INSTRUMENTATION
Instrumentation
TYPES OF GLASWARE
1. Borosilicate glass (pyrex and kimax)
• It is used to heating and sterilization purposes, this is the
most commomly used glass.
• It is characterized by a high degree of thermal resistance, has
low alkali content, and is free from the magnesium-zinc group
of elements, heavy metals, arsenic, and antimony.
• Strain point: 515° C ( Pyrex)
Instrumentation
TYPES OF GLASWARE
2. Boron-free glassware/ Soft glass
• It has high resistance to alkali
• Its thermal resistance is less as compared to borosilicate
glass.
3. Corex (Corning)
• Is a special alumina-silicate glass that has been
strengthened chemically than thermally; six times stronger
than borosilicate
Instrumentation
TYPES OF GLASWARE
4. Vycor (Corning)
• Its 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
5. Flint Glass
• It has made up of soda-lime glass and a mixture of
calcium, silicon, and sodium oxides.
• It has poor resistance to high temperatures - easy to melt
and used to make disposable glassware.
Instrumentation
PIPET CLASSIFICATION
I. CALIBRATION MARKS/DESIGN
1. To Deliver (TD) – It delivers the excat amount it holds into a
container
2. To Contain (TC) – It holds the particular volume but does not
dispense the exact volume.
II. DRAINAGE CHARACTERISTICS
1. Blowout – It has a continuous etched rings on top of the
pipet; exact amount is obtained when the last drop is blown
out.
2. Absence of etched rings; liquid is allowed to drain by gravity.
Instrumentation
PIPET CLASSIFICATION
III. TYPES
1. Transfer Pipet
• Volumetric Pipet – for non-viscous fluid; self draining;
small amount left in the tip should not be blown out.
• Ostwald Pipet – for viscous fluid; with etched ring
• Pasteur Pipet – transfers fluids without consideration of
a specific volume
• Automatic macro – or micropipettes
Instrumentation
PIPET CLASSIFICATION
III. TYPES
2. Graduated or Measuring Pipet
• Serological Pipet – with graduations to the tip; blowout
pipet
• Mohr Pipet – without graduations to the tip; calibrated
between 2 marks; self- draining pipet
• Bacteriologic Pipet
• Ball, Kolmer and Kahn Pipet
• Micropipets (<1ml) – TC Pipets
1. Sahli-Hellige pipet
2. Lang-Levy pipets
3. RBC and WBC pipets
4. Kirk and Overflow Pipet
Instrumentation
Mechanical or Automatic Pipets
a. Air Displacement Pipet
• 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.
b. Positive Displacement Pipet
• It operates by moving the piston in the pipet tip or barrel,
much like a hypodermic syringe.
• It does not require a different tip for each use.
c. Dispenser/Dilutor Pipet
• It obtains liquid from a common reservoir and dispensed it
repeatedly.
• It combines sampling and dispensing functions.
Instrumentation
Mechanical or Automatic Pipets
For Calibration of Pipettes
§ Class A Pipettes do not require recalibration
§ Distilled water is the calibrating medium for TD pipettes
while mercury is for TC Pipettes.
§ Gravimetric and spectrometric methods – used to verify
pipette volume accuracy and precision
§ 0.1% phenol red solution in distilled water – used to
compare the reproductivity of brands of pipette tips
Volume Measurements
• 1 lambda = 1 microliter ( 0.001 mL)
• 1 microliter = 1.0 milligram
Instrumentation
NOTES TO REMEMBER
Ø Plastic pipet tips are made primarily of polypropylene
Ø As the fluid is allowed to drain into the receiving vessel,
pipets should be held in a vertical position with the tips
against the side of the receptacle.
Ø For volumetric TD pipettes, it should not be shaken or hit
against the wall of the container during draining because
any disruption of the free-flowing liquid may result in an
inaccurate deliver of the liquid.
Ø Imperfect wetting or the presence of discreet droplets of
water indicates that the pipette is not sufficiently clean.
Ø Acid dichromate is a cleaning solution for glassware.
Instrumentation
CALIBRATION OF ANALYTICAL BALANCE AND THERMOMETER
Analytical Balance
§ Laboratory balances require calibration at regular intervals –
calibration intervals should coincide with the requirements of the
laboratory’s licensing and accrediting organizations.
§ The new mass standards and test weight accuracy classes
appropriate for laboratory balances include the American Society for
Testing Materials (ASTM) classes 1 and 2.
