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ACTIVITY 7: MICROSCOPIC ANALYSIS OF URINE
MICROSCOPIC ANALYSIS
● Third partof urinalysis
● For detection ofinsolublematerialsinurinesuch
ascells, casts, and crystals
● Abnormalities in the physical and chemical
portionsoftheurinalysisplayaprimaryroleinthe
decision to performamicroscopicanalysis,thus
theuseoftheterm"macroscopicscreening" for
cost- effectivity. (applicable only in some
laboratories depending on their SOPs)
First Part - Physical Examination
Second Part - Chemical Examination (Routine Reagent
Strip)
ThirdPart-MicroscopicAnalysis-useforthedetectionof
insoluble materials in your urine and what are those F irstforthescreeningtest,wecanseeadifferenceinthe
insoluble materials? - these are your cells, cast, and color of your urine.
crystals. - Areddishcolorofyoururinemayhaveaclinical
significance and that significance or correlation
icroscopic analysis serves as your confirmatory testor
M may be the presence of blood in your urine
thecorrelationtesttodeterminewhetherornottheresults sample.
of the physical or chemical examination are trueornot
remember when you will be releasing result in your bnormal clarity or turbidity in the urine may be
A
physical and chemical analysis of your urine in the first correlated with the presence of hematuria,
part of your examination process in your routine urinalysis hemoglobinuria,and,myoglobinuriaandinorderforusto
determined what is the cause or thepathologicornon
take note: that a result of your microscopic analysis pathologiccauseofyourchangesintheurineturbidityor
beingthethirdpartofurinalysismustcorrelatewiththose clarity we must performed your microscopic analysis.
previous procedures .
T hepresenceofbloodassignifiedtheapositivebloodin
❖ R ememberyouwillperformmicroscopicanalysis the urine reagent strip may have a clinical significance
based on your theoretical aspect of this and correlation withthemicroscopicanalysis,whenyou
particular discussion. will see a four positive blood, remember in your urine
❖
Whenever there is a flag or abnormality in the reagent strip you might see a presence ofRBCorsome
resultportionofyourphysicalandchemicalparts RBC cast in your urine.
of your routine urinalysis this is done in order to
have acauseeffectivitybutitisonlyapplicable presence or the positive protein test in your urine
A
only in some laboratory depending on their reagent strip, you may see or you may correlateitwith
standard operating procedures. seeing a cast as well as cell in your urine.
❖
ForthelaboratorieshereinthePhilippinesweare
performing microscopic analysis whether or not T he presence or the positive result inyoururinereagent
theresultofthephysicalandchemicalportionof stripfornitritemaysignifythat youcanseeabacteriaor
your urinalysis turned as the macroscopic WBCs in the microscopic analysis of your urine.
screening are normal or abnormal, whether or
not the result is normal or abnormal we will be nother one leukocyteesterasewhichispositiveinyour
A
performing microscopic analysis in the urine reagent strip you may see using your microscope
laboratories here in the philippines. some WBCs, WBC casts, and bacteria.
nother correlation between your microscopic and
A
macroscopic screening may be in the form of your
positiveurineglucose.Wheneverthereisapositiveurine
BMLS 3-1 1
lucose expect that you will see some yeast in your
g ● F or detection of casts use low powerfeltoryou
microscopic analysis. lowpowerobjective(especiallyontheedgesof
theglassslide),foridentificationofcastsusehigh
URINE SEDIMENT PREPARATION power objective or your high power field
- You might see some casts in the edges of the
● T ransfer 10-15 mL of urine in a test tube glass slide because they have a high specific
(average/recommended volume12mL gravity or mass so therefore when you will be
● centrifuge at400RCFfor5minutes-why?-itis coveringyourslidewiththeurineorwiththeurine
use to prevent this integration of the form sediment with your coverslip some casts might
elements in the urine. travel from the center of the urine towards the
- Over Centrifugation remember may cause the edges of the glass slide.
disintegrationoftheformedelementsintheurine,
because the are very sensitive to high amounts ADDIS COUNT
of centrifugal force. - uses light or bright field microscopy it is used
● Decanturine(0.5or1.0mLmustremaininthetest usingyourlight microscopyforyoutobeableto
tube) - do not be conscious enoughto decant determine the addis count of the patient.
your urine because remembered when you will ● Quantitativemeasurementofformedelementsof
decant your urine whether it is fastorslowyou urine usinghemocytometer
will be having a remaining portion ofyoururine ● Specimen:12-hr urine
whichyoucanresuspendyoursedimentsdonot ● Preservative:formalin
be conscious of that. Normal Values
● Transfer20_uLor0.02mLsedimenttoglassslide RBCs:0-500,000/ 12-hr urine
with22mmx22mmcoverslip-itisveryimportant WBCs and Epithelial Cells:0-1,800,000/ 12-hr urine
for you to have a coverslip in order for you to Hyaline Casts:0-5,000/ 12-hr urine
prevent the urinefromstickingfromthedifferent
objectives of your microscope. CORRELATIONS
● Examine microscopically (1 0 LPF,10 HPF under
reduced light) - why?-rememberyoumustuse
areducedorsubduedlightformicroscopywhy?-
becausewhen youwilluseasubduedlightyou
may see some different sediments or form
elementsinyoururinethathaveahighrefractive
index and this are your cast and crystals. Your
castandcrystalshavehighrefractiveindexsoif
you will use a full light a full blast light in the
microscopic analysis you might be abletomiss
them.
