Instrumentation, Principles of

Instruments, and Spectrophotometry

Light Energy, Wavelength, and Radiant Energy

● Energy is transmitted via electromagnetic waves that are characterized by

their frequency and wavelength.
● Wavelength is the distance between two successive peaks and it is
expressed in nanometer (nm).
● Nominal wavelength represents the wavelength in nanometer at peak
transmittance.
● A slight error in wavelength adjustments can introduce a significant error in
absorbance readings.
● The relationship between wavelength and energy (E) is described by Planck's
formula.

Wavelength Regions
● Visible spectrum: 400 - 700 nm
● Ultraviolet region (UV): <400 nm
● Infrared region (IR): >700 nm

Planck's formula: E = hv
where:
E - is the energy of a photon in Joules
h - constant (6.626 x 10-27 erg sec) → Bishop
v - frequency

Frequency
● Frequency is the number of vibrations of wave motion per second.
● The lower the wave frequency, the longer the wavelength.
● The wavelength is inversely related to frequency and energy; the shorter the
wavelength, the higher the frequency and energy, and vice versa.

Single-beam Spectrophotometer
Major Analytical Methods in the Clinical Chemistry ● It is the simplest type of an absorption spectrophotometer.
Laboratory ● It is designed to make one measurement at a time at one specified
wavelength.
I. COLORIMETRY ● The maximum absorption of the analyte must be known in
● Spectrophotometric measurement - is the measurement of light intensity in advance when a single-beam instrument is used.
a narrower wavelength.
● Photometric measurement - is the measurement of light intensity using a
specific wavelength
● The primary analytical utility of spectrophotometry or filter photometry is
the isolation of discrete portions of the spectrum for purposes of
measurement.

Spectrophotometry
● It involves measurement of the light transmitted by a solution to
determine the concentration of the light-absorbing substances in
the solution.
Double-beam Spectrophotometer
● A spectrometer is a device that measures the wavelengths of light
● It is an instrument that splits the monochromatic light into two
or the intensity of radiation.
components: one beam passes through the sample, and the other
through a reference solution or blank.
● The additional beam corrects for variation in light source intensity.
● The absorbance of the sample can be recorded directly as the
electrical output of the sample beam.

Types of Double-beam Spectrophotometer

● Double-beam in space - with 2 photodetectors, for the
sample beam and reference beam.
● Double-beam in time - with 1 photodetector; alternately
passes the monochromatic light through the sample
cuvette and then through the reference cuvette using a
chopper (breaks out radiation beam) or rotating sector
Mirror.
Six Basic Components of Single or Double-beam Spectrophotometer
1. Stable source of radiant energy Diffraction gratings
2. Filter that isolates a specific region of the electromagnetic ● These are the most commonly used; better resolution than prism.
spectrum, ● These are made by cutting grooves (parallel grooves) or slits into an
3. Sample holder aluminized surface of a flat piece of crown glass; wavelengths are bent as they
4. Radiation detector pass a sharp corner.
5. Signal processor ● Holographic gratings are special types of diffraction gratings.
6. Readout device
Filters
Parts of the Spectrophotometer ● These are simple, least expensive, not precise, but useful monochromators.
1. Light/Radiant Source ● These are made by placing semi-transparent silver films on both sides of a
● It provides polychromatic light and must generate sufficient radiant energy dielectric such as magnesium fluoride.
or power to measure the analyte of interest. ● Filters produce monochromatic light based on the principle of constructive
● An intense beam of light is directed through the monochromator and the interference of waves - light waves enter one side of the filter and are
sample. reflected at the second surface.
● To give accurate absorbance measurements throughout its absorbance ● Filters usually pass a wide band of radiant energy and have a low
range, its response to change in light intensity must be linear. transmittance of the selected wavelength.

