Automated urine analysis systems can perform a complete urinalysis including physical, chemical, and microscopic components. They provide standardized, efficient results with improved turnaround times compared to manual methods. Key advantages include objective particle identification and classification through techniques like flow cytometry and digital image analysis, as well as automated test strip reading and sample handling for tests like glucose, protein, blood, and more. While providing productivity and consistency benefits, automated urine analysis aims to enhance urine sediment examination and reporting.

1. AUTOMATED URINEANALYSIS
Presenter : Dr Manjunatha T M
Moderator: Dr. Rajashekhar. K.S

2. The history of urinalysis
The ancient world
The origin of visual urine diagnostics, can be traced back to
ancient Egypt.
Hippocrates (approx. 400 BC) recognized that
urine characteristics (odor / color) were altered with different
diseases.

3.  Six centuries later, Galen (AD 129–200) refined Hippocrates
ideas, theorizing that urine represented is not a filtrate of the
four humors and but rather, a filtrate of the blood . An
increasing number of physicians were diagnosing from urine
alone.
 Amateurs (called‚leches‘) started diagnosing based only on the
color of urine.

4.  The first “test strips” were developed by the Parisian
chemist Jules Maumene (1818–1898) when, in 1850,
he impregnated a strip of merino wool with “tin
protochloride” (stannous chloride).
 On application of a drop of urine and heating over a
candle the strip immediately turned black if the urine

5. contained sugar and it took another 70 years before the
Viennese chemist Fritz Feigl (1891–1971) published his
technique of “spot analysis.”

6.  Urine test strips in the sense used today were first made
on industrial scale and offered commercially in the 1950s.
 The company Boehringer Mannheim, today a top
leader on the world market under the name of Roche,
launched its first Combur- TestR strips in 1964.

7. Urine analysis is a valuable tool used to diagnose and
monitor renal and urinary tract illnesses.
Typically it is a moderate to high sample volume test for a
general chemistry lab, representing up to 30 % of all samples
received .
Routine urinalysis consists of macroscopic examination,
chemical analysis and microscopic urine sediment examination.

8. Urinalysis Automation
Several automated instruments are currently available to
standardize:
• Sample processing
• Biochemical test strips analysis
• Microscopy analysis
• Report results
8

9. 9
Automation Urinalysis Features:
1. On-line computer capability
2. Bar coding
3. Manual entry of color
4. Clarity
5. Microscopic results

10. There are three ways to perform a test strip analysis:
• Manual – The test is done by hand
• Semi-automated – The test strip is dipped in the urine
manually and then analyzed by an instrument
• Fully-automated – The test strip is analyzed completely
by an instrument

11.  Automated urine cell analyzers mix, aspirate, dilute and stain
urine to classify urine sediment particles.
 Automated urine systems perform a complete urinalysis that
includes the physical, chemical and microscopic parts of a
routine

12. Urinalysis Automation
EquipmEnt manufacturE
1) Semiautomated Chemistry Instruments
2) Clinitek 200/200 + Siemens Medical Diagnostics
3) Clinitek 500 Siemens Medical Diagnostics
4) Chemstrip Urine Analyzer Roche Diagnostics
5) Urisys 1800 System Roche Diagnostics

13. 2) Fullyautomated Chemistry Instruments
I.Clinitek Atlas Siemens Diagnostics
II.Chemstrip Super Automated
Urine Analyser Roche
Diagnostics
bl.Urisys 2400 system Roche
Diagnostics
3) Automated Microscopy
I.UF-100 Urine Cell Analyser Sysmex
II.iQ200 Automated Urine Microscopy Iris Diagnostics Division

14. 4) Automated Urinalysis Systems
I.ADVIA Urinalysis Workcell Siemens Diagnostics
II.iQ200 Automated Urinalysis
System Iris Diagnostics Division

15. Clinitek 50/100
15

16. Clinitek 500
• Distinguishes between
hemolyzed and nonhemolyzed
specimen
• Determine low SG and pH
• Rapid entry
• Specimen ID
• Color
• Clarity
• Automatic features
• Color determination
• Strips detection
• Calibration
• Confirmatory
• Microscopic analysis
16

