Disclaimer: The notes I’m sharing may not be currently updated (as I’ve used some of them in my past lectures/reviews), so please always refer to textbooks if you encounter conflicting information.
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ALL NOTES MAY BE USED FOR UNDERGRADUATE, LOCAL BOARD EXAM OR INTERNATIONAL EXAMS (i.e., ASCPi)
— This list will be updated from time-to-time, so check back regularly. Click DOWNLOAD BUTTON to access each file below. —
ERRATUM FOR THIS:For the action of insulin, please change GLYCOGENOLYSIS to GLYCOGENESIS. Insulin lowers glucose and so it should increase conversion of glucose to glycogen (GLYCOGENESIS).
Glucose (correlated with ketones) Double sequential enzyme reaction
ENZYMES: Glucose oxidase Peroxidase
30 s
M – glucose oxidase, peroxidase, potassium iodide C – glucose oxidase, peroxidase, tetramethylbenzidine
CHROMOGENS: O-toluidine (pink to purple) Potassium iodide (blue to brown) Aminopropryl-carbazol (yellow to orange-brown) Tetramethylbenzidine (yellow to green)
Contamination by oxidizing agents and detergents
High levels of ascorbic acid, ketones, specific gravity Low temperature Improperly preserved specimens
Ketones (correlated with glucose)
Sodium nitroprusside reaction
40 s
M – sodium nitroprusside (acetoacetic acid)
C – sodium nitroprusside + glycine (acetoacetic acid & acetone)
Phthalein dyes, highly pigmented red urine, levodopa Medications containing free sulfhydryl groups (MESNA)
Improperly preserved specimens
Specific Gravity
pKa change of polyelectrolyte
pKa = dissociation constant
45 s
M – poly (methy lvinyl ether/maleic anhydride) bromthymol blue
C – ethyleneglycol-Bis (aminoethylether) bromthymol blue
High concentrations of proteins because of protein anions
Highly alkaline urines (>6.5) Add 0.005 to S.G. readings
pH (correlated with Nitrite, LE, microscopic)
Double-indicator system
60 s
Methyl red Bromthymol blue
None
Runover from the adjacent CHON pad may lower pH
Protein (correlated with blood, nitrite, LE, microscopic)
Protein error of indicators
60 s
M – tetrabromphenol blue
C – tetrachloropenol tetrabromosulfonphthalein
ACID BUFFER: Citrate
Highly buffered alkaline urine High specific gravity pigmented specimens, phenazopyridine quaternary ammonium compounds (detergents) antiseptics, chlorhexidine loss of buffer from prolonged exposure of reagent strip to the specimen
Proteins other than albumin
Blood (correlated with protein and microscopic)
Pseudo-peroxidase activity of hemoglobin
60 s
M – diisopropylbenzenedehydroperoxide tetramethylbenzidine
C – dimethyldihyroperoxide- tetramethylbenzidine
spotted blue for intact RBCsuniform blue for Hb and myoglobin
C – old specimens, preservation in formalin, high concentration of nitrite
Nitrite (correlated with protein, LE and microscopic)
Greiss reaction
60 s
M – p-arsanilic acid Tetrahydrobenzo(h)-quinolin-3-ol
C – Sulfanilamide, hydroxytetrahydro benzoquinoline
Improperly preserved specimens
Highly pigmented urine
Nonreductase-containing bacteria insufficient contact between bacteria and urinary nitrate (should be 4 hours) lack of urinary nitrate, large quantities of bacteria converting nitrite to nitrogen, presence of antibiotics, high concentration of ascorbic acid high specific gravity
Leukocytes (correlated with protein, nitrite and microscopic)
Leukocyte esterase
120 s
M – derivatized pyerole amino acid ester, diazonium salt
Principles Used in Automated Urinalysis and Microscopy
Automated Urinalysis Systems
Automated Body Fluid Analysis Systems
Principles Used in Automated Urinalysis and Microscopy
REFLECTANCE PHOTOMETRY
Used by automated reagent strip readers
Measure the light reflected from the reagent strip color pads and compare the amount of reflected light with a known standard
PRINCIPLE: light reflected from the colored reagent pads DECREASES in DIRECT PROPORTION to the INTENSITY OF THE COLOR produced by the reaction with the specific substance in the urine sample
the darker the color, the less light reflected
the lighter the color, the more light reflected
The concentration of a specific substance and concentration units are displayed on the reader’s display
Conductivity is based upon the impedance (the amount of resistance that occurs when a sediment passes through an electrical field) of sediments and counts the numbers of pulses (sediments).
The size of the pulse indicates the size of the sediment.
Light scattering characteristics of the sediments are determined by their movement through the laser light beam.
Identification depends on how the light is scattered by the sediment.
