1030 Unit 3 Flashcards
macroscopic screening of urine specimen is used to what?
increase cost-effectiveness of urinalysis
variations in the microscopic analysis of urine include what?
–preparation of urine sediment
–amount of sediment analyzed
–method of reporting
what can cause false negative microscopic results?
–braking the centrifuge
–failing to mix the specimen
–diluting alkaline urine
what are the two factors that determine relative centrifugal force?
–diameter of rotator head
–RPM
when using the glass-slide and cover-slip method, what might be missed if the coverslip is overflowed?
casts
initial screening of the urine sediment is performed using what objective power?
10x
what should be used to reduce light intensity in bright-field microscopy?
rheostat
which of the following are reported as number per lpf?
A. RBCs
B. WBCs
C. Crystals
D. Casts
casts
The Sternheimer-Malbin stain is added to urine sediments to do what?
–increase visibility of sediment constituents
–change the constituents refractive index
–delineate constituents structures
Nuclear can be enhanced by what?
–toluidine blue
–acetic acid
what lipids are stained by sudan III?
–Neutral fats
–triglycerides
what lipids are capable of polarizing light?
cholesterol
What is the Hansel stain identify?
eosinophils
crenated RBCs are seen in urine that is what?
hypersythenuric
Differentiation among RBCs, yeast, and oil droplets may be accomplished by what?
lysis of yeast cells by acetic acid
what does dysmorphic RBCs indicate?
glomerular bleeding
leukocytes that stain pale blue with Sernheimer-Malbin stain and exhibit brownian movement are what?
glitter cells
sometimes mononuclear leukocytes are mistaken for what?
RTE cells
when pyuria is detected in a urine sediment, the slide should be checked for the presence of what?
bacteria
transitional epithelial cells are sloughed from the what?
bladder
the largest cells in urine sediment are what?
squamous epithelial cells
what make a squamous epithelial cell clinical significant?
clue cell
forms of transitional epithelial cells include what?
convoluted
increased transitional cells indicate what?
–catheterization
–malignancy
what is a primary characteristic used to identify renal tubular epithelial cells?
eccentrically located nucleated
After an episode of hemoglobinuria, RTE cells may contain what?
hemosiderin granules
the predecessor of the oval fat body is what?
Renal tubular cell (RTE cell)
a structure believed to be an oval fat body produced a Maltese cross formation under polarized light but does not stain with Sudan III. What is this structure?
cholesterol
the finding of yeast cells in the urine is commonly associated with what?
diabetes mellitus
what is the primary component of urinary mucus?
uromodulin
where are majority of casts formed?
DCT
cylindruria refers to the presence of what?
all type of casts
a person submitting a urine specimen after a strenuous exercise routine normally can have what components?
WBC casts
before identifying an RBC cast, what should be observed?
–free-floating RBCs
–intact RBCs in the cast matrix
–a positive reagent strip blood reaction
what are WBC primarily associated with?
pyelonephritis
what is the shape of RTE cells associated with RTE casts primarily?
round
when observing RTE casts, the cells are primarily what?
attached to the surface of a matrix
the presence of fatty casts is associated with what?
–nephrotic syndrome
–crush injuries
–diabetes mellitus
what do nonpathogenic granular casts contain?
cellular lysosomes
what are some characteristics of waxy casts?
–represent extreme urine stasis
–may have a brittle consistency
–contain degenerated granules
what does observation of broad casts represents what?
–destruction of tubular walls
–formation in the collecting ducts
what contributes to urinary crystal formations?
–pH
–solute concentration
–temperature
the most valuable initial aid for identifying crystals in a urine specimen is what?
pH
what are crystals associated with severe liver disease?
–tyrosine
–bilirubin
–leucine
what are some crystals that routinely polarize?
cystine
casts and fibers usually can be differentiated using what?
polarized light
what crystal is described as “envelopes” in acidic urine?
calcium oxalate dihydrate
what crystal is described as “yellow-brown; whetstone” in acidic urine?
uric acid
what crystal is described as “pink sediment in acidic urine?”
amorphous urates
what crystal is described as “ovoid” in acidic urine?
calcium oxalate monohydrate
what crystal is described as “coffin lids” in alkaline urine?
triple phosphate
what crystal is described as “ white precipitate” in alkaline urine?
amorphous phosphate
what crystal is describe as “thin prisms” in alkaline urine?
calcium phosphate
what crystal is described as “thorny apples” in alkaline urine?
ammonium biurate
what crystal is described as “dumbbell shape” in alkaline urine?