URINE SEDIMENT PREPARATION
● If <12 mL of urine is available for microscopy,
centrifuge3mL of it
● If<3mLofurineisavailable,examinesediment
without centrifugation why? - centrifugationand
removing the supernatant might leave no
sediment suspension and centrifuging low If you will be seeing RBCs in your microscopy you will
volumes of urine might cause for there expect a turbidity as well as a red color of urine
disintegration of the formed elements in the urine. signifyingthepresenceofRBCs,forthechemicalportion
● Conversion of RPM to RCF: RCF = 1.118 x 10-5 x youwillbeabletoseeapositiveresultinthebloodand
radius in centimeters × RPM2 protein reagent strip.
- Thisisfortheconversionofyour400RCF
toRPM.Typicallyweareperforming1800 BC’s, if you are to beseeingWBCsinyoururineunder
W
RPM to 2000 RPM as the centrifugation themicroscopeyouwillexpectthatthereisaturbidityin
speed for your microscopic analysis. your urine and there is a positive protein nitrite and
leukocytes esterase and those signifies that the patient
might have a UTI.
BMLS 3-1 2
SEDIMENT EXAMINATION TECHNIQUES: SEDIMENT STAINS
F orepithelialcellsspeciallyforwomenthatarehavinga
menstrual cycle. There wouldbeanincreasedepithelial
cell and you will be seeing a turbidity in your physical
exam of your urine.
F or the cast ontheotherhandyouwillbeabletoseea
positive protein because your casts remember are
typically made up of protein.
F or bacteria you will be seeinganincreasedturbidityin
e haveotherurinesedimentstainsthatareusedinthe
W
the physical part of your routine urinalysis. While for the
automatedmicroscopicmachinesthatarepresentinthe
chemicalreagentstripforyoururinewheneverthereisa
urinalysis section or the clinical microscopy section of
past presence of bacteria you will be able to see an
your clinical laboratory and thisisyourphenanthridine-
increasepHortheurinemayhaveaalkalinepHsomeof
has an orange color and this stains your DNA. And we
your bacteria are capable of reducing your urea to
also have your carbocyanine which has a green color
ammoniathroughtheenzymeureaseandammoniatake
and it is use to stain nuclear membrane mitochondria
noteisaveryalkalinesubstance.Andwheneveryouwill
and cell membrane and this are bothuseinthesismics
see bacteriainthemicroscopyyoumightexpecttosee
uf-100 urine cell analyzer this two stain are used in the
a positive result for your nitrite and leukocytes and that
automated machines use for microscopy.
may signify that thereisapresenceofyoururinarytract
infection in the patient.
MICROSCOPIC TECHNIQUES
F or crystal, you may have an increase in turbidity you
may correlate it with an increase in turbidity of your
physical exam of your urine and in the identification of
crystals take note the pH portion of your chemical
reagentstriporthechemicalanalysisofurinemighthave
a significant effect or a significant health in the
identification of your crystals why? - because some
crystals may formed and acidic pH and some crystals
might formed in your alkaline pH and also you can
correlatetheidentificationofyourcrystalsinthecolorof
yoururineaswellasthedifferentresultsofyourchemical
reagent strip. For example: if the patient hasapositive
bilirubin in the chemical part of your routine urinalysis
expect that you will see bilirubin crystals in the
e have the most common microscope to use in the
W
microscopy, part of your routine urinalysis.
routineurinalysisspecificallyformicroscopicanalysisand
this is yourbright-field microscopy.
EMEMBER!hemoglobininRBCswhichisequivalentto10
R
milligrams for deciliter may cause a positive reactionin
emember for polarizing microscopy it is use toidentify
R
your routine strip.
cholesterol in oval fat bodies , fatty casts, and crystals
somecrystals mightpresentbyfringesandsomeformed
elements such as your oval fat bodies might present
birefringenceandthisbirefringence maybeseenandthe
succeeding parts of your powerpoint presentation.
ther additional note for the interference-contrast
O
microscopy we are using two types ofyourmicroscope
we have your nomarski microscopy technique which is
the differential interference-contrast and we also have
your hofmann microscopic technique which is your
modulation-contrast microscopy.
