Microscopic Examination Of Urine Flashcards
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)
Quantitative measure of formed elements of urine using hematocytometer
Addis Count
Addis Count specimen
12 hours specimen
Addis count preservative
Formalin
Addis Count normal values: RBCs
0-500,000/12hr urine
Addis Count normal values: WBCs
0-1,800,000/ 12hr urine
Addis Count normal values: Hyaline Casts:
0-5,000/12hr urine
Used for routine Urinalysis
Bright-field microscopy
Enhance visualization of elements with low refractive index (ex: hyaline cast)
Phase-contrast microscopy
Identification of cholesterol in oval fat bodies, fatty casts and crystals
Polarizing microscopy
Identification of Treponema pallidum
Dark-field microscopy
Visualization of fluorescent microoganisms or those stained by a fluorescent dye
Fluorescence microscopy
-used for 3D imaging & layer-by-layer imaging of specimen
Interference contrast microscopy
2 types of Interference contrast microscopy
a. Nomarski (Differential)
b. Hoffman (Modulation)
-adapted in Bright field microscopy
a. Nomarski (Differential)
b. Hoffman (Modulation)
Delineates structure and contrasting colors of the nucleus and cytoplasm
Identifies WBCs, epithelial cells, and casts
Sternheimer-Malbin
Enhances nuclear detail
Differentiates WBCs from RTE cells
Toluidine blue
Lyses RBCs and enhances nuclei of WBCs
Distinguishes RBCs from WBCs, yeast, oil droplets, and crystals
2% acetic acid
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
Differentiates gram-positive and gram- negative bacteria
Identifies bacterial casts
Gram stain
Methylene blue and eosin Y stains eosinophilic granules
Identifies urinary eosinophils
Hansel stain
Stains structures containing iron
Identifies yellow-brown granules of hemosiderin in cells and casts
Prussian blue stain
Combination stain for Sternheimer
crystal violet + safranin o
Combination stain for Hansel
Methylene blue + eosin y
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