Types of Light Source 4. Exit Slit

● Continuum source - emits radiation that changes in ● It controls the width of the light beam (bandpass); allows only a
intensity; widely used in the laboratory narrow fraction of the spectrum to reach the sample cuvette.
○ Examples: tungsten, deuterium, and xenon lamps ● Bandpass is the total range of wavelengths transmitted.
○ Tungsten light bulb is the commonly used light source in the visible ● Accurate absorbance measurement requires a bandpass less
and near infrared region. than 1/5 the natural bandpass of the spectrophotometer.
○ Deuterium lamp is routinely used to provide UV radiation in analytic ● Spectral purity of the spectrophotometer is reflected by the
spectrometers. bandpass, that is, the narrower the bandpass, the greater the
○ Xenon discharge lamp produces a continuous source of resolution.
radiation, which covers both the UV and the visible range.
● Line source - emits limited radiation and wavelength 5. Cuvette
○ Examples: Mercury and sodium vapor lamps in spectrophotometers ● It is also called absorption cell/analytical cell/sample cell.
(UV and visible regions), and the hollow cathode lamp (AAS) ● It holds the solution whose concentration is to be measured.
○ Line sources that emit a few discrete lines find wide use in atomic
absorption, molecular, and fluorescent spectroscopy.
○ Light Amplification by Stimulated Emission of Radiation (LASER) is
also used as light sources for spectrophotometry.

Factors for Choosing a Light Source

● Range, spectral distribution within the range, the source of
radiant production, stability of the radiant energy, and Kinds of Cuvettes
temperature. ● Alumina silica glass - most commonly used (available in 350 nm
● Alternatives for Tungsten Light Bulb to 2000 nm)
○ Mercury arc (visible and UV) ● Quartz / plastic - used for measurement of solution requiring visible and
○ Deuterium lamp (165 nm) - UV ultraviolet spectra
○ Hydrogen lamp - UV ● Borosilicate glass
○ Xenon lamp - UV ● Soft glass
○ Merst glower - IR
○ Lobar (silicone carbide) - IR 6. Photodetector
● It detects and converts transmitted light into photoelectric energy.
2. Entrance Slit ● It detects the amount of light that passes through the sample
● It minimizes unwanted or stray light and prevents the entrance of scattered cuvette.
light into the monochromator system.
● Stray light refers to any wavelengths outside the band transmitted by the
monochromator; it does not originate from the polychromatic light source; it
causes absorbance error.
● Stray light limits the maximum absorbance that a spectrophotometer can
achieve.
● Straylight is the most common cause of loss of linearity at high-analyte
concentration.

3. Monochromator Kinds of Photodetectors

● It isolates specific or individual wavelengths of light. Barrier Layer Cell / Photocell / Photovoltaic Cell
● It is the simplest detector; least expensive; temperature-sensitive.
● It is used in filter photometers with a wide bandpass.
● It is the basic photodetector - for detecting and measuring radiation in the
visible region.
● It is composed of selenium on a plate of iron covered with transparent layer
of silver.
● This cell typically has a maximum sensitivity at about 550 nm and the
response falls off to about 10% of the maximum at 350 and 750 mm.

Phototube
Kinds of Monochromators
● It contains cathode and anode enclosed in a glass case.
Prisms
● It has a photosensitive material that gives off electrons when
● These are wedge-shaped pieces of glass, quartz, or sodium chloride.
light energy strikes it.
● These can be rotated, allowing only the desired wavelength to pass
● It requires an external voltage for operation.
through an exit slit.
● A narrow light focused on a prism is refracted as it enters the more
dense glass.
 Photomultiplier Tube (PMT)
● It is the most sensitive photodetector. Absorbance (A)
● The response of the PMT begins when incoming photons strike a ● It is the amount of light absorbed.
photocathode. ● It is proportional to the inverse log of transmittance.
● It is limited to measuring low power radiation because intense light causes ● It is mathematically derived from % T.
irreversible damage to the photoelectric surface.
● It should never be exposed to room light because it will be damaged. A = abc = 2 - log%T
where:
A = Absorbance
a = Molar absorptivity; absorptivity of the compound under standard
conditions
b = Length of light through the solution
c = Concentration of absorbing molecules/solution