calcium carbonate
what abnormal crystal is described as “ hexagonal plates”?
cystine
what abnormal crystal is described as “fine needles seen in liver disease”?
tyrosine
what abnormal crystal is described as “notched corners”?
cholesterol
what abnormal crystal is described as “concentric circles, radial striations”?
leucine
what abnormal crystal is described as “bundles after refrigeration”?
ampicillin
what abnormal crystal is described as “flat plates; high specificity gravity”?
radiographic dye
what abnormal crystal is described as “bright yellow clumps”?
bilirubin
what type of microscope would be described as “low-refractive-index objects may be overlooked”?
bright field
what type of microscope would be described as “forms halo of light around object”?
phase
what type of microscope would be described as “object split light into tow beams”?
polarized
what type of microscope would be described as “indirect light is reflected off the object”?
dark-field
what type of microscope would be described as “detect specific wavelengths of light emitted from objects”?
fluorescent
what type of microscope would be described as “3D images”?
interference contrast
list physical and chemical parameters included in microscopic urine screening?
–color
–clarity
–blood
–nitrite
–protein
–leukocyte esterase
–glucose
what is the significance of color in microscopic urine screenig?
RBC
what is the significance of clarity in microscopic urine screening?
–hematuria vs. hemoglobinuria/myoglobinuria
–confirm pathological and non pathological cause of turbidity
what is the significance of blood in microscopic urine screening?
RBC and RBC cast
what is the significance of nitrite in microscopic urine screening?
bacteria and WBC
what is the significance of protein in microscopic urine screening?
casts and cells
what is the significance of leukocyte esterase in microscope urine screening?
WBC, WBC casts, bacteria, and yeast
what is the significance of glucose in microscopic urine screening?
yeast
what is an advantage of commercial systems over glass-slide method for sediment?
provides a tube is specially designed to permit direct reading of urine sediment
Microscopic analysis is subject to what procedural variations?
—well mixed urine is essential
—how the sediment is prepared
—volume of sediment to examine
—method and instrumentation used to visualize the sediment or elements
—the nomenclature is which the results are reported
What are requirements for specimen preparation of fresh/preserved specimen?
— RBC And WBC cells, casts disintegrate in dilute alkaline urine
—has precipitation after refrigeration, amorphous urates and amorphous phosphates
—warm specimen to 37 C before centrifugation
—type of collection may prevent contamination (mid stream)
What are requirements of specimen preparation for the volume?
—standard: 10-15 ml
—quantities <12 ml should be documented (too small will reduce formed element amount
What are the requirements for specimen preparation of centrifugation?
—speed and length of time
—speed 2 ways to measure centrifugation: relative centrifugal force (RCF) and revolutions per minute (RPM)
—RCF = 1.118 x 10^ -5 x radius in cm x RPM^2
—5 minutes
What are the requirements for specimen preparation of Sediment preparations?
—uniform amount should remain after decanted = 1.5 - 1.0 ml
—after decanting, the sediment may be resuspended using a puppet. NOT vigorous because it will result in destruction of cellular content
Describe sediment examination
—could use commercial system
—needs to be consistent each time, each tech
— in pipet, place one drop of urine on to glass slide. Place cover slip on top of urine
—do not overflow
— examine 10 fields under low and high power
What are some examples of a commercial urine microscopic system?
—Kova
— urisystem
— count-10
— quick-prep
What is reported on lpf?
Casts
What is reported on hpf?
—epithelial cells
—crystals
— RBC and WBC
What is a stain?
Increases visibility of the elements
What is a supravital stain? And are examples?
—contrasting color between the nucleus and the cytoplasm
—Sternheimer-Malbin:helps visibility of epithelial cells, WBCs, cast
—toluidine blue: helps distinguish between WBC and RTE cells p
What are lipid stains and what are examples?
— triglyceride, neutral fats and cholesterol. Stain orange-red
—oil red O
—Sudan III
What is Hansel stain and give examples?
—helps distinguish eosinophils from WBC
-renal transplant
-pyelonephritis
-allergic reaction
-dry fixed slide
What is Prussian and blue and what are examples?
Stains hemosiderin (iron)
What is acetic acid?
Helps with nuclear detail. Can not use initial sediments because it will lysis the RBCs
What does cytodiagnostic aid in
Cytology
What specimens would be referred to cytodiagnostics?
