Microscopic Examination Of Urine Flashcards by vnlla babe

Urine Sediment Preparation

10-15 mL urine ( average: 12 mL)

Centrifuge for 5 minutes @ 400 RCF

Decant urine (0.5 or 1.0 mL remains)

Transfer 20uL (0.02 mL) sediment to glass slide with 22x22 mm coverslip

Examine microscopically

10 LPF, 10 HPF under subdued light (reduced light/dim light)

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Quantitative measure of formed elements of urine using hematocytometer

Addis Count

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Addis Count specimen

12 hours specimen

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Addis count preservative

Formalin

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Addis Count normal values: RBCs

0-500,000/12hr urine

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Addis Count normal values: WBCs

0-1,800,000/ 12hr urine

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Addis Count normal values: Hyaline Casts:

0-5,000/12hr urine

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Used for routine Urinalysis

Bright-field microscopy

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Enhance visualization of elements with low refractive index (ex: hyaline cast)

Phase-contrast microscopy

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Identification of cholesterol in oval fat bodies, fatty casts and crystals

Polarizing microscopy

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Identification of Treponema pallidum

Dark-field microscopy

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Visualization of fluorescent microoganisms or those stained by a fluorescent dye

Fluorescence microscopy

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-used for 3D imaging & layer-by-layer imaging of specimen

Interference contrast microscopy

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2 types of Interference contrast microscopy

a. Nomarski (Differential)
b. Hoffman (Modulation)

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-adapted in Bright field microscopy

a. Nomarski (Differential)
b. Hoffman (Modulation)

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Delineates structure and contrasting colors of the nucleus and cytoplasm

Identifies WBCs, epithelial cells, and casts

Sternheimer-Malbin

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Enhances nuclear detail

Differentiates WBCs from RTE cells

Toluidine blue

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Lyses RBCs and enhances nuclei of WBCs

Distinguishes RBCs from WBCs, yeast, oil droplets, and crystals

2% acetic acid

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Stain triglycerides and neutral fats orange-red; do not stain cholesterol

Identify free fat droplets and lipid-containing cells and casts

Lipid stains: Oil Red O and Sudan III

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Differentiates gram-positive and gram- negative bacteria

Identifies bacterial casts

Gram stain

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Methylene blue and eosin Y stains eosinophilic granules

Identifies urinary eosinophils

Hansel stain

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Stains structures containing iron

Identifies yellow-brown granules of hemosiderin in cells and casts

Prussian blue stain

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Combination stain for Sternheimer

crystal violet + safranin o

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Combination stain for Hansel

Methylene blue + eosin y

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stains DNA

Phenathridine (orange)

stains nuclear membranes, mitochondria and cell membranes

Carbocyanine (green)

Sediment Constituents

RBCs (Hematuria) WBCs (Pyuria or Leukocyturia) Epithelial Cells Bacteria YEASTS PARASITES Spermatozoa Mucus threads

Smooth, non-nucleated, biconcave disks (HALO)

RBCs (Hematuria)

RBCs (Hematuria) Normal value:

0-2, 0-3/HPF

RBCs in Hypertonic solution

crenated/ shrink

RBCs in Hypotonic solution

swell/burst , hemolyze (Ghost cells)

RBCs in Glomerular membrane damage:

RBC look dysmorphic w/ projections and fragmented

RBCs (Hematuria) Sources of Error (4)

Calcium oxalate, oil droplets, yeast, air bubbles

RBCs (Hematuria) Remedy

add 2% acetic acid. It will lyse the RBCs but not the others

presence of WBC in urine

Pyuria

WBCs Normal value:

0-5 or 0-8/ HPF

Increase number of WBCs indicates the presence of

infection or inflammation

Predominant WBC in urine

Neutrophils

Neutrophils in Hypotonic urine they swell & the granules undergo ________ movement which produce ________ appearance=________ cells

Brownian; sparkling; Glitter

Eosinophils normal value

<1%

Eosinophils significant value?; increase eosinophils assoc. w/ ________

>1%; acute interstitial nephritis

monocytes, lymphocytes, macrophages, histiocytes Present in small numbers

Mononuclear cells

REPORTING of RBCs & WBCs; Numerical ranges:

10 fields per HPF 0-2, 2-5, 5-10, 10-25, 25-50, 50-100, >100

point of reference when focusing the microscope

Squamous epithelial cells

LARGEST CELLS; From lining of vagina, female urethra and lower portion of male urethra

Squamous epithelial cells

Irregular cytoplasm and prominent nucleus

Squamous epithelial cells

squamous epithelial cells covered with Gardnerella vaginalis which causes bacterial vaginosis

CLUE CELLS

CLUE CELLS- squamous epithelial cells covered with _________ which causes bacterial vaginosis

Gardnerella vaginalis

Transitional epithelial cell is also known as (2)

Urothelial Cells or Bladder Cells

Spherical, polyhedral, caudate w/ centrally located nucleus

Transitional epithelial cell

Derived from the linings of renal pelvis, ureter, urinary bladder and upper portion of male urethra

Transitional epithelial cell

High T.E.C came from Catheterization procedure. abnormal morphology may indicate _____ or _____.

malignancy or viral infection

T.E.C Can be seen in

in pairs, or in clumps (syncytia)

Most clinically significant epithelial cells

Renal tubular epithelial (RTE) cells

Rectangular, polyhedral, cuboidal or columnar, eccentric nucleus

Renal tubular epithelial (RTE) cells

_______ RTE/HPF􏰀 indicates Tubular injury (ex. Glomerulonephritis, acute tubular necrosis, etc.)

>2 RTE/HPF

Larger than leukocytes (WBCs)

Renal tubular epithelial (RTE) cells

lipid containing RTE 􏰀 seen in Lipiduria (ex. Nephrotic syndrome) 􏰀 Identified by: • Lipid stains (TG and neutral fats) • Polarizing microscope (+) Maltese cross appearance

Oval Fat Bodies

Oval Fat Bodies are identified by? (2)

• Lipid stains (TG and neutral fats) • Polarizing microscope (+) Maltese cross appearance

RTE with non-lipid filled vacuoles Seen in ACUTE TUBULAR NECROSIS

Bubble Cell

Reporting of Epithelial Cells Rare, Few, Moderate, Many •Squamous (_____) • Transitional (_____) Average number per HPF (2)

0-5 5-20 20-100 >100 LPF HPF • RTE cells • Oval fat body

presence of WBC and Bacteria

UTI

If presence of bacteria only-contamination, it is an?

old specimen

most common cause of UTI

Enterobacteriaceae (ex. E. coli) Staphylococcus, Enterococcus

Bacteria Reporting: Rare, few, moderate, many /HPF

0-10 10-50 50-200 >200

True yeast cell infection= _____ + _____

WBC + Yeast

If presence of yeast cells only-

contamination

Small, refractile oval structures that may or may not bud

YEASTS

seen in DM & vaginal moniliasis

Candida albicans

YEASTS Reporting:

Do not quantitate but only note the presence. Yeast cells seen / budding yeast cells/ mycelial form/ budding yeast cells with hyphae

most common parasites in urine

Trichomonas vaginalis

”Ping pong disease”

Trichomonas vaginalis

Pear-shaped flagellate with jerky motility

Trichomonas vaginalis

PARASITES • Trichomonas vaginalis reporting

Reporting: Rare, few, moderate, many/ HPF ***Do not quantitate but only note the presence.

Blood fluke with terminal spine Causes hematuria and cancer in the bladde

Schistosoma haematobium ova

Schistosoma haematobium ova Specimen

24 hours unpreserved urine

Schistosoma haematobium ova Bladder cancer marker: (2)

• NMP (nuclear matrix protein) • BTA (Bladder Tumor Antigen)

Most common fecal contaminant

Enterobius vermicularis ova

After sexual intercourse Reporting: Present ***Do not quantitate but only note the presence

Spermatozoa

Mucus threads Major constituent:

TAMM-HORSFALL PROTEIN (uromodulin)

Mucus threads Reporting: Rare, few, moderate, many/ LPF

0-1 1-3 3-10 >10