Photodiode
● It is not as sensitive as PMT (Photomultiplier Tube) but with excellent
linearity.
● It measures light at a multitude of wavelengths - detects less amount of
light.
● It is most useful as a simultaneous multichannel detector.
Where:
7. Meter or Readout Device Au → Absorbance of the Unknown Solution
● It displays the output of the detection system. As → Absorbance of the Standard Solution
● Example: Galvanometer / ammeter / light-emitting diode (LED) display Cs → Concentration of the Standard Solution

Percent Transmittance (%)

It is the ratio of the radiant energy transmitted (T) divided by the radiant
energy incident on the sample (I).

Quality Assurance Monitoring of Spectrophotometers where:

● It includes monitoring the wavelength/photometric accuracy, absorbance It = Transmitted light thru the sample
check, linearity, and detects straylight. Io= Intensity of light striking the sample
● Wavelength accuracy is the actual wavelength of light that passes through ● The %T measured by commercial spectrophotometers is the ratio of the
the monochromator. sample transmitted beam divided by the blank transmitted beam (sample
● Didymium or holmium oxide filter is used to check wavelength accuracy beam signal/blank beam signal × 100).
(wavelength quality assurance calibration). ● In actual practice, the light transmitted by a blank is substituted for Io.
● Didymium glass has an absorption peak around 600 nm.
● Holmium oxide has multiple absorption with a sharp peak at 360 nm.
● Neutral density filters and dichromate solution verify linearity.
● An absorbance check is performed using glass filters or solutions that have
known absorbance values for a specific wavelength - the operator simply
measures the absorbance of each solution at a specified wavelength and
compares the results with the stated values.
● Absorption spectroscopy is preferred for solutions with absorbance values of
less than 2.0.
● Absorbance values higher than 2.0 may yield unreliable results, and thereby
cause a deviation from Beer's law, resulting in bending of the linear plot.

Beer’s Law
● It states that the concentration of the unknown substance is
directly proportional to the absorbed light (absorbance or optical
density) and inversely proportional to the amount of transmitted
light (% Transmittance).
● It mathematically establishes the relationship between
concentration and absorbance.
 II. VOLUMETRIC (Titrimetric)
● Principle: The unknown sample is made to react with a known solution in
the presence of an indicator.
○ Examples:
■ Schales and Schales method (Chloride test)
■ EDTA Titration method (Calcium test)

III. TURBIDIMETRY
● Principle: It determines the amount of light blocked (reduction of light)
by a particulate matter in a turbid solution.
● It depends on specimen concentration and particle size.
● The measurement of the reduction of light is due to particle formation.
● For measuring abundant large particles (proteins) and bacterial suspensions.
● Solutions requiring quantitation by turbidimetry are measured using visible
photometers or visible spectrophotometers.
● Uses: Protein measurements (CS and urine); to detect bacterial growth in
broth cultures, antimicrobial test (broth method); and to detect clot formation

IV. NEPHELOMETRY
Blanking Technique
● Principle: It determines the amount of scattered light by a
● It means the blank contains serum but without the reagent to complete the
particulate matter suspended in a turbid solution.
assay.
● Photodetector: Photomultiplier tube
● Reagent blank corrects absorbance caused by the color of the reagents - the
● Use: For measuring the amount of antigen-antibody complexes (proteins)
absorbance of reagents is automatically subtracted from each of unknown
● Light scattering depends on wavelength and particle size.
reading.
● Light scattered by particles is measured at an angle, typically 15 to 90
● Sample blank measures absorbance of the sample and reagent in the
degrees to the beam incident on the cuvette.
absence of the end product, and corrects the measurement for optical
● Most antigen-antibody complexes have a diameter of 250 nm to 1500 mm,
interference (like hemoglobin) absorbing the wavelength of measurement.
and the wavelengths used are 320 mm to 550 nm, thus light is scattered
● A blanking process may not be effective in some cases of turbidity, and
forward (Rayleigh-Debye type).
ultracentrifugation may be necessary to clear the serum or plasma of
● Basic Components: Light source (mercury-arc lamp, a tungsten-filament
chylomicrons.
lamp, light emitting diode, and a laser), collimator, monochromator, sample
● Lipids interfere mainly by increasing light scatter (turbidity).
cuvette, stray light trap, and photodetector.
● To correct for artifactual absorbance readings, "blanking"
procedures or dual-wavelength methods may be used.