BMLS 3-1 3
MICROSCOPIC TECHNIQUES: POLARIZING MICROSCOPY ED BLOOD CELLS
R
Sources of error:
● Yeasts
● Oil droplets
● Air bubbles
● Calcium oxalate monohydrate crystals
Remedy:
● Add2% to lyse the RBCs
Clinical Significance
T his is an example of the birefringence shown in the Glomerular damage, vascular injury inGUtract,
microscopic image. The first part is your ovalfatbodies malignancy, renal calculi, trauma
whichpresentbirefringenceintheformofamaltesecross - We have different sources of error for your red
pattern. blood cells. You might be confused with the
identification of your red blood cells with your
nd the other one is your uricacid,bothofthisformed
A yeasts, oildroplets,airbubbles,calciumoxalate
elementsintheurinemightbyrefringenceandtakenote monohydrate crystals. How are you able to
by refringence is typically seen when you will be using remediate that particular source of errororhow
yours polarizing microscopy as a microscopic technique. areyoubeingaidedwiththedifferenttechniques
foryoutobeabletoconfirmwhetherornotthat
SEDIMENT CONSTITUENTS isanRBC?-foryoutoremedy thaterroryoumay
add2% aceticacidtolyseyourredbloodcells.
RED BLOOD CELLS(also known as erythrocytes) When you are lysing your redbloodcells take
● NV: 0-2/0-3/HPF - remember for the detection note using your 2% acetic acid all of this
andthereportingofyourredbloodscellsyouwill sediment constituents such as your yeast, oil
useyourhighhighpowerfield(HPF)theaverage droplets, air bubbles, and calcium oxalate
number per HPF 0-2/0-3 that is the normal value monohydrate crystals may “dko maawatan”
● Most difficult to be recognized by newbies 22:47.Remember2%aceticacid can onlylyse
● Smooth, non-nucleated, biconcave disks yourRBCs butnotothersourcesoferrorforyour
● Hypertonic:-expectthatyouwillseeacrenated RBCs. And the otheronepointof thedifference
RBC. between your yeasts in your RBCs is that your
● Hypotonic: - you will be expected to see a yeastsyoumayseedifferent buddingpatternsor
swollen or enlarged RBC. your budding forms and this may aid in the
● Glomerular membrane damage: dysmorphic, distinguishingofyouryeastswithyourRBCs.when
with projections, fragmented wesaybuddingyoumayseeyeastswithanother
yeastscellinthebuddingformattachedtoitand
thisisseenintheimagepresentedandthisisthe
budding yeasts.
BMLS 3-1 4
HITE BLOOD CELLS (also known asleukocytes/granular
W HITE BLOOD CELLS
W
cells) Eosinophils-takenotesotheuseofyourhanselstainyou
● NV:0-5/0-8/HPF-rememberyouwillseedifferent may be able to identify your eosinophils because your
white blood cells in the urine present with a hansel stain will stain your eosinophilic granules . the
granule another point of distinguishment eosinophilsI'mtalkingaboutareseeninthemicroscopic
between the white blood cells and other forms image.
elements is the presents of the granules. ● NV: <1% of WBCs
● Increased number: infection/inflammation ● >1%: associated with interstitial nephritis
HITE BLOOD CELLS
W HITE BLOOD CELLS
W
Neutrophils Mononuclear Cells (lymphocytes, monocytes,
● Most predominant macrophages, and histiocytes)
● Granulated and multilobed ● Normally present in small numbers
● In hypotonic urine, they swell and granules ● Increased lymphocytes: renal transplant
undergo brownian movement producing a rejection
sparkling appearance, AKA glitter cells - no ● Increased monocytes and histiocytes: chronic
pathologic correlation inflammation and radiation therapy
● ·When dying, they form blebs and fingerlike
projections and this is now turn as your ghost PITHELIAL CELLS
E
cells Squamous Epithelial Cells
● Clinical Significance:Bacterial infection ● Largest cell with abundant irregular cytoplasm
and prominent nucleus
- Inorderforustodifferentiateyourglittercellswith ● Cell size is about 30-50um (5-7x size of RBCs)
another leukocytes with your other leukocytes ● Nucleus is about the size of RBC
your normal leukocytes your glitter cells when ● Fromliningsofvagina,femaleurethra,andlower
stainwithsternheimermalbinstainitwillbestain male urethra
aspaleblue whilefortheotherleukocyteswhen VARIATIONS:
your other leukocytes stain with sternheimer ● Cluecells- covered withgardnerella vaginalis
malbin stainit will stain pale pink ● Associated with bacterial vaginosis
Squamous Epithelial Cells - seen in the microscopic
image is the normal squamous epithelial cells theother
one is the variationofsquamousepithelialcellswhichis
yourcluecellnoticethatthefamousepithelialcellwhich
isturnasyourcluecellislinewithyourdifferentbacteria
known as yourgardnerellavaginalisanditisassociated
with bacterial vaginosis
BMLS 3-1 5
T ransitional Epithelial Cells (also known as enal Tubular Epithelial Cell (R.T.E cell) - this are your
R
europhilial/bladder cells) renal tubular epithelial cells observed under the
● About 20-30um (4-6x the size of RBCs) microscopenoticetheeccentricallyplacenuclateorthe
● Spherical, polyhedral or caudate with centrally location of your nucleus which iseccentricornotinthe
located nucleus center
● From renal pelvis, calyces, ureter, urinary
bladder, and upper male urethra (those parts
capableofexpandingtoholdurine)-thatiswhy
your transitional epithelial cells are capable of
changingitsmorphologyinorderto caterforthe
expansionorthechangesintheshapeorsizeof
this different parts of your urinary tract.
