LAB- Microscopic Examination Flashcards
Brightfield microscope:
Contrast is obtained by…
lowering the condenser and
stopping down the iris diaphragm
Requires a special condenser and objective with phase rings – produces a halo effect and increases contrast
Phase-contrast microscope
-Used for examining lipid material
-Lipids and cholesterol have a Maltese-cross appearance
Polarized microscope
Provides a three-dimensional image showing very fine structural detail
Interference-contrast microscope
Stain that provides clear delineation of structure and contrasting colors of the nucleus and cytoplasm
Sternheimer-Malbin stain (crystal violet and safranin O)
Stains triglycerides and neutral fats orange-red
Lipid stains: Oil Red O and Sudan III
What is the Hansel stain? and what is it used for?
Methylene blue and eosin Y stain eosinophilic granules
Stains structures containing iron
Prussian blue stain
Casts and crystals are recorded as number per ____ power field
low (x100)
Cells are recorded as number per _____ power field
high (x400)
How many fields should be examined in each case?
10
Reaction of body tissues to injury?
-Capillaries dilate and plasma proteins “leak” into the
surrounding tissue spaces
-WBCs are mobilized to combat the injury and repair the damage
-Neutrophils, monocytes, lymphocytes, occ. Eosinophils, histocytes, and macrophages
Congested, dilated glomerular capillaries
release
red cells and protein into the urine
With bacterial infection in the kidney or bladder,
neutrophils are…
mobilized and excreted w/
bacteria in the urine
___________ appear when a transplanted kidney is rejected.
lymphocytes
What can cause increased numbers of epithelial cells from the tubules?
-infections
-diminished blood supply
-tubules being affected by drugs or chemicals
What distinguishes kidney disease from lower urinary tract disease?
urinary casts
urinary casts only form in the kidney _______ and _________.
tubules, collecting ducts
contents of normal urinary sediment includes:
RBCs (_____ per hpf)
WBCs (_____ per hpf) – mostly neutrophils
Epithelial cells
Squamous and transitional cells
Casts (Hyaline)
Mucus
Crystals
Artifacts
Spermatozoa
0-3 (7 micrometers)
0-8
Increased ______ in urine may be one of the first
clinical signs of malignancy of kidney or bladder
RBCs
*presence is clinically significant!
When RBCs are seen with RBC casts, the source of hematuria is _______.
renal
(Inflammatory diseases, acute glomerulonephritis,
pyelonephritis, hypertension, bleeding due to trauma,
stones or tumors, and use of anticoagulants)
Red blood cells act as ___________ because they are altered by the specific gravity.
osmometers
How can RBCs and yeast be differentiated?
Yeast are oval in shape and show budding
Can use Eosin dye (RBC – red, yeast- colorless)
Can use acetic acid (RBC – lyse, yeast – no change)
WBC are best preserved in what type of urine?
acidic
What does greater than 50 WBCs suggest?
acute inflammation of genitourinary tract – culture should be done
> 50 WBCs accompanied with
proteinuria suggests….
pyelonephritis
> 50 WBC accompanied WITHOUT
proteinuria suggests…
lower urinary tract
infection (cystitis, urethritis, and prostatitis)
Neutrophils are most common WBC seen - multi lobed nucleus (_____ µm)
10-14
What can be confused with renal tubular cells?
Neutrophils.
acetic acid clarifies nuclei
Degenerated neutrophils are called “glitter cells” and is seen with…
hypotonic urine, and pyelonephritis
What may be the first sign of kidney transplant rejection?