B. Flame Emission Photometry (FEP)

● Principle: Excitation of electrons from lower to higher energy state.
● Light source: Flame (also serves as the cuvette)
● Method: Indirect Internal Standard Method
● Internal standard: Lithium/Cesium - corrects variations in flame and
atomizer characteristics
● It measures the light emitted by a single atom burned in a flame.
● It is used in the measurement of excited ions (sodium and potassium).
● Flickering light indicates changes in the fuel reading of the instrument.

C. Atomic Absorption Spectrophotometry (AAS)

● Principle: Element is not excited but merely dissociated from its chemical
bond and place in an unionized, unexcited, ground state.
● Light source: Hollow-cathode lamp
V. FLUOROMETRY
● Interferences: Chemical, matrix (differences in viscosity), and ionization ● Principle: It determines the amount of light emitted by a molecule after
● It measures the light absorbed by atoms dissociated by heat. excitation by electromagnetic radiation.
● It is used for measurement of unexcited trace metals (calcium and ● It measures the amount of light intensity present over a zero background.
magnesium). ● It uses two monochromators (either filters, prisms, or gratings) the
● It is more sensitive than FEP; it is accurate, precise, and very specific. wavelength that is best absorbed by the solution to be measured is selected
● Internal standard is not needed - changes in aspiration have little effect on by the primary filter or excitation monochromator; the incident light is
the number of ground state atoms. prevented from striking the photodetector by the secondary filter or emission
● An atomizer (nebulizer/graphite furnace) is used to convert ions to atoms; a monochromator.
chopper is used to modulate the light source. ● To avoid potential interference from the excitation signal,
● Lanthanum or strontium chloride is added to samples to form stable fluorescence measurements detect the emitted light at a right
complexes with phosphate. angle to the incident light.
● It is 1000x more sensitive than spectrophotometer - emitted
radiation is measured directly.
● It is affected by quenching - pH and temperature changes,
chemical contaminants, and UV light changes.
● Fluorescence is the light emission from a single excited state,
while phosphorescence is the light emission from an excited
triplet state.
● Use: For measurement of porphyrins, magnesium, calcium, and
catecholamines
● Basic components: Light source (mercury arc or xenon lamp), primary
monochromator, secondary monochromator, cuvette, and photodetector
(photomultiplier tube or phototube).
 Vycor (Corning)
● It is utilized for high thermal, drastic heat shock, and extreme chemical
treatment with acids (except hydrofluoric) and dilute alkali; it can be heated to
900°C.

Flint Glass (Soda lime)

● It is made up of soda-lime glass and a mixture of calcium, silicon, and
sodium oxides.
● It is used to make disposable glassware.
● It has poor thermal resistance; easy to melt.