● Increased following catheterization (singly, in
pairs, or clumps/syncytia)
● If exhibiting abnormal morphology- malignancy
or viral infection
ARIATIONS
V
Oval Fat Bodies
● Lipid containing RTE cell (may also be a
monocyte or macrophage)
● Seen in lipiduria (e.g. disease state which may
contribute to a urine having lipids is your
nephrotic syndrome)
● Identified by: Lipid stains; and Polarizing
microscopy(maltese crossformation)
T his is your oval fatbodieswhichhasyourbirefringence
or presenting an maltese cross appearance under your
polarizing microscope.
Renal Tubular Epithelial Cell (R.T.E cell)
● Most clinically significant E.C.
● Origin:nefrol
● 3-5x the size of RBCs
● Rectangular, polyhedral, cuboidal, or columnar
with eccentric nucleus
● ·Sometimes bilirubin-stained or
hemosiderin-laden
● RTE cells from DCT may be mistaken forWBC
● >2 RTE/HPF indicates:tubular injury
Bubble Cells
● RTE cell with non-lipid vacuoles
● Injured cellsinwhichtheendoplasmicreticulum
has dilated prior to death
● Seen inacute tubular necrosis
BMLS 3-1 6
● andida albicans seen in DM and vaginal
C
T his is your bubble cells notice the non lipid vacuoles moniliasis
present in the cytoplasm of the bubble cell forming
bubblesthisarenonlipid vacuoles.Essentiallythisisyour
endoplasmicreticulumthathasdilatedpriortodeathand
it is very significant of patient having acute tubular
necrosis.
ARASITES
P
BACTERIA Trichomonas vaginalis
● True UTI: whenever there is a true neurani tract ● Most frequently encountered
infection you expect to see bacteria as well as ● Pear-shaped flagellate with jerky motility
your WBCs in your urine. ● Agent of pingpong disease/strawberry cervix
● Specimen contamination/old specimen:youwill ● Reported as rare, few, moderate, or many/HPF
only see bacteria in your urine ● When not moving, may resemble WBC,T.E.C,or
● Most common cause of UTI: is your escherichia RTE cell
coli
● Presence of motility for your bacteria your
bacteria being living organisms are capable of
beingmotiledifferentiatesthemfromamorphous
materials (crystals)
Enterobius vermicularis egg
● Most common fecal contaminant - because
remember when there is a presence of
enterobiusvermicularisegg inyoururinethisisa
hallmark that your urine is contaminated with
disease.
YEAST
● T rue yeast infection: combination of yeasts as
well as WBCs in your urine.
● Small, refractile oval structure that may or may
not bud - remember budding yeasts may be
points of differencebetweenyouryeastsaswell
as your RBC
● Branched mycelial forms seen in severe
infections
BMLS 3-1 7
Schistosoma haematobium egg ● ifference from cast: irregular shape
D or
● Blood fluke witha terminalspine appearance
● Causes hematuria ● No clinical significance
● Associated withyour bladder cancer
CASTS
● E xcretion is termedcylindrorhea
● Unique to the kidney -becausetheurinarycast
are formed in nephrons
SPERMATOZOA ● Represents a biopsy of an individual tubule
- yourspermatozoaistypicallyseenintheurineof ● The most difficult and most important urinary
your male patients but sometimes in female sediment constituent
patients you will see aspermatozoaintheurine ● Formed in the: distal convoluted tubule
of your female patients especially after that ● Major constituent:: Tam-horsfall protein or
femalepatientishavingyoursexualintercourse neuromodulin (produced by renal tubular
andthepresenceofspermatozoaintheurineof epithelial cells.)
a female patient may be significant and the ● Other constituents: albumin, and
determination of your medicolegal cases immunoglobulins
especially for rape. ● Factors affecting formation: urine stasis, acidity,
● Oval with slightly tapered head presence of sodium and calcium
● Long, flagella-like tail ● Casts should have an even anddefiniteoutline,
● After ejaculation/ sexual intercourse parallel sides, and two rounded ends
- Thereportingdepends onthelaboratory ● Hasuniformdiameter(about7-8xthediameterof
SOP but it itis acaseofamedicolegal RBCs)
especiallyforrapecasesyoumustreport ● Examination is performed along the coverslip
this cell or sediment constituents in the edges with subdued light
urine. - you may miss a cast when using a full
blast light microscopy.