Lymphocytes (6 to 9 µm)
What do monocytes suggest?
tissue damage
accompanied by severe inflammation (20 to 40
µm)
specialized white cells
formed in the reticuloendothelial system
-May contain inclusion bodies such as bacteria or
epithelial cells
-Suggest inflammation, immune mechanisms, or
other host defense reactions
Phagocytic histocytes
-Squamous (pavement) epithelial cells (40 to 60 µm w/ N:C ratio of ____)
1:6
Where do squamous epithelial cells originate?
terminal portion of the urethra in men and women, and the vagina in women
How do squamous epithelial cells appear when stained?
pink or violet
Occurrence of squamous epithelial cells is __________, and may indicate vaginal
contamination due to improper specimen collection
common
How are squamous epithelial cells reported?
Report as rare, moderate, or many per lpf (only cell to
report on lpf)
What is the second major epithelial cells?
Transitional epithelial cells (20 to 40 µm w/ N:C ratio of
1:3)
Where are transitional cells located?
Line the renal pelvis and calices, ureters, bladder, and
two-thirds of the urethra
Polyhedral in shape, appear pear-shaped, or oval with one or more nuclei
transitional epithelial cells
transitional cells are the same size as renal tubular cells, but may become larger
due to _________.
swelling
How do transitional cells appear when stained?
cells show dk. Blue nuclei w/ pale blue cytoplasm
Transitional cells are found in normal urine, but in large numbers these cells indicate…
disease of the bladder or renal pelvis, or the use
of a catheter
Renal tubular epithelial cells (12 to 20 µm w/ N:C ratio of
____)
1:1
What is the most clinically significant epithelial cell?
Renal tubular
Where do renal tubular cells come from?
Originate from the epithelial lining of the nephron’s tubular portions
Polyhedral in shape w/ slightly granular cytoplasm
renal tubular epithelial cells
When stained, appear purple w/ orange-purple cytoplasm
renal tubular epithelial cells
What cell type can undergo fatty degeneration – oval fat bodies?
renal tubular epithelial cells
What can >2 RTEs indicate?
acute tubular necrosis,
glomerulonephritis, acute infection, renal toxicity, and
renal infection
-Lipid-containing RTE cells
-Usually seen in conjunction with free-floating fat
droplets
Oval Fat Bodies
How is identification of oval fat bodies confirmed?
w/ Sudan III or Oil Red
O stains (orange-red), and the use of polarized
microscopy (Maltese cross formation)
What are oval fat bodies associated with?
nephrotic syndrome, acute tubular necrosis, diabetes mellitus, trauma cases where
bone marrow is released from the long bones
What does the presence of cytomegalic inclusion bodies suggest?
cytomegalic inclusion
disease, Hodgkin’s disease, lymphosarcoma,
leukemia, and aplastic anemia
What is it called when renal tubular cells have viral
inclusions in the nucleus of these cells?
Cytomegalic inclusion bodies
What is referred to as “owl-eye” cells?
When Cytomegalic inclusion bodies are stained w/ eosin or Giemsa stain, these
inclusion bodies appear separated by a clear halo
How do tumor cells appear?
Appearance varies, but most are identified on the basis of distinct nuclear changes
-Nucleus is much larger than normal
Tumor cells readily accept ___________ stain.
Papanicolaou
Tumor cells are usually accompanied by…
hematuria
> _______ bacteria per ml indicates a urinary tract infection.
Counts between _______ and _________ need to be repeated.
100,000
10,000 and 100,000
A count of _________ bacteria usually signifies urine contaminated from the
urethra or perineum
<10,000
Very low counts occur in patients on antibiotic therapy
Large quantities of bacteria accompanied by WBCs
and/or white cell or mixed casts indicate the presence of…
acute pyelonephritis
yeast can be found in patients with…
-diabetes mellitus
-urinary tract infections
-also common contaminate from skin and hair yeast
Characteristics of yeast that differentiates it from RBCs?
-ovoid
-higher refractive index
-budding
-does not take on eosin stain
a blood fluke that may be seen in the urine
Schistosoma hematobium
anterior flagellae and undulating membrane parasite
Trichomonas
-found in patients w/
urethritis or in the contaminated urine of
women w/ trichomonas vaginitis
parasite of the anus that may be seen during microscopic evaluation
Enterobius vermicularis
Protein material
produces by glands
and epi cells of the
lower GI tract and
RTE cells
Mucus (report per low power field)
Mucus consists of ___________ protein.