VI. CHEMILUMINESCENCE
● Principle: The chemical reaction yields an electronically excited compound
that emits light as it returns to its ground state or transfers its energy to
another compound which then produces an emission.
● It quantifies sample analytes in a wide range of wavelengths not observed in
the visible spectrum.
● This method is best when differentiating two compounds having excitation Cleaning of Glassware
reaction at the same wavelength but emit at different wavelengths. ● Pre Soaking glassware in soapy water.
● It is widely used nowadays due to its high sensitivity while even, more ● Multiple rinses must be done with appropriate grade water.
sensitive than fluorometry. ● Detergent-contaminated water have more alkaline ph as compared with the
● Methods: Chemiluminescence Immunoassay (CLIA) ph of the appropriate grade water.
● Uses: Autoantibody testing; measurement of hormones, drugs, vitamins, ● Acid dichromate and nitric acid is the cleaning solution for glassware.
tumor markers; forensic analysis; microbial and infectious disease marker ● Ultrasonic cleaners help remove debris coating the surfaces of glass or
studies; and toxicology Photodetector: Photomultiplier tube (luminometer). plasticware.

INSTRUMENTATION AND AUTOMATION PIPETTE CLASSIFICATION

TYPES OF GLASSWARE I. CALIBRATION MARKS/DESIGN

Borosilicate Glass (Pyrex and Kimax) To Deliver (TD) Pipette
● It is used for heating and sterilization purposes; most commonly used. ● It delivers the exact amount it holds into a container; should not be
● It is characterized by a high degree of thermal resistance, has low alkali blown out.
content, and is free from the magnesium lime-zinc group of elements. ● It is designed to meet the requirements of Class A-type pipettes.
● It does not contain heavy metals, arsenic, and antimony. ● It is designed to drain by gravity, and the desired volume is obtained when
● Examples: Pyrex and Kimax glassware draining stops.
● Strain point: 515° C (Pyrex) ● Proper usage: Hold vertically with the tip against the side of the container
and should not touch the accumulating liquid.
● Examples: Serologic, Mohr, and volumetric transfer pipettes

I. CALIBRATION MARKS/DESIGN
To Contain (TC) Pipette
● It holds a particular volume but does not dispense the exact volume.
● It is also known as the rinse-out pipette.
Boron-free Glassware/Soft Glass
● It does not meet Class A certification criteria.
● It has high resistance to alkali.
● Examples: Sahli hemoglobin and Long-Levy pipettes
● Its thermal resistance is less as compared to borosilicate glass.
II. DRAINAGE CHARACTERISTICS
Blow-out Pipette
● It has continuous etched rings on top or near the mouthpiece of the pipette
and exact volume is obtained when the last drop is blown out.
● This type of pipette is not rinsed out.
● Examples: Ostwald-Folin and serological pipette
● Self-draining Pipette
● It allows the liquid to drain by gravity.
● It does not have etched or frosted rings.
Corex
● It is a special alumina-silicate glass that has been strengthened chemically III. TYPES OF PIPETTES
than thermally; six times stronger than borosilicate. Transfer Pipette
● Volumetric Pipette - for nonviscous fluid; self-draining; small amount left in
the tip should not be blown out
● Ostwald Folin - for viscous fluid; with etched ring
● Pasteur Pipette - transfers fluids without consideration of a specific volume
● Automatic macro or micropipettes

Graduated or Measuring Pipette

● Serological Pipette - with graduations to the tip; blow-out pipette
● Mohr Pipette - without graduations to the tip; calibrated between 2 marks;
self-draining Pipette
● Bacteriologic Pipette
● Ball, Kolmer, and Kahn Pipette
● TC pipettes - Glass Micropipettes (< 1 mL)
○ Sahli-Hellige Pipette
○ Lang-Levy Pipette
○ RBC and WBC Pipettes
○ Kirk and Overflow Pipettes

Dispenser / Dilutor Pipette

● It obtains liquid from a common reservoir and dispenses it repeatedly.
● It combines sampling and dispensing functions.

MECHANICAL/AUTOMATIC PIPETTES
Air Displacement Pipette
● It relies on piston for suction creation to draw the sample into a disposable
tip.
● The piston does not come in contact with the liquid.
● A disposable, one-time use polypropylene tip is attached to the pipette
barrel.