— detection for malignancies of lower urinary tract
—provides more definite information about renal tubular changes associated with transplant: viral, fungal are parasitic infections, cellular inclusions, pathologists casts, and inflammatory conditions
What stain is used for cytodiagnostic
Papincolaou stain
Describe bright-field microscopy
—used for routine analysis
—objects appear darker against light background
—do not use in high light
Describe phase-contrast microscopy
—enhances visualization of elements with low refractive indices (hyaline cast, mixed cellular cast, mucous, and trichimonas
— as light ray pass an object, they are slowed in comparison to rays passing through air, thereby decreasing the intensity of light and producing contrast. This is called pause difference and is affected by thickness of object
Describe a polarizing microscope?
—aids in identification of cholesterol in oval fat bodies, fatty cast and crystals
—lipids and crystals have the ability to rotate the path of undirectional polarized beam to produce characteristic colors in crystals and Maltese cross in lipids
Describe dark-field micrscope
—aides in identification of Treponema pallidum
—used to enhance specimen visualization that can not easily be seen
—field of view is black. Light refracts/refracts off specimen and is captured by objective lens
Describe fluorescence microscope
—-allow naturally fluorescent microorganisms or those stained by fluorescent, including tagged antigens or antibodies (indirect fluorescent antibodies IFA)
—immunofluorescence method
Describe interference-contrast
—produces a 3D image and layer by layer
—shows very fine structure structure details by splitting the light ray so that the beams pass through different areas
Describe electron microscope
Used for investigation of ultrastructure in biological and inorganic specimens
What are the parts of the Lens system on a microscope?
—oculars
—objectives
—adjustment knows (fine and coarse)
What are the parts of the illumination system on a microscope?
—light source
—condenser
— field
—iris diaphragm
What are the parts of the Body on the microscope?
—base
—body tube
—nose piece
—mechanical stage
What is the information printed on the side of each objective lens ?
—type of objective (4x, 10x, 40x)
— magnification (0.1, 0.25, 0.65)
—numerical aperature
—tube length
—cover slip thickness to be used
What is numerical aperture?
—Refractive Index in material needed between the slide and the outer lens (air or oil).
—higher the aperture, the better the light yielding better resolution
What does the diaphragm control?
The diameter of the light beam
What does the condenser do?
Controls the uniformity of the light. Condenser up and down
What should not be used to regulate the light intensity?
The aperture diaphragm
What are normal urine sediments
—epithelial cells
—RBC
—WBC
—hyaline cast
What could change normal urine sediment?
—concentration
—pH
—presence of metabolites
Describe an isotonic solution
—has smooth non-nucleated RBC
—biconcave disk
Describe hypersthenuric solution
—cause the RBC to shrink due to loss of water from being in a concentrated urine —> creanted
Describe hyposthenuric solution
—Caused from being in dilute urine
— RBC take in too much water, swell and lysis —> ghost cells
What are clinical significance for RBC?
—normal range: 0-2/ hpf
—indicates a glomerular problem either with membrane damage or vascular injury to genitourinary tract
— the number is indicative of the.l damage
—hematuria
—glomerulonephritis, toxic/immunologic reaction, menstrual cycle
Describe macroscopic hematuria
—cloudy, red to brown > 100/lpf
—advance glomerular damage
—vascular integrity of urinary tract due to trauma, infection or inflammation or coagulation disorders
Describe microscopic hematites
—clear urine with RBCs present
— is helpful for early detection
—glomerular disorders
—malignancy of urinary tract
—presence of renal calculi
What is the predominant WBC found in urine?
Neutrophils
Describe neutrophils found in urine
—they contain granules and multi-loved nucleus
—in alkaline urine —> neutrophils lose their nuclear detail
What WBC are abnormal to find?
—eosinophils: seen in drug induced interstitial nephritis, a few in UTI, and renal transplant. 1% is significant because not normally found in urine
— mononuclear cells: lymphocytes, monocytes, macrophages, histocytes. Early stage renal transplant rejection
What are some characteristics of WBC and urine?
—migrate from tissue to site if infection/inflammation
—Pyuria indicate infection or inflammation
—WBC + bacteria = important!!!
—normal range = 0-5/hpf
What are bacterial infections of WBC?
—pyelonephritis
—cystitis
—prostatitis
—urethritis
What are non bacterial infections of WBC?
—glomerulonephritis
—lupus
—interstitial nephritis
—tumors
What is the most common cell to be seen because they are the lining cells of the genitourinary system?
Epithelial cells. They slough off with age
What are the 3 types of epithelial cells?
—squamous
—transitional
—RTE
What are some characteristics of squamous epithelial cells?
—largest cell seen in urine
—abundance of irregular cytoplasm
—report as numerical range from hpf
—originate from lining of vagina, male/female urethra.
