Chapter- Two

2. LABORATORY WARES AND EQUIPMENTS

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Learning objectives
At the end of this chapter, the student will be able to:
1. State the different laboratory wares.
2. Describe the use of laboratory wares.
3. Explain the general cleaning and care of laboratory wares.
4. Identify the types and uses of laboratory balances.
5. Explain the advantages of laboratory refrigerators.
6. Describe the importance of ovens, water baths and
incubators.
7. State the use of photometers and desiccators.
8. Identify the types and uses of microscopes.
9. State the basic components centrifuge
10. Discuss pH meter in terms of ion activity and units.
11. Describe the main components of a pH meter including
3
their role in analysis.
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Outline
2. LABORATORY EQUIPMENTS AND WARES
2.1: General laboratory wares
2.2 lab equipments
2.2.1: Microscope
2.2. 2: Equipment for purifying water
2.2.3: Equipment for weighing
2.2.4: Equipment for pipetting and dispensing
2.2.5: Laboratory centrifuges
2.2.6: laboratory autoclaves, ovens
2.2.7: Incubator, water bath, heat block
2.2.8: Colorimeter
2.2. 9: Mixers
2.2.10: Refrigerators
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Outline…
2.2.11: Desiccators
2.2.12: PH meter
2.2.13: Safety cabinets
2.3: Care and cleaning of laboratory equipments and wares

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2.1: General laboratory wares
LABORATORY GLASSWARES AND PLASTICWARES
Definition: laboratory glassware and plastic wares are materials
used in clinical lab. For:
 measuring

 pipetting

 transferring

 Preparation of reagents

 Storage etc.

 Most of the routine laboratory wares used to be of glass, but

recent advantage made in the use of plastic resin to
manufacture a wide range of plastic ware having led to a
gradual replacement of glass wares with durable plastic ware.

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2.1.1 Classification of Laboratory glass wares

A. can be divided in to five main types according to their

composion
1. Glass with high thermal resistance – borosilicate glass can
resist about 500oc and low alkaline contact.
2. High silica glass- contains 96% silicon, chemically stable and
electric resistant. It is strengthened chemically rather than
thermally.
3. Glass with high resistance to alkali- Boron free, used in strong
alkali low thermal resistance.
4. Low actinic glass – amber color to protect light
5. Standard flint glass- soda lime glass, poor resistance to
increased temp. Contains free soda in its walls.
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Classification of Laboratory glass……
B . Based on their use
a) volumetric wares
b) Semi-volumetric Glass wares
c) Non- volumetric glass wares.

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Classification of Laboratory glass……
a)Volumetric wares
 Apparatus used for measurement of liquids
 Can be made either from glass or plastic . it includes :
 Volumetric flasks

 Graduated centrifuge tubes

 Graduated serological pipette

 Medicine dropper

 Burettes

 Micropipettes

 Diluting or thoma pipettes etc

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Classification of Laboratory glass……
b). Non- volumetric glass wares
 Erlenmeyer flask

 Round bottom flask

 Flat bottom flask

 Beaker

 Centrifuge tube

 Test tube

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Classification of Laboratory glass……
C ).Semi-volumetric Glass wares: are used for approximate
measurement. it includes;
 These glass wares includes

 Graduated cylinder

 Graduated specimen glass

 Beakers

 Conical flask

 Medicine droppers with or with out calibration mark

 Graduated beaker with double beaks

 Graduated glass

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05/15/24 F I G U R E : Laboratory glassware 12
2.1.2 Pipettes
 There are several types each having their own advantages and
limitations.
 They are designated as class “A” or “B” according to their
accuracy.
 Class “A” pipettes are the most accurate and the
tolerance limits are well defined that is, +0.01, + 0.02 and
0.04 ml for 5, 25, and 50 ml pipettes respectively.
 Class “B” pipettes: are less accurate but quite satisfactory
for most general laboratory purposes.
 Significant errors will result if the temperature of the liquid
pipetted is widely different from the temperature of calibration.
 The usual temperature of calibration is 20oC and this is marked
on the pipette.
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2.1.2.1 Volumetric pipettes
 Volumetric pipettes are calibrated to deliver a constant
volume of liquid.
 The most commonly used sizes are 1, 5, and 10ml
capacities.
 Less frequently used sizes are those which deliver 6, 8, 12,
and so on ml.
 They have a bulb mid – way between the mouthpiece and
the tip.
 The Volume (capacity) and calibration temperature of the
pipettes are clearly written on the bulb.

