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“Urine-Titled” to Know: Urine Sediment Evaluation

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“Urine-Titled” to Know: Urine Sediment Evaluation Elaine Anthony, MA, CVT TECH St. Petersburg College TALK Largo, Florida “Urine-Titled” to Know: Urine Sediment Evaluation The primary purpose of microscopic examination of urine sediment is are consistent when obtaining and to detect abnormal formed elements (eg, cells, casts, crystals) in the evaluating the urine than the amount of urine obtained. Every practice sample. Urine sediment may contain a variety of miscellaneous compo- should clearly communicate its proto- nents, some that have clinical significance (eg, parasites, bacteria), and col and standards to ensure accuracy. some that are strictly contaminants (eg, starch granules, pollen, grass The urine sample should be centri- particles). The presence or absence of specific formed elements can fuged at a low speed (eg, 1500–2000 RPM) for 5 minutes. The supernatant provide detailed diagnostic information. is then decanted and the sediment re- suspended in the remaining urine. A Urine sediment examination should utilize 5 mL of fresh urine.1 Normal refer- drop of this suspension is placed on a ence ranges are based on a particular urine amount and any alteration of the glass slide (Figure 1) with a cover slip amount should be considered. However, it is more important that technicians and examined microscopically. 10 veterinaryteambrief.com April 2014 PEER REVIEWED Unstained Urine Sediment Overall cellularity can be assessed, To evaluate the unstained urine sedi- and bacteria, parasites, and other in- ment slide, first scan the slide with a fectious agents can be seen. Next, pro- low-power objective lens (10×) and ceed to the high-power (40×) objective subdued lighting (iris diaphragm par- lens while slightly increasing the light tially closed and condenser in the low- and raising the condenser. ered position).1,2 To ensure the viewing level is actually urine sediment and Stained Urine Sediment not the cover slip or slide, find and Supravital Sediment Photo by Vivian Tiffany, BA, CVT 2 Figure 1. Placing a drop of suspension on a glass focus on any cellular component. A wet preparation may be desired as slide Once this is established, perform a a follow-up to enhance the refractive scan on low power. Large formed ele- index for examination (see Wet- and ments (eg, casts, crystals) or clumps of Dry-Preparation Stains). One ad- cells are evident at 10× magnification. vantage of the supravital stain is that Wet- and Dry-Preparation Stains Cells can be difficult to identify or differentiate using unstained urine sediment analysis because swollen white blood cells may be confused with renal Figure 2. Diff-Quik slide preparation materials epithelial cells, or contaminant material may be confused with formed elements. Clusters of transitional cells require further examination on oil immersion to determine if they are normal, reactive, or neoplastic via evaluation of the stained urine sediment: • Wet-preparation supravital stain (Sedi-Stain [bd.com]; new methylene blue): Mix a drop of stain with the suspended sediment and incubate at room temperature for 2–3 minutes before placing a drop of stained sediment onto a microscope slide. Figure 3. Wright-Giemsa stain slide preparation • Dry-preparation Romanowsky stain (Diff-Quik [Figure 2]; Wright-Giemsa materials stain [Figure 3]): Place one drop of suspended sediment on a clean micro- scope slide. Using a second clean microscope slide, make a compression smear (Figure 4). Air dry the top slide thoroughly and stain gently. TIP Because of the low-protein nature of urine specimens, cellular components of urine sediment may not adhere to the slide.1 Agitation in the staining process can also result in loss of the sediment from the slide. Increasing the fixative time will improve the cell’s adherence; more than 1–2 minutes of fixative time for urine cytology is recommended.3 Stain as gently as possible, considering the rinse of a gentle flow of water. Staining time may be 1.5–2 times the amount used for hematology slides, depending on the cellularity of the sediment. The slide should Photo by Vivian Tiffany, BA, CVT then be air dried thoroughly and evaluated on low-, high-, and oil-immersion. Figure 4. Creation of a compression smear sample April 2014 Veterinary Team Brief 11 TECH TALK Her hunt for effective To be of greatest value, efforts must be made to standardize the flea control performance and reporting of results.2.27 x 9.7 Brief Team Veterinary is over it does not contain a fixative, which to peripheral blood—red and white dissolves crystals. Counts must be blood cells are easily identified. performed on the unstained sediment From this vantage point, other ele- slide first, as quantifying the sedi- ments can be compared with cells ment’s elements once diluted with the that are now easily identified (eg, stain yields inaccurate results. blood cells vs transitional epithelial cells). With wet-preparation evalua- Problems may result from a supravi- tion, white blood cells become more tal stain that can actually make the difficult to identify when swollen, identification more complicated; for and bacteria can be challenging to Account Coordinator: example, if bacteria and stain precip- detect when found in groups COM031099P560CVZ or itant accumulate over time and are when contaminants are Account Service: involved. not filtered, they can be uninten- The dry-preparation cytology date: Today’s stain Art Director: And so is yours tionally added to the urine sediment. procedure can helpProduction: differentiate these Proofing: In addition, the stain will occasion- aspects and confirm what was seen on That’s because you already have our ally stain cytoplasm the same color direct sediment evaluation. If a cluster fast-acting, month-long as the nucleus, making it difficult to of suspicious transitional epithelial flea control tablet for 02-04-14 10:39 AM clearly differentiate them. To see nu- cells is encountered, further evaluaDue: 3-14-14 - cats on your shelf. clear and cytoplasmic detail, particu- tion under oil immersion is needed for Call off the search larly when identifying nuclear criteria nuclear criteria of malignancy.2 and reach for the of malignancy, oil immersion is need- easy prevention and Conclusion treatment recommendation ed, which the supravital wet prepara- you know and trust. tion does not allow. Microscopic examination of urine sediment is a rapid, easy-to-perform Romanowsky Sediment diagnostic procedure that provides comfortis.com When cellular elements are difficult valuable information. To be of great- 888-545-5973 to identify, the preferred (and simple) est value, efforts must be made to method is to make a dry-preparation standardize the performance and re- or air-dried cytology slide (see Wet- porting of results. Unstained and and Dry-Preparation Stains, pre- air-dried cytology can enhance the vious page). The slide is stained with amount of information for reliable Important Safety Information a Romanowsky-type stain, such as results. For cats: The most common adverse reaction recorded in clinical trials was vomiting. Other adverse reactions Diff-Quik. There is comfort in view- were lethargy, anorexia, weight loss, and diarrhea. Use with caution with concomitant extra-label use of ivermectin. ing an air-dried cytology slide on oil See Aids & Resources, back page, For product label, including complete safety information, see page 12. immersion because of its correlation for references & suggested reading. ©2014 Elanco CF00999 12 veterinaryteambrief.com April 2014 Veterinary Team Brief COM031099P560AVA Due: 3-14-14 8.125 x 10.875 Today’s date: 02-04-14 10:40 AM Account Service: Account Coordinator: Art Director: Production: Proofing:.
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