For Calibration of Pipettes and Glassware

● Class A pipettes do not require recalibration. To prepare a piece ofglassware
for calibration, thoroughly wash and dry it using appropriate cleaning
procedures.
● Distilled water is the calibrating medium for TD pipettes, while mercury is for
Positive Displacement Pipette TC pipettes.
● It operates by moving the piston in the pipette tip or barrel, much like a ● Gravimetric and spectrometric methods verify the accuracy and precision of
hypodermic syringe. pipette volumes.
● It does not require a different tip for each use. ● 0.1% phenol red solution in distilled water is utilized to compare the
● It utilizes a Teflon-tipped plunger that fits tightly inside the capillary which is reproducibility of brands of pipette tips.
either a siliconized glass or plastic. ● Micropipettes must be checked for accuracy and precision before they are
● It is useful if a reagent reacts to plastics. used.
● Volumetric flasks for the preparation of standards and other solutions must
meet Class A specifications as defined by the National Institute of Standards
and Technology (NIST).
● Any glassware that fails to fulfill the Class A tolerance should be
discarded.

Maintenance Check of Automatic Pipettes

● Air displacement pipette has a fixed stroke length that requires regular
preventive maintenance monitoring.
● Air displacement pipette has a seal to prevent air leakage into the pipette
when the piston is moved, and these seals require periodic greasing to
maintain their integrity.
● Positive displacement pipettes need to have their spring checked and the
Teflon tip replaced periodically.
● A slide wire is used to quickly check the plunger setting of positive
displacement pipettes.
Volume Measurements
1 lambda = 1 microliter (0.001 mL)
1 microliter = 1.0 milligram
Examples of plastic materials for laboratory use
1. Polycarbonate - tubes for centrifugation, graduated cylinders, and
flasks
2. Polyethylene - test tubes, bottles, stoppers, disposable transfer
pipettes, volumetric flasks, and graduated tubes
3. Teflon - stirring bars, tubing, cryogenic vials, and bottle cap liners;
almost chemically inert; suitable for extreme temperature treatment (-70 °C to
200°C); resistant to strong chemicals (acids, bases, alcohol, and hydrocarbons) Centrifugal Analyzer
● It uses the force generated by centrifugation to transfer specimen and
Calibration of Analytical Balance and Thermometer reagents.
Analytical Balance ● Liquids are placed in separate cuvettes for measurement at the perimeter of
● Laboratory balances require calibration at regular intervals - calibration a spinning rotor (1000 rpm).
intervals should coincide with the requirements of the laboratory's ● It uses acceleration and deceleration of the rotor to transfer the reagents
licensing and accrediting organizations. and sample from one chamber to another.
● The operator must avoid direct contact with the weights by using clean ● It is also known as a "run-based" system which means it cannot be
gloves or special lifting tools (e.g., forceps). Hand contact with the weights can interrupted once the batch testing/ measurement has started.
cause corrosion. ● Mixing: Centrifugal force (rotor) is utilized for bubbling of air
● Examples: Cobas-Bio (Roche) and IL Monarch
● Major advantage: Batch analysis (discrete-batch type system)
● Requirement: Each cuvette should be uniformly matched with each other to
achieve reliable result.

Thermometer
● Temperature-monitoring devices should be verified for accuracy at 6or
12-month intervals.
● Types of thermometers: Total immersion (freezers and refrigerators)
and Partial immersion (water baths and heating blocks)