Tamm-Horsfall
Where are casts formed?
-distal convoluted tubule
-collecting duct
What are the two ways casts can be formed?
From the precipitation and gelling of the mucoprotein
found in the tubular fluid. Acid pH and high salt
concentration promote this gelling and precipitation
process.
From the conglutination of cells in the mucoprotein of
the tubular fluid
Casts are reported as number per ____ power field.
low
Hyaline casts are formed from…
plasma proteins and/or TammHorsfall mucoprotein (a protein gel secreted by
the RTE cells lining the distal portion of the renal tubule
-basis upon which all casts are formed
What is a normal amount of hyaline casts?
0-2 hyaline casts/lpf
Increased numbers of hyaline casts are associated with….
acute glomerulonephritis, acute pyelonephritis,
congestive heart failure, and chronic renal failure
Hyaline casts have a low refractive index. How are they stained?
Pale pink color w/ Sternheimer-Malbin stain
Hyaline casts dissolve easily in what type of urine?
alkaline
Few appear in standing urine (>1 hr) since bacterial
contamination causes urea to decompose to form
ammonia, an alkaline medium
Red blood cell casts almost always indicates…
renal disease
How do RBC casts appear stained and unstained?
High refractive index
Unstained are yellow-orange in color
Stained are pale lavender
Red blood cell casts usually occurs in diseases where what has been damaged?
the basement
membrane of the glomerulus has been damaged, thus
allowing RBC passage into Bowman’s capsular space
What are two diseases RBCs are associated with?
acute glomerulonephritis and lupus nephritis
RBC casts are always observed with?
free-standing RBCs
What are WBC casts commonly called?
“pus” casts
WBC casts can be difficult to distinguish from __________ casts.
epithelial
WBC casts are always observed with…
free standing WBCs
How do WBC casts appear?
-highly refractive
-Degeneration may cause granular appearance
-Can be difficult to differentiate from Epithelial
casts
WBC casts can be reported as either cellular or _________.
cellular-granular
What color are WBC casts when stained?
purple to blue-green
What is the most common cause of WBC casts?
acute pyelonephritis***
Acute pyelonephritis is usually accompanied by…
large number of WBCs
in sediment, proteinuria and bacteriuria
WBC casts play a secondary role in….
acute
glomerulonephritis
Casts may contain a few fat droplets w/in a hyaline
matrix, or may be filled w/ fat globules of different sizes
Fatty Casts
-Fat is globular in shape, brownish-tan in color, and highly refractive
Fatty casts are best recognized using a __________ microscope, which gives
lipid globules a distinctive Maltese cross configuration and double refractive appearance
polarized
When fatty casts are accompanied by passive proteinuria, associated with…
nephrotic syndrome
-Also seen w/ diabetes mellitus, and poisoning from mercury or ethylene glycol
Consist of renal tubular epithelial cells
-Highly refractive, with single large round nucleus
Epithelial Cell Casts
How do Epithelial Cell Casts appear when stained?
have a purple nuclei surrounded by pale pink matrix
Epithelial Cell Casts are usually arranged in a symmetric manner,
indicating…
damage to an entire portion of the tubule
When in random manner, indicates damage to
different and separate portions of the tubule
Epithelial Cell Casts suggests…
intrinsic kidney disease involving the renal tubules
Glomerulonephritis, vascular disease, exposure
to a toxin, or presence of a virus
Injured cells may contain inclusions which suggest the primary injury affecting their
degeneration
Regarded as those casts which have at least two well defined and distinct portions
Ex: part epithelial and part granular, or part granular and part waxy
Mixed Casts
The presence of different cellular constituents in a cast often implies that…
more than one part of the nephron has
been injured
Ex: if both red and white blood cells appear in a
cast, both glomerular and interstitial damage
may be indicated
Opaque and are characterized by the
presence of course or fine granules
Some casts may represent degeneration of cellular casts
Shrinkage and fragmentation of cell nuclei,
membrane disruption, and granulated cytoplasm
Granules contain aggregates of plasma
proteins
Granular Casts
Granular casts accompany heavy….