17. Clinitek Status Analyzer
• The Analyzer is for in vitro use in the semi-quantitative detection of
Albumin, bilirubin, blood (occult), creatinine, glucose, ketone
(acetoacetic acid), leukocytes, nitrite, pH, protein, specific gravity and
urobilinogen in urine samples
• The calculation of albumin-to-creatinine and protein-to-creatinine ratios in
urine samples, when Clinitek® Microalbumin and Multistix PRO®
Reagent Strips for Urinalysis are used
• The detection of human Chorionic Gonadotropin (hCG) in urine samples,
when Clinitest® hCG cassettes are used 17

18.  Urine Reagent Strips are made for urinalysis of both qualitative
and semi-quantitative, which are in vitro reagent for
diagnostics.
 The results on the strips can be read visually and
instrumentally. The pH and Protein can be read at any
time within 60 seconds after dipping.
 For a qualitative result, the strip should be read between 1-2
minutes after dipping .Colour changes beyond 2 minutes are of
no diagnostic value.

19. Reaction Principles
1)Glucose: The glucose oxidized by glucose oxidase catalyzes the
formation of glucuronic acid and peroxide hydrogen. Peroxide hydrogen
releases oxide (0) under the function of peroxidase. (0) oxidizes Iodide
potassium, which makes the colour change.
2)Bilirubin: The direct bilirubin and dichlorobenzene diazonium produce
azo dyes in a strongly acid medium.
3)Ketone: The acetoacetic acid and sodium nitroprusside cause reaction
in the alkaline medium, which produces a violet colour.

20. 4) Specific Gravity: Electrolyte (M X) in the form of salt in urine
reacts with poly methyl vinyl ether and malefic acid (-COOH), which are
weak acid ionic exchangers. The reaction produces hydrogenous
ionogen, which reacts with pH indicator that causes the colour change.
5) Blood: Haemoglobin acts as peroxidase. It can cause peroxide
release, neo-ecotypes oxide (O) oxidizes the indicator and makes the
colour change subsequently.

21. 6) pH: The method of the pH indicator is applied.
7) Protein: This is based on the protein-error-of-indicator principle.
Anion in the specific pH indicator attracted by caution on the protein
molecule makes the indicator further Ionized, which changes its colour.
8) Urobilinogen: Urobilinogen and diazonium produce pink azo dyes
under the function of a strong acid medium.
9) Nitrite: Nitrite in the urine and aromatic amino sulphanilamide are
diazotized to form a diazonium compound. The diazonium compound

22. reacting with tetrahydro benzo (h) quinolinphenol causes the colour
change.
10 )Leukocytes: Granulocyte leukocytes in urine contain esterase’s
that catalyze the hydrolysis of the pyrrole amino acid ester to
liberate 3-hydroxy 5-pheny pyrrole. This pyrrole reacting with
diazonium forms a purple colour.

23. Reactive ingredients (based on dry weight at time of impregnation)
oProtein: 0.1% w/w tetrabromphenol blue; 97.4% w/w buffer: 2.5/) w/w non reactive
ingredients.
oBlood: 26.0% w/w diisopropylbenzene dihydro peroxide; 1.5% w/w
tetramethylbenzidine; 35.3% w/w buffer; 37.2 % w/w non reactive ingredients.
oGlucose: 1.7% w/w glucose oxidase (microbial 123U); 0.2% w/w peroxidase
(horseradish. 203 IU); 0.1 % w/w potassium iodide; 71.8% w/w buffer; 26.2% w/w non
reactive ingredients.
oKetone: 5.7% w/w sodium nitroprusside; 29.9% w/w non reactive ingredients:
64.4% w/w buffer.
oLeukocytes: 4.3% w/w pyrrole amino acid: 0.4% w/w diazonium salt; 92.6% w/w
buffer, 2.7% w/w non reactive ingredients.