● Cylindroids-castwithtail,formedinloopofhenle
same significance as casts
CAST
MUCUS THREAD
● Has low refractive index
● Major constituent: Tam-Horsfall protein also
known as neuromodulin
BMLS 3-1 8
BLOOD CAST/ MUDDY BROWN CAST
● Contains Hgb from lysed RBCs
CASTS: CAST FORMATION ● Homogenous appearance with orange red color
1. Aggregationofuromodulinproteinintoindividual ● Same significance with RBC cast
protein fibrils attached to the RTE cells
2. Interweaving of protein fibrils to form a loose
fibrillar network (urinary constituents may
become enmeshed in the network at this time)
3. Further protein fibril interweaving to form asolid
structure
4. Possibleattachmentofurinaryconstituentstothe
solid matrix
5. Detachment of protein fibrils from the epithelial
cells
6. Excretion of the cast
WBC CAST
HYALINE CAST
-s eeninpatientswhichhaveundergonestrenuous
exercises. Your hyaline cast is consideredtobe
theprototypecastandthebeginningofalltypes
of your cast
BC CAST
R
PSEUDO LEUKOCYTE CAST
● Not a true cast (do not report as cast)
● Clumps of WBCs
● No cast matrix
● Seen in lower UTI - WHY?
- Remember that casts are formed in the
upper parts of your urinary tract. Renal
tubularcellssecreteTAM,Horsfallprotein
or uromodulin.Therefore,whentheUTIis
onthelowerurinarytractprimarilyinthe
bladder termed as cystitis, no TAM,
Horsfall protein or uromodulin will be
released
BMLS 3-1 9
GRANULAR CAST
BACTERIAL CAST
● Identified using Gram stain
two types granular cast coarsely granular cast finely
granular cast: are characterized as a cast having your
sandpaper appearance.
FATTY CAST
EPITHELIAL CELL CAST
BMLS 3-1 10
WAXY CAST Factors that contribute in crystal formation
● pH
● Solute concentration
● Temperature
Theprimaryreasonfortheidentificationofurinary
crystalsistodetectthepresenceoftherelatively
few abnormal types that may represent such
disorders as liver disease, inborn errors of
metabolism, or renal damage caused by
crystallization of compounds Within the tubules.
Take note:
BROAD CAST ● Solutes precipitate atrefrigeratedtemperature
● Often referred to asRenal failure cast ● In the identification of crystals, the specimens
● Indicates destruction/widening of tubular walls must be atroomtemperature
● Any types of cast canbebroad(mostcommon
are granular and waxy) NORMAL ACID CRYSTALS
● 2-6x wider than ordinary cast AMORPHOUS URATES
● Significance: Extreme urine stasis, renal failure ● Fluffy orange or pink sediment (also known as
your brick dust) due touroerythrin
● Yellow-brown granules (microscopic)
● Resembles granular casts (pseudocasts)
● Turns into uric acid after addingacetic acid
● Turns into ammonium biurate after adding
ammonium hydroxide
● Increased in gout and chemotherapy
● Soluble in heat and alkali
OTHER CASTS Reading Assignment
● Pigmented Cast
● Mixed Cellular Cast
● Crystal Cast
CHEMICAL OR UNORGANIZED SEDIMENT
HEMICAL OR UNORGANIZED SEDIMENT
C
CRYSTALS
● Excretion in urine is termed as:crystalluria. URIC ACID
● Mostrecognizedbutthemostinsignificantpartof ● Product of purine metabolism
urine sediment ● Most pleomorphic crystals -becauseitassumes
● Formed by precipitation of: salts, organic a wide variety of shapes, and these are the
compounds, and medications different shapes that your uric acid,
● Crystals in fresh urine is most frequently ● Shapes: Rhombic/diamond (most common), 4
associated with specimens with in
creased sided flat plate (whetstone), lemon-shaped,
specific gravity wedges, barrel, rosettes, irregular plates,
● Crystals are usually reported as rare, few, laminated forms.
moderate, or many perhigh power field ● Hexagonal forms are mistaken ascystine crystals
● Abnormal crystals may be averaged and ● Increased in Lesch-Nyhan Syndrome, gout, and
reportedlow power field chemotherapy
● Soluble in alkali
BMLS 3-1 11
CALCIUM SULFATE
● Cigarette butt appearance
● Soluble in acetic acid
OTHER CRYSTALS (READING ASSIGNMENT)
● Hippuric Acid
● Acid Urates
● Monosodium Urates
CALCIUM OXALATE
● Most frequently seenin normal acid crystal
● Dihydrate (also known as WEDDELLITE)
○ assumes an envelope octahedron,
bipyramidal.
○ consideredtobethemostcommonform
of your calcium oxalate crystal.
○ Your dihydrate is also known as your
wedelite, and it assumes an envelope
octahedral bipyramidal, and it is
•considered to be the most common
shape or most common form of your
calcium oxalate crystal.
