstream urine is a CENTRIFUGATION
must. At 37C • 10-15mL- volume needed for
 Don’t refrigerate sample (may centrifugation in conical tube
cause precipitation) • 12 mL-for reagent dip stick test, done
 Dilute specimen any cause false first
negative results. • 5 minutes in 400 rcf - recommended
in cetrifugation . Calibration must be
done first.

HOW TO PREPARE URINE VOLUME OF SEDIMENT

SEDIMENT? EXAMINED
▫ Volume needed is 0.5-1 mL • If using glass-slide method
▫ To maintain concentration, 20 uL (0.02 mL) covered by 22x22
centrifuged urine must be mm glass cover slip , allowing
aspirated off than poured specimen to flow.
off/decant. • Read at microscope a
▫ Sediment must resuspended by minimum of 10 fields under
gentle agitation-by tapping the tip both hpf and lpf using Bright -
of the tube with a finger . field microscope.
Cover slip
▫ Avoid vigorous agitation, it
disrupts cellular elements.

HOW TO REPORT MICROSCOPIC

EXAMINATION?
> Lpf-CASTS,EPITHELIAL
CELLS,URATES AND PHOPHATES,
crystals
> Hpf-RBC, WBC, parasites,bacteria,
mucus threads,yeast cells
> Minimum of 10 fields both

STAINING EXAMINATION TECHNIQUES

→ Staining increases visibility of the cells and other sediments in urine. Under bright field microscopy by changing
refractive index.
→ Most frequently used is the Sternheimer-Malbin stain.
STERNHEIMER-MALBIN FOR CONTRASTING COLORS OF NUCLEUS AND CYTOPLASM WBC,EPITHELIAL CELLS,CASTS
TOLUIDINE BLUE ENHACES NUCLEAR DETAIL DIFFERENTIATES WBC from RTE cells
2% ACETIC ACID LYSES RBC AND ENHANCE NUCLEUS OF WBC DISTINGUISHES RBC FROM WBC,YEAST, OIL
DROPLETS,AND CRYSTALS
OIL RED O AND SUDAN III STAINS TRIGLYCERIDE BUT NOT CHOLESTEROL LIPID CONTAINING CELLS AND CASTS
GRAM STAIN DIFFERENTIATES GRAM POSITIVE AND NEGATIVE BACTERIA IDENTIFIES BACTERIAL CASTS
HANSEL STAIN METHYLENE BLUE AND EOSIN Y GRANULES FOR EOSINOPHILS
PRUSSIAN BLUE STAINS IRON-CONTAINING STRUCTURES HEMOSIDERIN

URINE MICROSCOPY PART 1 Page 1

 URINALYSIS MICROSCOPIC TECHNIQUES
BRIGHT-FIELD MICROSCOPY For routine urinalysis

PHASE CONTRAST MICROSCOPY Enhances low refractive index -Dysmorphic RBC causes: glomerular bleeding
sediments (Hyaline cast,mucous -WBC causes: Pyuria, Inflammation (suffix "is")
-Clue cell causes: Gardnerella vaginalis
threads,trichomonas) -High Urothelium another term: synctia
-RTE causes: tissue destruction,drugs,hepaB, salicylate
POLARIZING MICROSCOPY For identification of lipid Poisoning,,acute transplant rejection,myoglobin
containing sediments toxicity
-Oval fat bodies-tubular necrosis,DM,trauma
-Bacteria causes: UTI either staph or E.coli
DARK-FIELD MICROSCOPY Identification of treponema -Yeast causes:Candida , diabetes, vaginal monaliasis
pallidium -Parasites: Trichomonas,Schistosoma haematobium
-Mucus is a protein produced by glands and
FLOURESCENCE MICROSCOPY Identification of labeled antigens Epithelial cells in lower genitourinary tract and RTE
and antibodies cells

INTERFERENCE MICROSCOPY Produces 3D microscopy

"COMMON URINE SEDIMENTS SEEN IN MICROSCOPE "
NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS
RBC Anucleated Yeast cells Average # per 10 Color
Crenated in hypertonic urine Oil droplets HPF Blood +
Ghost cells in hypotonic urine Air bubbles
Dysmorphic in shape during
glomerular damage

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS

WBC Larger than RBCs RTE cells Average # per 10 LE +
Granulated, multilobed HPF Nitrite + WBC
neutrophils (most common) Spgv (high)
Mononuclear cells with PH (high)
abundant cytoplasm

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATION

S
SQUAMOUS Largest, with abundant Casts Rare,few, Clarity
EPITHELIAL irregular cytoplasm, moderate or many
CELLS prominent nuclei per LPF
TRANSITION AL Spherical,polyhedral,caudat Spherical resembles Rare,few, Clarity
EPITHELIAL e, centrally located nucleus RTE cells moderate or many Blood +
CELLS per LPF
RTE cells Rectangular, columnar (by Spherical Average # per 10 Color
PCT) round and transitional HPF Clarity
oval,cuboidal (by DCT) with epithelial cells LE +
eccentric nucleus . Granular cast Nitrite +
Protein +
Bilirubin +
Blood +

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS

Oval Fat High refractile RTE cells Fat stains Average # per 10 Color
Bodies HPF Blood +
Protein
Fatty casts

Oval fat bodies

URINE MICROSCOPY PART 1 Page 2

 "MISCELLANEOUS SEDIMENTS SEEN IN MICROSCOPE "

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS

Bacteria Small,spherical,rod shaped Amorphous Rare,few,moderat pH (high)
phosphates and e,or many with Nitrite +
urates WBC per HPF LE + bacteria
WBC

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS

Yeast Small.oval,refractile with RBC Rare,few,moderat Glucose +
buds or mycelia e,or many with LE +
WBC per HPF WBC

Yeast (Candida albicans )

Yeast cells

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS

Trichomo Pear WBC,RTE Rare,few,moderat LE + OTHER INFO:
nas shaped,motile,flagellated e,or many with WBC -Another parasite named
WBC per HPF Schistosoma
haematobium can also be
seen in urine

NAME APPEARANCE SOURCES OF REPORTING CORRELATIONS

ERROR
Sper Tapered oval head with None Based on lab Protein +
matozoa long , thin tail protocol

NAME APPEARANCE SOURCES OF ERROR REPORTING CORRELATIONS

Mucus Single, clumped threads with Hyaline casts Rare,few,moderat None
a low refractive index e,or many with
WBC per HPF

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