AUBF311: ANALYSIS OF URINE & BODY FLUIDS

Lecture Module First Semester

Week 05 S.Y. 2023-2024

MICROSCOPIC EXAMINATION OF URINE

TOPIC OUTLINE URINE SEDIMENTS
1 Macroscopic Screening
2 Urine Sediments  variety of formed elements (RBCs, WBCs, ECs,
Bacteria, Yeast, Parasites, Spermatozoa, Casts,
Crystals, Artifacts)
MACROSCOPIC SCREENING

SPECIMEN VOLUME COMMONLY LISTED VALUES

10-15 mL standard RBCs 0-2 or 3 / HPF
12 mL average WBCs 0-5 or 8 / HPF
Hyaline Casts 0-2 / LPF
CENTRIFUGATION
 400 RCF for 5 minutes RED BLOOD CELLS
 smooth, non-nucleated, biconcave disks
SEDIMENTATION PREPARATION  manner of reporting: average number per 10 hpf
 decantation
0.5-1.0 mL standard
. In Concentrated
RBCs shrink due to loss of
(Hypersthenuric)
VOLUME OF SEDIMENT EXAMINED water, & appear crenated
Urine
 recommended: 20 µL (0.02 mL) covered by 22 x In Dilute RBCs absorb water, swell,& lyse
22 mm glass cover slip (Hyposthenuric) releasing Hgb & leaving only cell
Urine membrane becoming ghost cells
REPORTING MICROSCOPIC EXAMINATION
Casts average number per LPF (10 fields) SOURCES OF ERRORS
RBC RBCs lack of characteristic structures, variations
WBC
average number per HPF (10 fields) 
in size & close resemblance to other constituents
Epithelial RBCs are confused with yeast cells, oil droplets,
Cells rare, few, moderate, many 
& air bubbles
Crystals
Other
1+, 2+, 3+, 4+
Elements

ADDIS COUNT
 standardized quantitation of formed elements in
urine microscopic analysis developed by Addis in
1926 using hemocytometer
12-hour Specimen specimen
NORMAL VALUES
RBCs 0-500,000
WBCs DYSMORPHIC CELLS
0-1,800,000
Epithelial Cells  RBCs that vary in size, have cellular protrusions,
Hyaline Casts 0-5,000 or are fragmented
 associated primarily with glomerular bleeding
.

WRIGHT’S STAIN
NOTE: Formalin is used as preservative but not with
 show cell to be hypochromic & better delineate
glucose determination
.
presence of cellular blebs & protrusions

NOTE: 2% Acetic Acid lyses RBCs

College Of Medical Laboratory Science │Ranz Barron Bamba

 AUBF311: ANALYSIS OF URINE & BODY FLUIDS
Lecture Module First Semester
Week 05 S.Y. 2023-2024

EOSINOPHILS
 associated with drug-induced interstitial nephritis
(acute interstitial nephritis)
 small numbers are seen with UTI & renal
transplant rejection
 preferred stain: Hansel stain
>1% considered significant
.
<1% normal values

Macroscopic
cloudy with a red to brown color
Hematuria
Microscopic >100 / HPF
Hematuria early diagnosis of glomerular disorder

WHITE BLOOD CELLS

 larger than RBCs
 associated with pyelonephritis
 manner of reporting: average number per 10 hpf
MONONUCLEAR CELLS
SOURCES OF ERRORS smallest WBC & resembles RBCs
 renal tubular epithelial (RTE) cells Lymphocytes increased in early stages of renal
transplant rejection
NOTE: prolonged standing of alkaline urine lyse WBC Monocytes
large cells & appear vacuolated or
Macrophages
contain inclusions
Histiocytes

PYURIA
 increase in urinary WBCs
 indicates presence of infection or inflammation in
genitourinary system
 frequently caused by bacterial infections
(pyelonephritis, cystitis, prostatitis & urethritis)
NEUTROPHILS
 predominant WBC EPITHELIAL CELLS
 contain granules & multilobed nuclei
 lyse rapidly in dilute alkali urine  not unusual in urine
 derived from linings of genitourinary system
GLITTER CELLS
 neutrophils exposed to hypotonic urine absorb
water & swell, Brownian movement of granules
produce sparkling appearance
STERNHEIMER-MALBIN STAIN
.
 stains light blue

SQUAMOUS EPITHELIAL CELLS

 largest cells found in urine sediment
 first structure observed under low-power
magnification
 good reference for focusing microscope
 manner of reporting: RFMoMa

College Of Medical Laboratory Science │Ranz Barron Bamba

 AUBF311: ANALYSIS OF URINE & BODY FLUIDS
Lecture Module First Semester
Week 05 S.Y. 2023-2024

ORIGIN
 linings of vagina & female urethra
.
 lower portion of male urethra

CLUE CELLS
 variation of squamous epithelial cells
 indicative viral infection by Gardnerella vaginalis
 bacteria cover most of cell surface (70%) &
extend beyond edges of cells

CLINICAL SIGNIFICANCE
TRANSITIONAL EPITHELIAL (UROTHELIAL) CELL  most clinically significant epithelial cells
 smaller than squamous cells  indicative of necrosis of renal tubules
 spherical, polyhedral, & caudate
 present in small numbers in normal urine FILTRATES ABSORBED BY RTE CELLS
 increased in catheterization
results of liver damage
 manner of reporting: RFMoMa Bilirubin
appear a deep yellow color
ORIGIN
converted to hemosiderin
 lining of renal pelvis, calyces, ureter, & bladder appear yellow-brown hemosiderin
 upper portion of male urethra Hemoglobin
. granules
may be seen free-floating
RENAL TUBULAR EPITHELIAL CELLS
Proximal larger than other RTE PRUSSIAN BLUE STAIN
Convoluted rectangular & referred as columnar  confirmation of the presence of hemosiderin
Tubule (PCT) resembles casts  iron-containing hemosiderin granules stain blue
smaller than those from PCT
Distal
round or oval
Convoluted
Tubule (DCT) mistaken for WBCs & spherical
transitional epithelial cells
cuboidal & never round
eccentrically placed nucleus
Collecting
mistaken for granular casts
Ducts
manner of reporting: average
number per 10 HPF

