Urinalysis 4 I. Uses/ Purpose: 1. To detect and identify insoluble materials present in the urine 2.

For the detection of renal or urinary tract disease II. Microscopic Analysis is subject to the following: a. Methods by which the sediment is prepared b. Volume of sediment actually examined c. Methods and equipment used for visualization Page 82 of Strasinger Addis developed the first standardized procedure. Uses hemacytometer to count WBCs and other cells in 12-hour urine specimen. III. A. Standardizing Specimen preparation and volume -freshly voided or adequately preserved preferably first-morning urine -midstream, clean catch specimen -standard volume of urine10-15mL; 12mL is used frequently because the capped centrifuge tubes are calibrated to this volume. B. Time and Speed of Centrifugation -5 min at 1500 RPM oor 400g RCF. To correct for differences in the diameter of centrifuge heads, RCF is used rather than RPM -RCF=1.118 x 105 x radius in cm x RPM2 C. Sediment Preparation Volume of sediment frequently used0.5 to 1.0mL SCF (Sediment Concentration Factor) -detecting elements present in low quantities -used in quantitating the number of elements present per mL of urine Conventional glass slide method1-2 drops or 20uL (0.02mL) Commercial Systems: KOVA (ICL Scientific)capped centrifuge tube Urisystem (Fisher Scientific) Count-10 (V-tech) Quick Read 10 (Globe Scientific) Cen-Slideprovides a specially designed test tube which permits direct reading of microscopy R/S 2000consists of a glass flow cell in which the sediment is found microscopic exam Advantages of Cen slide and R/S 2000 1. closed system 2. does not require manual loading 3. minimize exposure to biohazardous aerosols Advantages of the Commercial System 1. Capped, calibrated centrifuge tubes 2. Decanting pipettes to control sediment volume 3. Slides that control the amount of sediment examined 4. Provide a consistent monolayer of sediment 5. Provide calibrated grids for consistent quantitation

D. Examination of the sediment and reporting of results Get an overview of sediment with LPO. Change to HPO for identification and confirmation especially for RBC and WBC. RBC, WBC= average no./ HPF Casts=average no/ LPF EC, Crystals=uses semiquantitative terms like rare, few, or Plus values Conversion of average # oto # pero mL: E. Microscopic Techniques: The sediments are unstained 1. Bright-field microscopy 2. Phase Contrast Microscopyto enhance visualization of elements with low refractive index (eg. Hyaline cast, mucous threads, Trichomonas, mixed cellular cast) 3. Polarizing Microscopy -aids in the identification of cholesterol in oval fat bodies, fatty casts and crystals 4. Interference-Contrast Microscopy -produces a 3 dimensional image formed and layer by layer imaging of the specimen IV. Sediment Stains Most commonly used stain is the Sterheimer-Malbin stain. Corresponding action and function. You read that one nalang ha? Page 85 table Cytodiagnostic Urinalysis With the aid of cytocentrifuge. Using Papanicolau stain. 1. Calculate the area of an LPF or HPF for the mx used. (pi)r2=area Example: Diameter of HPF=0.35mm 3.14 x 0.1752 = 0.096mm2 2. Calcukation of maximum no of LPF or HPF in viewing area. Area (22 x 22mm) cover slip= 484mm2 484/ 0.096=5040 HPFs V. Calculate the number of HPFs per mL of urine tested using the concentration factor and the volume of sediment explained. 5040