THREE BASIC APPROACHES TO AUTOMATION

Advantages
1. Increases the number of tests to be performed in a given period Discrete Analyzer
2. Minimizes variation of result from one laboratorian to another ● It is the most popular and versatile analyzer - measures only
3. Eliminates the potential error in manual analyses such as the tests requested on a sample.
pipetting, calculation, and transcription of results ● It requires 2 - 6 uL of the sample (minimum volume).
● It employs a variety of syringe pipettes (positive
Steps in Automated Analysis liquid-displacement pipettes) to aspirate and dispense samples
1. Specimen Preparation and Identification and reagents, and with a single analytic pathway.
2. Specimen Measurement and Delivery ● It is capable of running multiple tests one sample at a time.
3. Reagent System and Delivery ● Each sample-reagent mixture is handled separately in its own reaction
4. Chemical Reaction Phase (samples and reagents are mixed together) vessel, and throughout is based on the number of tests run per hour, not the
5. Separation and Incubation number of samples per hour.
6. Measurement Phase ● Random access analyzers choose samples and reagents at random
7. Signal Processing and Data Handling regardless of their placement in the automated platform.
8. LIS Support ● For dry slide technology (reflectance photometry), the spreading layer
permits a rapid uniform spreading layer over the reagent layer.
Types of Automation ● Mixing: Magnetic driven teflon stirring bar, forceful dispensing, magnetic
Continuous Flow Analyzer stirring bars, rotating paddle, and ultrasonic energy
● Liquids are pumped through a system of continuous tubing. ● Examples: Vitros, Dimension Dade, Beckman ASTRA System, Hitachi, Bayer
● Samples flow through a common reaction vessel or pathway. Advia, Roche Cobas Integra, and Analytics P Module
● Analysis of samples is through sequential testing in a single or multiple ● Major advantage: Random access capability - allows STAl samples to be
parallel channel configuration, which means the same group of tests is easily tested
performed on each sample regardless of which tests were ordered.
● Air bubbles at regular intervals serve as separating and cleaning media.
● A heating bath maintains the required temperature of the reaction to
allow complete color development (same with discrete analyzers)reaction rate
is controlled by temperature.
● It can also perform batch analysis (Bishop et al., 2018).
● Mixing of sample and reagents: By using a glass coil inserted into the flow
path
● Advantage: Assists the laboratory that needs to process several samples but
same test/procedure
Sampling and Reagent System
● Disadvantage: Carry-over contamination and "continuous" waste of reagents
● Barcode-labeled tubes with patient information and test requests are used
● Examples: Simultaneous Multiple Analyzer (SMA) and Technicon
as specimen holders in automation.
● Rectangular racks or carousels with corresponding number codes are utilized
as sample containers/stations in the loading zone of the modular platforms.
● Liquid level sensors detect the presence of inadequate sample as they scan
or travel into the sample container.
● Most analyzers will automatically reconstitute a lyophilized reagent.
Types of Reagent System
1. Open reagent system - Reagents other than manufacturer's
reagents can be utilized for measurement.
2. Closed reagent system - The operator can only use the
manufacturer's reagents.

Sources of Errors in Automation

● Carry over contamination
● Sample evaporation
● Presence of clot
● Inadequate sample
● Reusable probes

Steps to Reduce Carryover Contamination

● Aspirate a wash solution between pipetting cycles;
back-flush; use disposable plastic pipette tips to transfer
each sample; and clean the probe using a wash solution.

Examples of Detection System in Automation

● Absorption or reflectance spectroscopy; ion-selective
electrodes; and some immunoassays that may be based on
chemiluminescence

Example of a Stand-Alone-Automated Platform

● The Automate 800-Beckman Coulter is an all-in-one
biochemistry analyzer which can perform the following
procedures aside from the measurement and LIS phases:
sample receipt, sorting, centrifugation, decapping, sample
volume detection, aliquoting, and sample storage.

Radio-Frequency Identification (RFID)

● It is a system for tracking samples (tagged samples), starting from the
collection of specimens down to release of laboratory results.
● The demographics of the patient are encrypted in the "chip" including the
laboratory tests requested for that individual.
● It also utilized for managing reagent supplies and storage.

Reflectance Photometry
● It is the measurement of light reflected from solid surfaces.
● The intensity of the reflected light from the reagent carrier is
compared with the intensity of light reflected from a
reference surface.
● A reflectometer is used to measure analytes by measuring
the quantity of light reflected by a liquid sample that has
been dispensed onto a grainy or fibrous solid support.