● Monohydrate(alsoknownasWHEWELLITE)):Oval,
dumbbell
ORMAL ALKALINE CRYSTALS
N
● Increased in foods rich in ascorbic acid
AMORPHOUS PHOSPHATES
- Itisincreasedinfoodsrichinoxalicacid
● Most common cause of turbidity in alkaline urine
such as tomato, asparagus, and
● Fine, lacy-white precipitate (macroscopic)
ascorbicacid,anditisalsoincreasedin
● Granular (microscopic)
ethylene glycol poisoning or
● Soluble in dilute acetic acid
methoxyurethane poisoning.
● HOW TO DIFFERENTIATE WITH AMORPHOUS
● Increased in ethylene glycol poisoning or
URATES?
methoxyurethane poisoning
- Through the use of your pH, and the
● Soluble in dilute HCI
sediment color, and your solubility.
Remember that amorphous phosphates
are soluble in dilute acetic acid, while
your amorphous urates are soluble in
alkaline.
BMLS 3-1 12
AMMONIUM BIURATE
● Alkalinecounterpartofuricacidandamorphous MAGNESIUM PHOSPHATE
phosphates ● Colorless, elongated, rectangular, rhomboid
● Yellow-brown thorny apple shape crystal. plates
● Seen inall specimens, ● End or corners may be notched
● Increased in the presence of urea- splitting ● Edges are irregular or eroded
bacteria ● Soluble in acetic acid
● Turns touricacidafteraddinghydrochloricacid
or acetic acid,
● Soluble in acetic acid with heat
CALCIUM PHOSPHATE
● Dibasic calcium phosphate (a lsoknownasyour
stellar phosphate) appears colorless, flat plates,
thinprisms,inrosetteformswhichmayresemble
ORMAL ALKALINE CRYSTALS
N sulfonamide crystal
TRIPLE PHOSPHATE ● Monobasic calcium phosphate appears
● AKAammoniummagnesiumphosphate,strovite, irregular, granular sheets or plates, less common
and your municipal hall crysta ● Other forms: Hydroxyapatite (basic calcium
● Colorless, prism-shaped, co ffin lid appearance, phosphate), Brushite (calcium hydrogen
fern leaf, feathery when disintegrated phosphate)
● Increased in the presence of urea splitting ● Soluble in dilute acetic acid
bacteria
● Soluble in dilute acetic acid
BMLS 3-1 13
ORMAL ALKALINE CRYSTALS
N CHOLESTEROL
CALCIUM CARBONATE ● Rectangular plate with notch in one or more
● Small, colorless, dumbbell, tetrads, spheres corners (staircase pattern)
● Forms gas (effervescence) after adding acetic ● Resembles crystals ofradiographic dye.
acid ● Increased in nephrotic syndrome (lipiduria)
● Misidentified asbacteria ● Soluble in chloroform
How to differentiate your calcium carbonate with your
bacteria?
- Your bacteria, remember, has its characteristic
motility, while your calcium carbonate, being a
crystal, and a nonliving organism, do not have
your motility
RADIOGRAPHIC DYE
● Meglumine diatrizoate, renografin, hypaque
● Flat four-sided plates with notched corner
● Long thin prisms or rectangle
● Resemblescholesterol crystals.
● To differentiate: Check px history; correlatewith
UA results (S.G. >1.040)
BNORMAL ACID CRYSTALS
A - you can check for the patient history
CYSTINE whenever that patient has undergone
● Colorless, refractile, hexagonal plates, often any radiographymethodortesting.And
laminated you can also correlate with other
● Mistaken as uric acid crystals, especially those urinalysis results, such as your specific
uric acid that can form now your hexagonal gravity. If the specific gravity of the
plates patientisgreaterthan1.040,expectthat
● Increased in cystinuria, and cystinosis you will see now your radiographic dye
● Soluble in 10% NaOH
source of error for your cholesterol crystal. Your
radiographic dye
CYSTINE VS URIC ACID
emember, uric acid has a yellow-brown color, while
R
your cysteine is colorless. Both are soluble in ammonia,
and uric acid is insoluble in dilute hydrochloric acid,
while your cysteine is soluble.
BMLS 3-1 14
TYROSINE SULFONAMIDE
● Fine colorless to yellow needles in clumps or ● Fan-shaped, sheaves of wheat, rosettes,
rosettes arrowheads, petals, round shaped, whetstones
● Increased in liver disease more common than ● Mistaken ass calcium phosphate crystal. TO
leucine DIFFERENTIATE: Calcium phosphate: soluble in
● Soluble in alkali and heat acetic acid,Sulfonamide:+LigninTestandDiazo
reaction
- Remember that calcium phosphate is
soluble in acetic acid, and your
sulfonamide has a positive test,
specifically your lining test and your
diazo reaction.