College Of Medical Laboratory Science │Ranz Barron Bamba

 AUBF311: ANALYSIS OF URINE & BODY FLUIDS
Lecture Module First Semester
Week 05 S.Y. 2023-2024

BACTERIA Schistosoma haematobium

 bladder parasite
 multiply rapidly in room temperature for
 associated with bladder cancer
extended periods
 produce positive nitrite test & pH >8
(unacceptable specimen)
 present in a form of cocci (spherical) or bacilli
(rods)
 significant for UTI should be accompanied by
WBCs
 manner of reporting: RFMoMa

Enterobius vermicularis
 fecal contamination

YEAST
SPERMATOZOA
 small, refractile oval, may be branched
 have budding cells  oval, slightly tapered heads, & long, flagella-like
 true yeast infection must be accompanied by tails
presence of WBCs  found in urine of both men & women after sexual
intercourse, masturbation, or nocturnal emission
Candida albicans  significant in cases of male infertility or
 seen in urine of diabetic or immunocompromised retrograde ejaculation (sperm is expelled into
patients & women with vaginalis moniliasis bladder instead of urethra)
Germ Tube (+) lollipop appearance  produce a positive reagent strip test for protein
.
Germ Tube (-) drumstick appearance

PARASITE
CASTS
Trichomonas vaginalis
 most frequently encountered HYALINE CASTS
 pear-shaped flagellate (motile)  prototype
 manner of reporting: RFMoMa  increased after strenuous exercise, dehydration,
 resembles WBC, transitional or RTE heat exposure, & emotional stress
 pathologically increased in acute
glomerulonephritis, pyelonephritis, chronic renal
disease, & congestive heart failure
Normal Values 0-2 / LPF
STERNHEIMER-MALBIN STAIN
.
 pink color

College Of Medical Laboratory Science │Ranz Barron Bamba

 AUBF311: ANALYSIS OF URINE & BODY FLUIDS
Lecture Module First Semester
Week 05 S.Y. 2023-2024

EPITHELIAL CASTS
 associated with toxicity from heavy metals,
chemicals, or drugs; viral infection & allograft
rejection

RBC CASTS
 more specific, showing bleeding within the
nephron
 associated with damage to the glomerulus FATTY CASTS
(glomerulonephritis)  seen in disorders causing lipiduria
 detected under low power by their orange-red  don’t stain with Sternheimer-Malbin stain
color Sudan III confirmation using polarized
 present in the absence of freestanding RBCs & .
Oil Red O microscopy
positive reagent strip test for blood

WAXY CASTS
 representative of extreme urine stasis, indicating
WBC CASTS chronic renal failure
 signifies infection or inflammation within nephron SUPRAVITAL STAIN
 associated with pyelonephritis & primary marker .
 stain dark pink
for distinguishing pyelonephritis (upper UTI)
from cystitis (lower UTI)
SUPRAVITAL STAIN
.
 demonstrate characteristic nuclei

BROAD CASTS
 referred as renal failure casts
 represent extreme urine stasis
 all types of casts may occurs in broad form
 most commonly seen are granular & waxy
BACTERIAL CASTS
 seen in pyelonephritis
 resembles granular casts
 confirmed by Gram stain on dried or
cytocentrifuged sediment

College Of Medical Laboratory Science │Ranz Barron Bamba

 AUBF311: ANALYSIS OF URINE & BODY FLUIDS
Lecture Module First Semester
Week 05 S.Y. 2023-2024

CRYSTALS
MAJOR CHARACTERISTICS OF NORMAL URINARY CRYSTALS
CRYSTAL pH COLOR APPEARANCE

Yellow-brown (rosette,
Uric Acid Acid
wedges)

Brick dust or yellow

Amorphous Urates Acid
brown

Colorless (envelopes,
Calcium Oxalate Acid/neutral (alkaline)
oval, dumbbell)

Amorphous
Alkaline/neutral White-colorless
Phosphates

Calcium Phosphate Alkaline/neutral Colorless

Triple Phosphate Alkaline Colorless (“coffin lids”)

Yellow-brown (“thorny
Ammonium Biurate Alkaline
apples”

Calcium Carbonate Alkaline Colorless (dumbbells)

College Of Medical Laboratory Science │Ranz Barron Bamba

 AUBF311: ANALYSIS OF URINE & BODY FLUIDS
Lecture Module First Semester
Week 05 S.Y. 2023-2024

MAJOR CHARACTERISTICS OF NORMAL URINARY CRYSTALS

CRYSTAL pH COLOR APPEARANCE

Colorless (hexagonal
Cystine Acid
plates)

Colorless (notched
Cholesterol Acid
plates)

Yellow (concentric
Leucine Acid/neutral
circles)

Colorless-yellow
Tyrosine Acid/neutral
(needles)

Bilirubin Acid Yellow

Sulfonamides Acid/neutral Varied

Radiographic Dye Acid Colorless (flat plates)

Ampicillin Acid/neutral Colorless

College Of Medical Laboratory Science │Ranz Barron Bamba