—mid stream clean catch reduces squamous cell count
—clue cells (cell surface must be covered 70% in bacteria to be considered this)
What is a clue cell?
Squamous cell with bacteria in the cell which is an indication of vaginal infection with Gardenerella vaginalis
Describe transitional epithelial cells
— several forms:
——> spherical - direct contact with water in bladder become large and round
——> polyhedral - multiple sides
——> caudate - appear to have a tail
— distinct nucleus that is centered
—originates I’m lining of renal pelvis, calyces, ureters, and bladder. Male upper urethra
—increase after catheterization
—if it has vacuoles and irregular nuclei: may indicate malignancy or viral infection
Describe RTE Cells that originate in the PCT
—larger than other RTE cells
—rectangular or columnar or convoluted cell
— cytoplasm has coarse granules with a nucleus
Describe RTE cell that originates in the DCT?
—round or oval
—nucleus is eccentrically
Describe RTE cells that originate from collecting duct
—cuboidal (NEVER ROUND)
—eccentrically nucleus
—one straight edge
—appear in groups of three or more = renal fragment
—can be seen in sheets
Describe an RTE Cell
—report in defined ranges under hpf
— >2 RTE = tubular injury
— most significant epithelial cell
—can take in bilirubin, hemosiderin (Prussian blue)
When can abnormally high RTE cell count be seen?
—necrosis of the renal tubules
——>exposure to heavy metals
——>drug induced toxicity, salicylate poising
——> viral infection (Hep. b)
——> pyelonephritis
——> malignancies
——> allergic reaction or transplant rejection
Describe oval fat bodies
— RTE that have absorbed lipids
— dry refractive. Hard to see nucleus
—usually seen with free floating fat
—Susan III or oil Red oil O
—can be seen better with polarizer scope: Maltese cross
What is clinical significance of oval fat bodies?
—lipiduria = damage to the glomerulus caused by nephrotic syndrome
—tubular necrosis
—diabetes mellitus
—trauma case where fat from the bone marrow is released
Describe Bacteria found in urine
—not normal in urine
—urine is sterile, contaminated on way out
—wbc + bacteria = UTI
—nitrite test may be positive
—cocci and bacilli may be present
— refer to ranges and reports as 1+, 2+, 3+, 4+
What is clinical significance of bacteria in urine?
—presence of bacteria indicates a lower UTI (urethra or bladder) or upper UTI (ureter and kidney)
—most frequent organisms
——> Enterobacterales (GNR) (klebsiella, e. Cool, proteus sp., pseudomonas sp.)
——> Staphylococcus
——> enterococcus
Describe yeast in urine
—single, refractive, budding structures
—use reference range to report
— diabetic urine: high glucose and acid ideal for yeast growth
—immunocompromised and vaginal moniliasis
—acetic acid aids in identification of yeast cells (will lysis RBC)
Describe parasites found in urine
—most common: trichomonas vaginalis
——> pear shaped
——> “darts” around
— enterobius vermicularis = pinworm
—schitsisoma haematobium
Describe sperm found in urine
—urine toxic to Sperm so no movement
—may cause positive protein
—if seen in child, must be confirmed by supervisor
Describe mucous found in urine
—protein from glands ~ epithelial squamous cells of lower genitourinary and RTE
—Threadlike, low refractive index
—in females: no clinical significance
Describe casts found in urine
—unique to kidney
—formed in lumen of DCT and collecting duct
— take form of these lumens
—examine on low power to locate
— high power to identify
—report on lpf
— low light is needed
Describe cast composition
— major component: uromodulin (Tamm Horsefall) protein secreted from RTEs of the DCT and collecting duct
— rate of excretion is constant (can increase with stress and exercise)
—form during urine-flow stasis, acidity, high Na and Ca
What are the steps of cast formation?
PROTEIN MATRIX
1) aggregated uromodulin fibrils attached to RTEs
2) interweaving to form loose network, traps elements
3) more interweaving to form a solid matrix
4) attachment of elements to matrix
5) detachment of fibrils from RTEs
6) excretion of cast
Describe hyaline cast
—entirely uromodulin
— colorless
— low refractive index
—use low light
— normal range: 0-2/lpf
Describe clinical significance of hyaline casts
—non pathological
——> stress
——> exercise
——> fever and dehydration
——> heat exposure
—pathological
——> acute glomerulonephritis
——> pyelonephritis
——> chronic renal disease
——> congestive heart disease
Describe RBC casts
—bleed in nephron
—primary damage to glomerulus
—protein + dimorphic RBC
—orange-red in color
What is the clinical significance of RBC casts?