● Increased in possible tubular damage (may
deposit in nephrons)
● Soluble in acetone
BNORMAL ACID CRYSTALS
A
LEUCINE
● Yellow-brown oily looking spheres with
concentric circles and radial striations (sofa
pillow appearance)
● Precipitated with tyrosineafter addingalcohol
● May resemble fat globules
● Increased also inliver disease
● Soluble in alkali or alcohol
AMPICILLIN
● Colorless needles that tend to form bundles
followingrefrigeration
● Increased in massive doses ofpenicillin.
BILIRUBIN
● Clumped granules or needles with brightyellow
color
● Increased in liver disease, OTHER ABNORMAL CRYSTALS (READING ASSIGNMENT)
● Solubleinaceticacid,HCI,NaOH,acetone,and ● HEMOSIDERIN
chloroform ● CYCLOVIR
● INDINAVIR SULFATE
URINE SEDIMENT ARTIFACTS
- These artifacts may be points of errors in your
determination or identification of urinary
sediments.
STARCH GRANULES
● Spheres with dimpled center
● Maltese cross formation under polarizing
microscope
BMLS 3-1 15
POLLEN GRAINS
MALTESE CROSS FORMATION: ● Spheres with cell wall and concentric circles
● Oval Fat Bodies ● Pollen grains are spheres with cell wall and
● Fatty Cast concentric circles.
● Fat Droplets ● May be misidentified as your crystal with your
● Starch concentric circle, and that is your leucine crystal
Commonly, starch granules are seen in the urine when
the registered medical technologist performing the
urinalysis, primarily the preparation of your urine
sediment, has used your latex gloves. Remember, your
latex gloves have starch granules in them which serves
assomewhatalubricantinorderforaregisteredmedical
technologist to properly insert their fingers on the gloves.
OIL DROPLETS
● Mistaken as RBC HAIR FIBERS
● In order to differentiate RBCs from oil droplets, ● Mistaken as casts
you may drop your 2% aceticacid.Whenitwill ● Your hair fibers typically come from your pubic
lyse, it is your RBC hair
VEGETABLE FIBERS
AIR BUBBLES ● Fecal contaminant
● Mistaken as RBC ● Mistaken as casts
● Sameasyouroildroplets,itcanbemistakenalso ● But how can we differentiate casts with your
asyourRBC.Todifferentiateit,youmaydrop2% vegetable fibers as well as your hair fibers?
acetic acid. - the edges of your hair fibers and your
vegetablefibersareclearlydefined,and
it has dark edges. But for the cast,your
casthavinglowrefractiveindex,thecast
havinglowrefractiveindex,theedgesof
your cast may be um somewhat lighter
than the edges of your vegetablefibers
as well as your hair fibers.
BMLS 3-1 16
ACTIVITY 8: QUALITATIVE ANALYSIS OF SUGAR IN URINE ROCEDURE
P
1. Fehling's A (1mL) and Fehling's B (1 mL) in test tube
2. Boil the solution.
A. FEHLING'S TEST 3. While boiling add 1-2 drops ofsample(1mL)every2
● Used to detect the presence of reducing sugars. seconds
● Fehling's A (copper II sulfate) 4. Interpret the result.
● Fehling'sB(Potassiumsodiumtartrateinastrong Mixing Fehling’s A and Fehling’s B will form aDEEPBLUE
alkali, usually sodium hydroxide) COMPLEX. Then boil the solution. WHY? because it
Example of reducing sugars providesthenecessaryheattodrythechemicalreaction
1. Glucose: presence of glucose in the urine is betweenthealdehydeandthefehling’ssolution.Ifthere’s
known asglycosuria no heat there’s no reaction
2. Fructose: Sources of fructose (fruits, vegetables, Why need to add 1-2 drops of sample every 2 seconds?
sweets, desserts) - In order for us to control the reaction rate and
a. Can you find fructose in urine? YES helpthemixturereachaconsistenttemperature.
- Known as fructosuria, it is less common Ensuring it doesn’tproceedtoquickly.Ifnilagay
thanglucose.Itisararegeneticdisorder mo agad lahat ng sample it will overwhelmthe
na tinatawag nating hereditary fructose reagents. It will lead to incomplete reaction or
in tolerance misleading results
3. Galactose: known as galactosuria due to Interpretation
metabolic disorder also known as galactosemia
a. Our body can not properly breakdown
galactose.Whengalactoseaccumulates
it is secreted in our urine.
b. Detecting galactose in our urine is also
crucial for diagnosing of galactosemia,
espen in newborns. WHY? kailangan
magkaroon ng early detection ng
galactosemiasamganewbornbecause
if its not detected earlier it will lead to
serious health issues. For ex. liver
damage, cataracts, intellectual Interpretation
disability, or life threatening infections. Positive Result
HOW? Newborn screening ● Red/orange precipitate: Indicates the presence
4. Maltose:Itisunusuallangtofindmaltoseinurine. of reducing sugars, such as glucoseorfructose.
If maltose is found in urine, pwede siyang mag The bluesolutionofcopper(II)sulfateisreduced
indicate ng carbohydrate metabolism or to a red or orange precipitate of copper(I) oxide.