proteinuria (nephrotic syndrome)
Granular casts could indicate…
orthostatic proteinuria,
congestive heart failure w/ proteinuria, as
well as chronic or acute renal disease
What is the order of different cast formation?
Cellular cast → coarsely granular cast → finely granular cast → waxy cast
When stained, course granules are _____ purple, and fine granules are a ______ purple color
deep, light
Waxy casts represent final phase of cellular degeneration
Identified easily due to sharp refractive outlines, and “broken off” ends
When stained, appear light to dark _______
purple
What are waxy casts associated with?
-extreme urine stasis, seen with chronic renal disease/failure, or kidney
disease resulting from diabetes mellitus
-Numerous waxy casts indicate a fairly long
renal transit time
Broad casts are Larger diameter and believed to be formed in __________ which have undergone dilation and
destruction
renal tubules
What is referred to as “renal failure casts”?
Broad casts
Broad casts indicate __________ renal diseases, caused by hypertrophy of tubules and ducts, which results in excessive urea absorption
Acute tubular necrosis, severe chronic renal disease,
and urinary tract obstruction
Prognosis is grave when found in numbers
degenerative
Are most urinary crystals significant?
appear in normal urine and most are NOT clinically significant (only a few are)
What are normal crystals that can be seen in acid urine?
Amorphous urate, uric acid, and calcium oxalate crystals
What are normal crystals that can be found in alkaline urine?
Amorphous phosphates, triple phosphate, ammonium biurate, calcium phosphate, and calcium carbonate crystals
Consists of calcium,
magnesium, or
potassium salts
Appear colorless or
yellow-brown
Amorphous urate
crystals
crystal that is readily soluble upon warming to 60°C and in alkaline substances
Frequently appear in
sediment and not
considered significant
Amorphous urate
crystals
Crystal that is Common in small numbers, but large numbers may indicate gout
Uric acid crystals
What are uric acid crystals soluble in?
in alkali, but
insoluble in alcohol and
acids
How do uric acid crystals appear?
– square, diamond-shaped, prism shaped, wedge-shaped, or grouped in rosettes
Colored yellow-brown or
red-brown
What are Calcium oxalate crystals soluble in?
dilute hydrochloric acid
Calcium oxalate crystals are mostly seen in…
acid urine (occasionally in neutral, or alkaline urine)
How do calcium oxalate crystals appear?
-Appear octahedral or
dumbbell-shaped
-Double refractive index
Calcium oxalate crystals in large numbers are associated with…
the formation of urinary calculi
Small, colorless
granules
Readily soluble in dilute
acetic acid
Usually nonpathologic
Amorphous phosphate
Triple phosphate (ammonium magnesium) crystals are seen mostly in what type of urine?
alkaline urine
(occ. In neutral, and acidic
urine
How do Triple phosphate (ammonium magnesium) crystals appear?
colorless, three-tosix-sided prisms (“coffin-lids”),
or as feathery crystals shaped like fern leaves
Triple phosphate (ammonium magnesium) crystals readily dissolve in…
dilute acetic acid
What are Triple phosphate (ammonium magnesium) crystals associated with?
Usually nonpathologic, but
may be associated w/ urine
stasis and chronic urinary tract infection
Appear as a yellow brown spicule resembling a thorny
apple, or a yellow brown sphere
Ammonium biurate crystals
What dissolves ammonium biurate crystals?
acetic above 60°C or in
strong alkaline solutions
What are ammonium biurate crystals associated with?
