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Microscopic Exam

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Microscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of UrineMicroscopic Examination of Urine Download http://www.vetlab.com/kova.htm Definition of urine sediment: all solid materials suspended in the urine - a semiquantative evaluation of the urine sediment Significance of formed elements in the urine Well performed microscopic exam can provide information nearly equivalent to a biopsy. Most time consuming part of UA & until recently the least standardized. Ongoing controversy as to when / if to perform the microscopic exam.Microscopic Examination of Urine Not on lecture guide. Review info in Table 6-1 Correlation of findings from physical & chemical analysis with expectations in microscopic.Screening Test Significance (or what to look for)Nitrite positive WBCs / bacteriaLeukocyte esterase pos WBCs, WBC casts, bacteriaGlucose positive yeastsMicroscopic Examination of Urine Specimen requirements Collection of specimen Prefer the concentrated first morning specimen, collected = mid-stream, clean catch . first morning most concentrated and will be able to demonstrate the most abnormalities. Mid stream, clean catch technique will eliminate fecal & vaginal contamination  Container must be clean and free of lint / debris usually disposable plastic, must be sure no soap residue Fresh – tested within 2 hours of voiding, or refrigeration needed.Microscopic Examination of Urine Obj.35. List the correct steps in the collection and preparation of a urine sample for microscopic exam. Preparation of specimen need to standardize as much as possible Sources of Variation (not on lecture guide) Collection method Centrifugation time and speed Re-suspension of sediment Type of microscope slide Viscosity of specimen Reporting of the resultsMicroscopic Examination of Urine Preparation of specimen (show video) Mix specimen well Pour 12 ml into urine centrifuge tube Centrifuge five minutes, 1200-2000 RPM (speed varies depending on the centrifuge’s characteristics) Speed and time should be consistent. The “relative centrifugal force” is important.Microscopic Examination of UrineMicroscopic Examination of Urine Pour off supernatant - except last .5-1 mL. have pipettes that assist Re-suspend sediment - mix well, tap, or use pipette provided Evaluate sediment in a chamber standardized for given volume and depth of field. - “In-house methods = Mount a smalldrop on a clean slide, cover-slip - or use commercial materials such as Count 10 Use standardized reporting format consistent with other techs in the institutionMicroscopic Examination of Urine Commercial systems UriSystem – slide to follow KOVA System – video or several slides to follow Count -6 or Count 10 all have their ‘own brand’ of tubes, pipettes, stain, slides, etc. Authors also mentions several other ‘all in one-type of systems’Microscopic Examination of Urine UniSystem Standardization of Urine SedimentMicroscopic Examination of Urine Sedi-Stain (Sternheimer and Malbin) crystal violet, safranin-O Sedi-Stain & KOVA stain are commercial preparations with addition of stabilizers to prevent precipitation.Supra-vital stain used to increase visibility of structures. Assists greatly in differentiating renal tubular epithelial cells (which will take on an eosinophilic - oranges cytoplasm & dk purple nuclei) from transitional epithelial (which are more over-all blue)Microscopic Examination of Urine Not on lecture guide – Table 6-3 Sediment stain characteristics Toluidine blue – nuclear structure Assists in differentiating WBC from renal epith. 2% acetic acid - removes interfering RBCsand enhances nuclei of WBC Lipid stains - Oil Red O, Sudan III - stains triglycerides and neutral fats orange-red to ID lipid containing cells.Microscopic Examination of Urine Gram stain - to assist in ID of gram reaction of bacteria. Hansel stain - methylene blue and eosin Y stains eosinophilic granules - ID eosinophils Prussian blue reaction - makes iron granules blue in color (hemosiderin granules appear yellow until stained)Microscopic Examination of Urine Table 6-5 – page 73 provides information on types of microscopic techniques that have application in UABrighfield microscope – very subdued light: lowered condenser, closed iris diaphragm, use filters Continuously focus up and down with fine adjustment as you learned in hematology. Polarized light - may use to ID crystals, lipidsMicroscopic Examination of Urine Types of Sediment As one author puts it: Cells Casts Crystals CrittersMicroscopic Examination of Urine Types of Sediment Organized – biological part RBC WBC Casts Epithelial cells Bacteria, parasites, yeast and fungi Unorganized Crystals Amorphous crystalline matter.Microscopic Examination of Urine Examination - should correlate with physical and chemical dipstick, may need to recheckScanning - – 10-15 fields using low power (10X). Look for casts, mucous, and squamous epithelial cells in general getting an overall feel Report squamous epithelial cells, crystals, mucous, etc. using semi-quantitative terms such as rare, few, moderate, or many (or trace, 1+,2+,3+, & 4+) according to lab protocol.Microscopic Examination of Urine Enumeration - quantitate. Method may vary from lab to lab Average number of RBC/hpf Average number of WBC/hpf Average number of any renal tubular or transitional epithelial cells /hpf.Microscopic Examination of UrineAverage number (and type) of casts/__average # of casts /hpf______ authors have varied back and forth as whether low or high power should be reported... use low power to locate and enumerate the various types , but may need to switch to high power


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