24. o Nitrite: 1.3% w/w p-arsanilic acid; 0.9% w/w tetrahydroquinoline N-(1-
Naphthol)-ethylenediamine; 89.6%, w/w buffer; 8.2% w/w non
o reactive ingredients.
o Specific Gravity: 4.8% w/w bromthymol blue; 90.2% w/w poly(methyl vinyl
ether co maleic anhydride); 5.0% w/w sodium hydroxide.
o pH: 3.3% w/w bromcresol green; 55.0% w/w bromthymol blue: 41.7% w/w
non reactive ingredients.
o Bilirubin: 0.6% w/w 2.4 -dichlorobenzene amine diazonium salt; 57.3% w/w
buffer; 42.1% w/w non reactive ingredients.
o Urobilinogen: 0.2& w/w fast blue B salt; 98.0% w/w buffer; 1.8% w/w non
reactive ingredients.

25. Siemens Clinitek Microalbumin 2 Reagent Strips
• Provide albumin, creatinine, and albumin to creatinine ratio results
in 1 minute
• useful to test for microalbuminuria in patients with diabetes
or hypertension in order to detect early kidney disease
• Use with Clinitek 50 or Clinitek Status analyzers
– Sensitivity as low as 2mg/dL for urine protein
– More reliable; less affected by interferences (e.g. specific gravity
and pH)

26. Siemens Clinitek Microalbumin 9 Reagent Strips
• Provide albumin, blood, creatinine, glucose, ketone, leukocyte,
nitrite, pH, & protein and albumin to creatinine ratio & protein to
creatinine ratio
• Use with Clinitek Status or Advantis analyzers
– Random sample; no timed or 24 hr urine sample required
– Accurate identification of microalbuminuria

27. Dirui H-500 Urine Analyzer

28.  The Dirui H-500 Urine Analyzer provides results for urine
testsamples based on an advanced high luminosity cold light
source with 4-wavelength technology. This improves sensitivity,
accuracy and specificity and reduces ambient light interference.
 Automatic waste handling avoids sample cross-contamination and
the analyzer offers a quiet, high speed built-in thermal printer or
external stylus printer

29. Additional Information about the H-500
Test wavelength: 525nm, 572nm, 610nm, 660nm
Offers a test throughput of 514 strips per hour
Provides a data memory of 5,000 patient results
A 5.7″ LCD display provides ease of use
Comes with a built-in high speed, low noise thermal printer
Continuous feed
Flag abnormal values
Test Items
Urobilinogen, Bilirubin, Ketone, Blood, Protein, Nitrite,
Leukocytes, Glucose, Specific gravity, pH

30. + Urobilinogen
+ Bilirubin
+ Ketone
+Blood
+Protein
- Nitrite
++ Leucocytes
+++ Glucose
1.020 SG
7.0 pH
Used Strip Fresh Strip

31. URISYS 2400 system Chemstrip super automated

32. • Technique
Reflectance Photometry - uses the principle that light reflection
from the test pads decreases in proportion to the intensity of
color produced by the concentration of the test substance.
A monochromatic light source is directed toward the reagent
pads.

33.  The light is reflected to a photodetector and an analog
/digital converter.
 The ultimate goal of automation is to improve reproducibility
and color discrimination, increasing productivity and
standardization for reporting urinalysis results

35.  The Chemstrip Super Automated Urine Analyser and the
Roche Diagnostics Urisys 2400 system are fully automated
‘walk-away’ urine chemistry instruments for a large urinalysis
laboratory.
 With the Chemstrip Super Automated Urine Analyser, sample
volumes are detected, adjusted and automatically mixed.

36. • A sorter mechanism supplies a single test strip from the
sorter drum to a sorter position.
• A gripping mechanism grasps the test strip and dips it in to
the urine specimen tube.

37. • The dipping mechanism lifts the test strip out of the
sample tube while removing excess urine by dragging
the strip along the inside of the specimen tube
• The dipping mechanism then transfers the test strip to
the Reflectance Photometer position.