—may see granular, dirty, brown cast produced by hemoglobin breakdown
— see in acute tubular necrosis
Describe WBC casts
—most frequently composed of neutrophils
—look for granules and multilevel nucleuses
What is the clinical significance of WVC casts?
—appear infection and inflammation of the nephron
—-pyelonephritis (aid in identifying if lower or upper infection)
—acute interstitial nephritis (no bacteria)
—glomerulonephritis
Describe bacterial casts
—may be pure or mixed with WBC
— confirm with gram stain
— seen in pyelonephritis
Describe epithelial cell casts
— cast with RTE = advance tubular destruction
—urinary stasis with tubular lining disruption
—form in DCT
—round or oval
What is clinical significance of epithelial cell casts?
—see in toxicity if heavy metals, chemicals, drugs, viruses and transplant rejections
—pyelonephritis
—liver issues (hepatitis) bilirubin can stain the cast yellow
Describe fatty casts
— fatty cast+oval fat bodies+free floating fat= lipiduria= nephrotic syndrome
—seen in tubular necrosis, diabetes mellitus, crush injuries
—highly retractile
—
Describe granular cast
— fine or coarse (do not need to report)
—non-pathological: RTE lysosomes excrete in normal metabolism, after exercise and Activity
—pathological: disease states- granules could be a result of deterioration of cellular cast
— if stays in tubules too long, becomes brittle and turns into waxy cast
What are 2 results of extreme stasis?
Waxy cast
Broad cast
Describe waxy cast
— brittle, high retractile
—often fragmented with jagged ends and notches
—degenerated hyaline and granular cast
—extreme urine stasis
—renal failure
Describe broad cast
—renal failure casts
—destruction and widening of DCT
—formation in upper collecting duct
Describe crystal formation
—include: inorganic salts, organic compounds, and latrogenic (due to medication or treatment)
— change can occur due to: temperature, solute concentration, and pH
What are techniques used to identify crystals?
— most important —> pH
— size, shape, color
—normal or abnormal
—polarization
Describe amorphous urate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—Results from refrigeration and dissolved if warmed
— 5.5 pH
—normal
—will see pink sediment (uroerythrin) in tube
—looks like speckles
Describe Utica acid crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
— acidic
— rhomboid, whetstones, wedges, rosettes
—colorless to yellow-brown
—Highly birefringement
— seen in samples with high purines, high nucleic acids, leukemia patient receiving chemo, lesch- Nathan syndrome
Describe Na urates crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—normal but rare
—acidic
—colorless
Describe calcium oxalate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—most frequently seen
—normal
—dihydrate —> envelopes or X
—monohydrate—> oval or dumbbell
—both birefringement under polarized light
—foods: tomatoes, asparagus, and
ascorbic acid
—monohydrate crystals = antifreeze poisoning
Describe amorphous phosphate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—alkaline/neutral
—white precipitant that does not disappear after warming
—normal
Describe calcium phosphate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—normal
—alkaline/neutral
—not common
—colorless flat rectangular plates
—common constituent of renal stones
Describe triple phosphate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
— ammonium magnesium phosphate
—alkaline
—normal
— prism shape or coffin lids
—retractile
— no clinical significance
— seen with bacteria that split urea
Describe ammonium biurate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—alkaline
—normal
—yellow brown in acidic urine
—thorny apples
—indication of old sample
—associated with urea splitting bacteria
Describe calcium carbonate crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—alkaline
—normal
—dumbbell or spherical
—not significant
Describe cystine crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—abnormal
—acidic
— inherited metabolic disorder that prevents the reabsorption of cystine
—results in cystinuria. Forms stones at early age
—colorless hexagonal plates
Describe cholesterol crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—abnormal
—acidic
— rectangular plate with a notch in one or more corners
—highly birefringment
—see with lipiduria
Describe leucine crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—abnormal
—acidic
— yellow brown spheres with circles and striations
— seen less than tyrosine
— look like tree trunk life rings
—liver disease
Describe tyrosine crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
—abnormal
—acidic
—colorless to yellow
—needles
— liver disease
Describe bilirubin crystal including pH, shape, abnormal or normal, and If there is a condition related to crystal appearance.
— abnormal
— acidic
— clumped needles
—yellow
—liver disease
—viral hepatitis
Explain radiographic dye
— acid
—appears like cholesterol
— will result in very high specific gravity
Explain sulfonamide
—An antibiotic to treat UTI
—inadequate hydration will cause crystallization causing turbulence may damage