condition knownasmaltosedeficiencyoritcan Negative Result
also be certain types of glycosuria ● No color change: Indicates the absence of
5. Lactose: Known aslactosuria.Usuallyoccoursin reducing sugars. The blue solution remains
cases of lactose intolerance or during pregnancy unchanged, meaning no reducing sugars are
a. bakit nagkakaroon ng lactosuria during present t o reduce the copper(II) ions.
pregnancy?Mgakatawanngpreganant Bakit pumuputok yung fehling tests? kasi it has copper
women ay nag kakaroon ng metabolic sulfateandsodiumtartrate.Thistworeleasesgasseswhen
changes. So the kidney hindi niya na subjected to higher temperatures
rereabsorb properly yung mga glucose
orlactose.That’swhymaramingglucose B. Trommers’s Method
or lactose na nakikita sa mga ihi ● used to identify water-soluble aldehydes and
because it is not properly absorbed by differentiate them from ketones
the kidneys
PRINCIPLE PRINCIPLE
● Reducing sugars, when heated with Fehling's ● Aldehydes canreducebluecopper(II)sulfateto
solution,reducethebluecopper(II)ionstoredor red or orange copper(I) oxide in an alkaline
orange copper(I)oxide,indicatingthepresence solution, indicating the presence of reducing
of reducing sugars. sugars or other aldehyde-containing
compounds.
BMLS 3-1 17
uestion: Is aldehyde considered as reducing sugars?
Q RINCIPLE
P
NO,butsomereducingsugarscontainsaldehydegroups Reducing sugars, like glucose and fructose,canreduce
and ang tawag sa kanila ay aldolases (ex. GLUCOSE) blue copper(II) sulfate to red copper(I) oxide in an
alkaline solution, causing a color change.
rocedure
P
1. 2 mL of sample in test tube PROCEDURE
2. Add 1 mL 10% KOH in test tube 1. Put 3 mL of Benedict's reagent in a test tube.
3. Add 5 drops of 10% copper sulfate 2. Add 5-8 drops of sample.
4. Boil - bakit 5-8 lang? because it gives a precise
5. Interpret balance, if masyadong marami it will lead to
What is the relevance of adding 1 mL 10% KOH in test dilution
tube? 3. Mix well and boil for 2-5 minutes.
- tocreateaalkalineenvironment.TheKOHhelps 4. Cool in tap water
to maintains the stability of copper hydroxide - whydoweneedtocoolitinatapwater?Tostop
precipitate which is the active reagent of the reaction
trommers tests 5. Note the color change and interpret.
Ano ang ginagawa ng adding 5 drops of 10% copper
sulfate
- it providesthenecessaryamountofcopperions INETERPRETATION
for the reaction. Yung mga ions kasi sila yung
mga nag rereact sa mga reducing sugars na
meron sa sample
Interpretation
C. Benedict’s Test
Interpretation
Positive Result
Formation of a red or orange precipitate
indicates the presence of an aldehyde or a
reducing sugar
Negative Result
No color change or precipitate formation
indicates the absence of an aldehyde or a
reducing sugar INTERPRETATION
C. Benedict's Test LUE- NEGATIVE FOR REDUCING SUGAR
B
- used to detect the presence of reducing sugars YELLOW/GREEN- TRACE AMOUNT OF REDUCING SUGAR
1. Copper(II) sulfate (CuSO4): Provides the ORANGE- MODERATE AMOUNT OF REDUCING SUGAR
copper(II) ions that react with reducing sugars. BRICK RED- LARGE AMOUNT OF REDUCING SUGAR
2. Sodium carbonate (Na2CO3): Creates an
alkaline environment necessary for the reaction.
3. Sodium citrate (Na3C6H507): Acts as a
complexing agent to keep the copper ions in
solution.
BMLS 3-1 18
THER QUALITATIVE TESTS FOR REDUCING
O
SUGARS
● T ollens' Test: Involves ammoniacal silver nitrate,
where reducing sugars reduce silver ions to
metallic silver, forming a mirror-like coating.
● arfoed's
B Test: Differentiates between
monosaccharides and disaccharides using
copper(II)acetateinanacidicmedium,forming
a red precipit ate with monosaccharides.
● Ifthesolutionremainebluethencarbohydrateis
present, then if red precipitate is formed faster,
monosaccharides is present, then if its formed
slower then disaccharides is presnet
● NS Test (Dinitrosalicylic Acid Test): Reacts with
D
reducing sugars to for m a colored complex,
changing from yellow to red.
● linitest: Utilizes a tablet format for detecting
C
glucoseinurine,resultinginacolorchangefrom
blue to orange.
BMLS 3-1 19
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