38. • A transport plate positions the test strip at the Reflectance
Photometer recording head, where specimen is measured at
three different wavelengths (555,620,660 nm) at 48 seconds and
120 seconds after dipping.

39. • The Urisys 2400 system utilizes a pippetting unit that
automatically mixes the specimen and pipettes the precise
volume to each test pad.
• The minimum sample volume is 1.5 ml. Four hundred test
strips are loaded into a Urisys 2400 cassette and the strips
are stable in the cassette for 2 weeks. 75 samples per load
for immediate measurement of emergency samples.

40. • It designed for large Lab
• Performs >12 tests automatically
• Walk-away capability (> 225
specimen/hr)
• > 2 mL urine specimen required
• Flagging abnormal
specimen
• Automatic features
• Color determination
• Strips detection
• Calibration
• Confirmatory
• Microscopic analysis
• etc
Clinitek AtlasClinitek Atlas

41. Automated Microscopy
Automated Urine cell analyzers provide efficient standardized
results in about a minute, markedly improving turnaround
times.
The Sysmex UF – Series offers fully automated sample
analysis with automatic classification of all 10 formed element
groups with scattergrams and histograms for reference.

42. Disadvantages of Manual Urine Sediment
Microscopy
o Subjective element identification
o Poor reproducibility
o Lack of standardization
o Time consuming/labor intensive

43. • The UF-100 and UF –50 use laser – based flow cytometry
along with impedance detection, forward light scatter and
fluorescence to identify the individual characteristics and
stained urine sediment particles in a flowing stream.

44. Sysmex UF 50 Sysmex UF 100

45. The sample is stained with two dyes that radiate an orange
and green fluorescence. The DNA within the cells is stained
by the orange dye, phenathridine ; the nuclear membranes,
mitochondria and negatively charged cell membranes are
stained with a green dye, carbocyanine
The stained sample is passed through the flow cell,
presented to a laser light beam that produces fluorescence
and light scatter

46. • The main parameters are RBCs, WBCs, epithelial cells, casts and
bacteria. Flagging parameters include pathologic casts, crystals,
small round cells, sperm and yeast like cells.
• Particles are identified by measuring the change in
impedance of the sediment elements, as well as the height
and width of the fluorescent and light scatter signals,
presented in scattergrams and histograms.

49. IQ 200 Automated Urine microscopy analyzer
 Automatically analyzes and Classifies urine particles in to 12
categories. The sample is mixed and aspirated to a planar
flowcell where 500 digital photomicroscopic images are
taken per sample.

50.  The system uses Auto Particle Recognition (APR) software that
Classifies urine particles in the photographs based on size, shape,
texture and contrast in to 12 categories – RBCs, WBCs, WBC clumps,
hyaline casts, unclassified casts, squamous and non Squamous
epithelial cells, bacteria, yeast, crystals, mucus and sperm

52. IRIS Flow Videomicroscopy
• Urine is drawn through a flat
chamber
• Video snaps are sorted by
computer
• Technician scans images and
deletes dud ones
Computer then adds up #/cmm
• These are RBCs
52

53. IRIS Flow Videomicroscopy
• Squamous epithelial cells
53

54. Accuracy of the iQ200 and UF-100 systems in comparison
with microscopic results.
Parameter Accuracy (95% CI), %
iQ200 UF-100
Leukocytes 89 (85.5–92.5)
84 (80–88)
Erythrocytes 86 (82–90)
81 (77–85)
Bacteria 68 (63–73)
42 (36.5–47.5)
Pathological casts 91 (88–94)
86 (82–90)
Yeasts 97 (95–99)
93 (90–96)
Crystals 92 (89–95)
88 (84–92)

55. Sensitivity and specificity of the systems calculated
based on the cut-off values.
iQ200 Sensitivity % Specifity % NPV % PPV
%
Leukocytes 76 97.5 94
89
Erythrocytes 70 98 90 92
Bacteria 85 95 88
94
Pathological 68 97 93
83
casts
Yeasts 70 99 97
91
Crystals 71 97 95
80
UF-100
Leukocytes 92 90 98

56. Automated Urinalysis Systems
• Clinitek Atlas, an automated urine chemistry analyzer and the
Sysmex UF-100, an automated urine cell analyzer, have been
integrated to develop the ADVIA Urinalysis Workcell System.