§ The operator must avoid direct contact with the weights by using
clean gloves or special lifting tools (e.g., forceps). Hand contact with
the weights can cause corrosion.
Instrumentation
CALIBRATION OF ANALYTICAL BALANCE AND THERMOMETER
Analytical Balance
THERMOMETER
v 2 types of thermometers: total immersion (freezers and refrigerators)
partial immersion (water baths and heating blocks)
Instrumentation
CALIBRATION OF ANALYTICAL BALANCE AND THERMOMETER
Analytical Balance
THERMOMETER
v Noncertified thermometers can be calibrated by using an NIST SRM934
thermometer or an NIST SRM 1968 gallium melting point cell.
NIST SRM934
v Temperature-monitoring devices should be verified for accuracy at 6 or
12 months intervals.
Instrumentation
THREE BASIC APPROACHES TO AUTOMATION
Advantages:
1. Increase the number of tests to be performed in a
given period.
2. Minimizes variation of result from one laboratorian to
another.
3. Eliminates the potential error in manual analyses such
as pipetting, calculation and transcription of results.
Instrumentation
1. CONTINUOUS FLOW ANALYZER
§ Liquids are pumped through a system of continuous tubing.
§ Samples flow through a common reaction vessel or pathway.
§ Air bubbles at regular intervals serve 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.
§ Mixing of sample and reagents: by using a glass coil inserted into the flow path
§ Example: Simultaneous Multiple Analyzer (SMA), Technicon
§ Disadvantage: all tests are performed in parallel (measurement of every analyte
configured on the system for every sample)
Instrumentation
2. CENTRIFUGAL ANALYZER
§ It uses the force generated by centrifugation to transfer specimen and reagents.
§ Liquids are placed in separate cuvets for measurement at the perimeter of a spinning
rotor (1000rpm).
§ It uses acceleration and deceleration of the rotor to transfer the reagents and sample
from one chamber to another.
§ Mixing: Centrifugal force (rotor) is utilized or bubbling of air
§ Examples: Cobas-Bio (Roche) and IL Monarch
§ Major Advantage: Batch analysis ( discrete-batch type system)
Cobas-Bio (Roche)
Instrumentation
3. DISCRETE ANALYZER
§ It is the most popular and versatile analyzer – measures only the tests
requested on a sample.
§ It requires 2-6µL of the sample (minimum volume).
§ It employs a variety or syringe pipettes (positive liquid-displacement
pipets) to aspirate and dispense samples and reagents.
§ It is capable of running multiple-test-one-sample-at-a-time.
§ Each sample-reagent mixture handled separately in its own reaction
vessel.
§ For dry slide technology (reflectance photometry), the spreading layer
permits a rapid uniform spreading layer over the reagent layer.
§ Mixing: magnetic driven teflon stirring bar, forceful dispensing,
magnetic stirring bars, rotating paddle, and ultrasonic energy.
§ Examples: Vitros, Dimension Dade, Beckman ASTRA System, Hitachi
Bayer Advia, Roche Cobas, Integra and Analytics P Module
§ Major advantage: Random access capability – allows STAT samples to
be easily tested.
Instrumentation
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 the 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.
Instrumentation
Reflectance photometry
NOTES TO REMEMBER
§ For measurement, visible and UV light
spectrophotometry are the common methods.
§ Abbot TDX uses fluorescent polarization for drug
analysis.
§ Tablet form reagents – ACA Star (Dade) and
Paramax analyzers.
§ Multilayered dry slide (thin film) reagents – Vitros
analyzer
§ Carry over is the transport of quantity of analyte or
reagent from one specimen reaction into another,
and contaminating a subsequent one.
Instrumentation
Terminologies:
1. Batch testing – all samples are loaded at the same time, and a
single test is conducted on each sample.
2. Parallel testing – more the one test is analyzed concurrently on a
given clinical specimen.
3. Random access testing – any test can be performed on any sample
in any sequence.
4. Sequential testing – multiple tests analyzed one after another on a
given specimen.
5. Open reagent system – a system other than manufacturer’s
reagents can be utilized for measurement
6. Closed reagent system – a system where the operator can only be
manufacturer’s reagents
7. Pneumatic tube delivery system – it provides point-to-point delivery
specimens to the laboratory and offered several advantages over
specimen transport by humans.
