GLOMERULAR DISEASE DIAGNOSIS
In this session of Nephropathology Essentials, Dr. Seltzer provided a comprehensive review of the art and science of urine microscopy. Our Moderator’s Notes are derived from his live presentation


By Dr. Pravir Baxi
Key points:
Review of the urinary sediment can (1) provide insight into the etiology of a patient’s acute kidney injury (AKI) and potentially help differentiate between the various causes, (2) help guide the decision to pursue a kidney biopsy and (3) assess the renal response to therapy
Staining
Sternheimer-Malbin (SM) stain facilitates identification of WBCs, epithelial cells, and casts
Sudan III stain (not used routinely) helps identify lipids and is used an adjunct to polarization
Microscope
X100, x400 and x1000 magnification options are recommended
Condenser: focuses the light on a single point in the plane of view; different illumination modalities are changed via the modification of the light coming from the condenser
Use glass slides and coverslips – not plastic
Verify proper illumination of the specimen – Kohler Illumination (= a process through which the microscope setup is optimized to provide the best visual quality)
Produces uniform bright light which focuses on the specimen
Restricts light exposure of the specimen to the observed field
Illumination modalities – if available, utilize all four
Bright Field
Simplest of all optical microscopy illumination techniques
Darker sample on a bright background
Provides the best resolution in a stained specimen
Dark Field
Excludes the direct light from the image and thus the field around the specimen is dark
Helps illuminate unstained or transparent elements against a dark background
Lower refractive index elements are seen more readily via this modality (such as lipids, crystals, and casts)
Provides lower resolution than the Bright Field modality
Phase Contrast
Enhances contrast of transparent and colorless objects by altering the optical path of light and thus objects will shine out of contrast to adjacent structures
Good for identifying dysmorphic RBCs
Polarized Light
Helps with viewing specimens that are visible primarily due to their optically anisotropic character
Urine Sediment
Cells
Size can help differentiate the type
Cell diameter: Squamous epi cell > transitional epi cell > renal tubular cell > neutrophil > erythrocyte > bacteria
RBCs
Normal RBCs will have a round appearance
Acanthocytes are seen in glomerular hematuria
Not all dysmorphic RBCs are acanthocytes, and while acanthocytes are considered to be relatively specific for glomerular hematuria, dysmorphic RBCs, in general, are rather non-specific and include crenated RBCs, schistocytes, poikilocytes, etc.
RBC mimics that may be visible under the microscope: yeast, air bubbles, starch, calcium oxalate, neutrophils, sperm, fat droplets, and pollen
Neutrophils
Found in infection and/or inflammation
These cells have a granular cytoplasm and a segmented multi-lobed nucleus
The SM stain facilitates visualization of the segmented nucleus
Glitter Cells – pale staining WBCs with granular motility but these are a non-specific finding
Lipids
Oval fat bodies are renal tubular epithelial cells containing fat globules
They are formed when lipids coalesce into refractile globules on absorption by renal tubular cells
These oval fat bodies can also be found within a cast
Sudan III stains are helpful in that it colors the lipids orange
Casts
Formed via solidification of Tamm-Horsfall mucoprotein with other cells/structures within the renal tubules
Hyaline casts: most common, not pathologic, normal in concentrated urine
Granular casts
Can result from the breakdown of cellular casts or degenerative products of tubular cells and proteins
Classified as fine or coarse, but this has no diagnostic significance
These are indicative of acute tubular necrosis (ATN)
Pigmented casts: hemoglobin, myoglobin, bilirubin, and drug pigments
WBC casts
Indicative of inflammation or infection
WBC casts can be present in patients with proliferative glomerulonephritis (GN)
RBC Casts
Signify the presence of a proliferative GN or vasculitis process
Renal Tubular Epithelial casts: indicative of ATN
Waxy casts: seen in chronic renal failure and thought to represent the end product of cast revolution
Lipid casts: seen in nephrotic syndrome
Pseudo-casts: cylindrical appearing structures that resemble a true cast
Formed when cells or particles adhere to a mucous thread
Crystals
Artifacts
Selected References: Fogazzi, G. B., Ponticelli, C., & Ritz, E. (1999). The Urinary Sediment: An Integrated View. Oxford University Press. Haber, M. H., Blomber, D., Galagan, K., Glassy, E. F., & Ward, P. C. (2010). Color Atlas of the Urinary Sediment: An Illustrated Field Guide Based on Proficiency Testing. College of American Pathologists. Seltzer, J., Velez, J. C., Buchkremer, F., & Tesser, J. A. (n.d.). Renal Fellow Network (RFN) . Retrieved from https://www.renalfellow.org/category/urine-sediment-of-the-month/