57. Sysmex UF-1000i

58. Sysmex UF-1000i
• Laser-based flow cytometer utilizing 2 stains with fluorescent
dyes to stain cellular elements
• Separate bacteria channel for improved discrimination
• Forward scatter, hydrodynamic focusing, forward fluorescent
light, conductivity measurements, and adaptive cluster
analysis

59. Sysmex UF-1000i System Components
• Main unit with integrated pneumatic unit
• IPU (information processing unit) Windows XP operating system
• Sampler unit with tube rotator unit
• Bar code reader
• Laser Jet graphic printer/line printer (1 device, 2 settings)
• Handheld bar code reader

60. The Sysmex® UF-1000i, an automated urine particle analyzer, is a
dedicated system for the analysis of microscopic formed elements in
urine specimens. The instrument consists for three principal units:
(1)Main Unit which aspirates, dilutes, mixes and analyzes urine
samples;
(2) Auto Sampler Unit supplies samples to the Main Unit
automatically;

61. (3) IPU (Information Processing Unit) which processes data from
the Main Unit and provides the operator interface with the system.
The UF-1000i is equipped with a Sampler that provides continuous
automated sampling for up to 50 tubes.

62. • The instrument utilizes Sysmex flow cytometry using a red
semiconductor laser for analyzing organized elements of urine.
• Particle characterization and identification is based on
detection of forward scatter, fluorescence and adaptive cluster
analysis.
• Using its own reagents, the UF-I000i automatically classifies
organized elements of urine and carries out all processes
automatically from aspiration of the sample to outputting the
results.

63. UF-1000i Tube Rotator

64. UF-1000i Reagents
UFII SEARCH™-SED
UFII PACK™-SED
UFII SHEATH™
UFII SEARCH™ -BAC
UFII PACK™-BAC

65. UF II PACK-SED / UF II SEARCH-SED
 UF II PACK-SED
Removal of amorphous salts together with heating (up to 35°C)
 UF II SEARCH-SED
Polymethine dye
Chromogen chain with electron donor and acceptor group
Stains parts of nucleus, parts of cytoplasm and membranes
Excitation wavelength is 635 nm
Emission wavelength is over 660 nm

66. UF II PACK-BAC / UF II SEARCH-BAC
• UF II PACK-BAC
– UF II PACK-BAC (e.g. its pH value) together with heating to >40°C
suppresses non-specific staining of particles other than bacteria
• UF II SEARCH-BAC
– Polymethine dye
– Distinctively stains nucleic acid elements in bacteria

67. Flow cell
Laser light
Laminar Flow
particles
Sheath reagent
Sheath
nozzle
Scattered
light

72. UF-1000i Detection Parameters
MucusBacteria
Sperm
YeastHyaline Casts
Small Round CellsEpithelial Cells
CrystalsWBC
Pathological CastsRBC
Flagged ParametersEnumerated Parameters

73. RBC
Small - medium size
Low fluorescenceFl
Fsc
Fl
Fsc
Medium - large size
Medium - high
fluorescence
Fl
Fsc
Very small size
Size(sectionalarea)
Large
Small
Fluorescence HighLow
Bacteria
WBC
Low fluorescence
S_FLH
S_Fsc
S1: FLH / Fsc - Scattergram
YLC
X’TAL
Sperm
Low to medium fluorescenceFl
Fsc
Small size
Medium fluorescenceFl
Fsc
Small size
Fl
Fsc
Small - large size
no fluorescence

74. YLC
RBC
Small - medium size
Low fluorescenceFl
Fsc
Fl
Fsc
Fl
Fsc
Size(sectionalarea)
Large
Small
Fluorescence HighLow
Bacteria
WBC
Low fluorescence
S_FLL
S_Fsc
S2: FLL / Fsc - Scattergram
Low to medium fluorescence
Fl
Fsc Small size
Medium fluorescenceFl
Fsc
Small size
Fl
Fsc
Medium – very large size
Medium - high fluorescence EC
Sperm
Very small size
Medium - high fluorescence
Medium - large size