HARMONIC OSCILLATION
Assesses SPECIFIC GRAVITY
Method based upon densitometry in which a sound wave of a specific frequency changes in proportion to the density of the urine sample
Change in wave frequency is measured by a microprocessor and translates the reading to specific gravity
HYDRODYNAMIC FOCUSING
Identifies specific sediments
involves the movement of single urine sediments past the optics of a microscope to allow sediments to flow in several planes plane past the microscope objective
A flow cell also measures sediment conductivity, size, and light scattering traits
Questionable findings are viewed on a monitor for operator identification and confirmation
Automated Urinalysis Systems
INDIVIDUAL STRIP READERS
SEMIAUTOMATED ANALYZERS
dependent on an operator for specimen mixing, test strip, dipping, and inputting of physical and microscopic results
FULLY AUTOMATED CHEMISTRY ANALYZERS
add urine to the reagent strip
AUTOMATED URINE CELL ANALYZERS
mix, aspirate, dilute, and stain urine to classify urine sediment particles
COMPLETELY AUTOMATED SYSTEMS
perform a complete urinalysis that includes the physical, chemical, and microscopic parts of a routine urinalysis
WAIVED URINE CHEMISTRY INSTRUMENTS
Roche Diagnostics Criterion II Siemens Medical Solutions Diagnostics Clinitek®50 Siemens Medical Solutions Diagnostics Clinitek®101 Siemens Medical Solutions Diagnostics Clinitek®Status
Iris Diagnostics Division iQ®200 Automated Urinalysis System iRICELL2000 (iChem® VelocityTM plus iQ®200ELITETM) iRICELL3000 (iChem® VelocityTM plus iQ®200SPRINTTM) Siemens Medical Solutions Diagnostics ADVIA Urinalysis WorkCell System (Clinitek® Atlas plus the Sysmex UF-100)
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DESCRIPTION OF SOME SELECTED INSTRUMENTS
CLINITEK 50 & CLINITEK STATUS
well suited for small volume laboratories and physician’s offices
Memory storage for test results -100 for Clinitek 50 and 200 for Clinitek Status
automated reading of microalbumin-to-creatinine and protein-to-creatinine ratios and human chorionic gonadotropin (hCG)
CLINITEK 200
For medium-volume to large-volume urinalysis laboratories and features a high specimen output of one strip every 10 seconds.
Multistix reagent test strips are used, and the instrument has the ability to report semiquantitative (mg/dL) results or plus (+) and SI units.
The reflectometer is calibrated daily and maintenance is required each day for all areas in contact with urine test strips
SYSMEX UF-SERIES
Fully automated sample analysis with automatic classification of all 10 formed element groups with SCATTERGRAMS and HISTOGRAMS for reference
Laser-based FLOW CYTOMETRY along with impedance detection, forward light scatter, and fluorescence
Sample is stained with 2 dyes
PHENATHRIDINE – orange dye, stains DNA
CARBOCYANINE – green dye, stains nuclear membranes, mitochondria, and negatively charged cell membranes
SYSMEX UF-SERIES
Stained sample is passed through the flow cell, where it is HYDRODYNAMICALLY FOCUSED and presented to a laser light beam that produces fluorescence and light scatter
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, which are presented in scattergrams and histograms
iQ 200 Automated Urine Microscopy Analyzer (IRIS)
Automatically analyses and classifies urine particles into 12 categories
Uses AUTO PARTICLE RECOGNITION (APR) software that classifies urine particles in the photographs based on size, shape, texture, and contrast
Automated Body Fluid Analysis Systems
cells are first mixed with reagent fixative and then counted
differentials countingenumerates numbers of neutrophils, lymphocytes, monocytes, and eosinophils
automated cell counters use larger numbers of cells, enhancing precision and accuracy
Siemens Medical Solutions Diagnostics ADVIA120 and 2120
CSF
Sysmex XE-5000 Automated Hematology System
CSF Serous body fluids Synovial fluid
Medical Electronic Systems
Semen
ADVIA120 Hematology System
First automated instrument with an FDA-approved automated CSF assay
Uses flow cytometry, light scatter, and absorbance to count the RBCs, WBCs, and performs a WBC differential that includes percentages and absolute numbers of mononuclear cells and PMNs on samples with >20WBCs/µL
AUTOMATION OF SEMEN ANALYSIS
SQA-V automates sperm counts and motility
has a two-channel measurement system that interacts with a specially designed testing capillary that contains the semen sample
one channel “measures” light absorption and refraction in sperm cells and translates this into concentration
one channel “counts” light interruptions (signals) caused by sperm cells moving across the field of light
In approximately 1 minute, thousands of signals are “read” resulting in exceptional accuracy and precision.
Automating the motility analysis eliminates reader subjectivity and variance among technologists.
AUTOMATION OF URINE PREGNANCY
Quantitative human chorionic gonadotropin (HCG) is one such test that is interpreted by the VEDALAB Easy Reader.
Immunochromatographic rapid test cards are read by the meter using a high-resolution CCD camera.
Integrated software analyzes the images and records the results.
My son, Ily, endorses Hematology as your study companion! ❤
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GCASH If GCASH to GCASH or GCASH to bank, no extra fee If kiosk (i.e., 7-11, palawan, cebuana) to gcash, please add the required service fee to the amount to be transferred PALAWAN or CEBUANA to GCASH: add 2% service fee 7-11 to GCASH: add 3% service fee
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Doc Krizza-Almond 