Presence does not
indicate disease
Appear as colorless
prism or rosettes
Calcium phosphate
crystals
What are Calcium phosphate crystals soluble in?
dilute acetic acid
What are calcium phosphate crystals associated with?
Usually not pathologic,
but may be associated
w/ urine stasis and
chronic infection of the
urinary tract
What type of urine are calcium carbonate crystals found in?
Usually found in alkaline
urine, but also seen in
neutral or faintly acidic
urine
Appear colorless and
amorphous (dumbbells,
rhombi, and needles)
Calcium carbonate crystals
What does calcium carbonate dissolve in?
acetic acid
What are calcium carbonate crystals associated with?
Usually nonpathologic,
but large numbers may
indicate inorganic calculi
formation
When tyrosine or leucine are found alone what does it indicate?
When found together?
-rare inherited metabolic disturbance
-usually signify severe liver
disease
highly refractile – appear as thin ,dark needles arranged in sheaves or clumps – may be colorless, but usually yellow due to bilirubin
Tyrosine crystals
What are tyrosine crystals soluble in?
alkali, mineral acid, and acetic acid
highly refractile – appear as
yellow to yellow-brown spheres w/ radial striations
Leucine crystals
What are leucine crystals soluble in?
hot alcohol and alkali
Cystine crystals are rarely seen except with…
an inborn error of metabolism called cystinuria
Renal tubules fail to reabsorb cystine and three other amino acids – cystine crystallizes and forms renal
calculi
How do cystine crystals appear?
Highly refractive clear hexagonal plates
What are cystine crystals readily destroyed by?
bacteria
What are cystine crystals soluble in?
Readily soluble in alkali (ammonia and dilute
HCL)
Appear as star
-shaped clusters of needles,
rhombic plates, or elongated prisms
–may be colorless, or yellow/brown
Hippuric acid crystals
What are Hippuric acid crystals soluble in?
hot water, alkali, and ether
What are Hippuric acid crystals associated with?
Generally nonpathologic and have little diagnostic
significance
What do bilirubin crystals indicate?
hepatic disease
Appear red-brown,
shaped like needles or
rhombic plates
Bilirubin crystals
Bilirubin crystals may alter the color and structure of other crystals, particularly _________ crystals.
uric acid
What are bilirubin crystals soluble in?
acids, alkali and
chloroform
Cholesterol crystals usually appear with…what disease
chyluria, or w/ severe urinary
tract infections
Appear as colorless, transparent plates w/ regular or irregular corner notches
Cholesterol crystals
What can cholesterol crystals be easily detected by?
e Lieberman-Burchard
reaction
addition of acetic anhydride and concentrated sulfuric
acid to an ether extract of urine – green color
What are cholesterol crystals soluble in?
Highly soluble in chloroform and ether
Appear as colorless
prismatic or star-like
forms
Confirm w/ 1 ml of 10%
ferric chloride – purple
Urine should be heated
gently to remove any
acetone bodies, in
order to prevent false
positive results
Aspirin crystals
Seen in the urine of
children, and in adults
suffering from
progressive destruction
of muscle tissue due to
muscular dystrophies,
atrophies, and myositis
Appear as
pseudohexagonal plates
Creatine crystals
Caused by
crystallization of
sulfonamide drugs
Variety of forms, seen
as dumbbells,
asymmetrical sheaves,
rosettes, or hexagonal
plates – colorless or
yellow-brown
Sulfonamide crystals
What are Sulfonamide crystals soluble in?
acetone
What do ampicillin crystals look like?
-Crystallized penicillin in
acid urine
Seen w/ large doses of
penicillin
Colorless long and thin
crystals
Crystallized meglumine
diatrizoate in acid urine
X-ray crystals
-Appear as long, thin,
clear rectangles; or as
flat, four-sided notched
plates
Soluble in 10% NaOH
solution
Causes very high
specific gravities
X-ray crystals
Normal pH of urine is
4.5-7.8