75. S3: FLLW2 / FLLW - Scattergram
FLLW2
S_FLLW2
Large
Small
Length of stained particleShort S_FLLW
FLLW
FLLW2
Little to more stainable inclusions
FLLW
FLLW
Long
Length of stained inclusions
Casts (no inclusions)
Lengthofstainedinclusions
Short –
medium length
of inclusions No to little inclusions
More stainable inclusions
Path.
casts
Epithelial cells
Mucus
FLLW2
FLLW
No inclusions
SRC
WBC
FLLW2
Long

76. B1: Fsc / FLH - Scattergram
B_FSC
Large
Small
Stainability of particlesLow B_FLH
Sizeofparticles
BACTDebris
Weak fluorescenceFlH
Fsc
Small size
FSC
FLH
Small to big size
No fluorescence
High

77. UF-1000i
Sediment 1

78. UF-1000i
Sediment 2

79. UF-1000i
Sediment 3

80. UF-1000i
Bacteria 1

81. UF-1000i
Bacteria 2

82. UF-1000i
Bacteria 3

83. UF-1000i Technology
BacteriaSediments
BacteriaSediments Stain
IncubationIncubation
DetectionDetection
unitunit
Sediments Bacteria
Diluents
Two chambers for stain and dilution
Improved determination of
bacteria
Red semiconductor laser
•Down sizing
•Long life
•Reduced power consumption

84. Sample Incubation
• Incubation time at certain temperature ranges needed
for staining
– for the SED analysis:
• 10 seconds at 35°
C
– for the BAC analysis:
• 20 seconds at 42°
C

85. UF-1000i Technology
Fluorescence
Stain DNA/RNA
Non-specific staining with debris
ForwardScatter
Specific stain for Nucleic AcidSpecific stain for Nucleic Acid
Dye
Dye
Dye
Dye
Dye
Dye
Dye
DyeDye
Dye
Dye
Dye
Dye
Dye
Dye
Dye
Dye
Dye
Dye
DyeDye
Dye
Dye
Dye
DyeDyeDye
Dye
1) Enhanced detection of
bacteria
2) Staining bacteria
nuclei
Polymethine dye

86. UF-1000i Technology
UF-100 :
RBC 119.8/µL
X’TAL 0.0 /µL
UF-1000i: RBC 3.3/µL
X’TAL 102.7/µL
Microscopy :
RBC 5.6/µL
X’TAL (2+)
false-positive by X’TAL interferenceReduction of false-positive by X’TAL interference to RBCScattergram
UF-100UF-1000i
The more complex the
surface or inner
construction, the more
intensive SSC signal is.
S-FSC

87. UF-1000i Technology
UF-100 :
EC 83.4/µL
UF-1000i: EC 18.2/µL
Microscopy:
EC 24.5/µL
WBC cluster can be detected as
EC. It is false positive of EC.
Reduced false positive EC
with high positive WBC
Scattergram
UF-100UF-1000i
SSC parameters can help UF to
distinguish WBC and EC.
WBC is accurately classified by SSC
signals.

88. References
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clinical laboratory Part 1—Ancient times through the 19th century
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http://www.academia.dk/Blog/wp-content/uploads/KlinLab-Hist/LabHistory1.pdf
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Pincus MR, eds. Henry’s Clinical Diagnosis and Management by
Laboratory Methods. 23rd Ed. China: Elsevier; 2017. p. 441–80.
Richard Thompson, Andrew Gammie, Debbie Lewis, Rebecca Smith
CE. Evidence review: Automated urine screening systems. CEP10030.
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urinalysis: First experiences and a comparison between the Iris iQ200
urine microscopy system, the Sysmex UF-100 flow cytometer and
manual microscopic particle counting. Clin Chem Lab Med [Internet].
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