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Volumetric pipettes……
 They should be used when a high degree of accuracy is
desired.
 The pipette is first rinsed several times with a little of the
solution to be used, and then filled to just above the mark.
 Then the liquid is allowed to fall to the mark and the tip is
carefully wiped with filter paper.
 The contents are allowed to drain in to the appropriate
vessel. A certain amount of liquid will remain at the tip and
this must not be blown out.

N.B: The reliability of the calibration of the volumetric pipette decreases with an
increase in size and therefore, special micropipettes have been developing
for chemical microanalysis.

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2.1.2.2 Graduated or measuring pipettes
 Graduated pipettes consist of a glass tube of uniform bore with
marks evenly spaced along the length.
 The interval between the calibration marks depends up on the
size of the pipette.
Two types calibration for delivery are available.
 These are:
A. One is calibrated between two marks on the stem (Mohr).
B. The other has graduation marks down to the tip (serological
pipette)

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Graduated or measuring…….
 These pipettes are intended for the delivery of
predetermined volumes.
 The serological pipette must be blown out to deliver the
entire Volume of the liquid and it has an etched ring (pair of
rings) near the mouth end of the pipette signifying that it is a
blow out pipette.
 Measuring pipettes are common only in 0.1, 0.2, 0.5, 1.0
5.0, and 10.0 ml sizes.
 The liquid is delivered by allowing it to fall from one
calibration mark to another.
N.B. The classification of pipettes may not always be based on
the presence or absence of a bulb and etched ring.

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 A. B C D.

A. Volumetric (transfer) B. Ostwald folin (transfer). C. Measuring (Mohr) D. Serological (Graduated)

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Con't…

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 2.1.2.3 Micropipettes
 Micropipettes are frequently used in
 Medical chemistry
 Virology
 Immunology and serology laboratories.
 This is because in these laboratories often only small quantities
of materials are available for measurement.
 They are found in different capacities such as 5, 10, 25, 50, 100
and 1000 micro liter.
 There are also other kinds of pipettes that are used in medical
laboratories
Example: Toma pipette, Pasteur pipette, automatic pipettes
and others

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Automatic
Micropipettes

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2.1.3 Burettes
• Burettes are used for measuring variable quantities of liquid
that are used in volumetric titrations.
• They are made in capacities from 1 to100 milliliters.
• They are long graduated tubes of uniform bore and are
closed at the lower end by means of a glass stopper, which
should be lightly greased for smooth rotation.

Fig. Burette

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2.1.4 Flasks
 There are four types of flaks having 25 to 6,000 milliliter (ml)
capacities.
2.1.4.1 Conical (Erlenmeyer) flasks
 Conical (Erlenmeyer) flasks are useful for titrations and

also for boiling solutions when it is necessary to keep

evaporation to a minimum.
 Some have a side arm suitable for attachment to a

vacuum pump.
2.1.4.2 Flat bottomed round flasks
 Flat-bottomed round flasks are convenient containers to
heat liquids.
 These flasks are widely used in the preparation of
bacteriological culture media.
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2.1.4.3 Round bottomed flasks
 Round bottomed flasks can with stand higher temperatures
than the flat- bottomed type.
 they may be heated in a necked flame or in an electro-
thermal mantle. As a result used for boiling.
2.1.4.4 Volumetric flasks
 Volumetric flasks are
 flat - bottomed
 pear-shaped vessels with long narrow necks
 fitted with ground glass stoppers.

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Volume metric ….
 Most flasks are graduated to contain a certain volume, and
these are marked with the letter’s.
 A horizontal line etched round the neck denotes the stated
volume of water at given temperature.
 They are used to prepare various kinds of solutions.
 The neck is narrow so that slight errors in reading the
meniscus results in relatively small volumetric differences
(minimizes volumetric differences or errors).

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A. Conical B. Flat bottomed C. Round bottomed D.Volumetric

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2.1.5 Beakers
 Beakers have capacities from 5 to 5,000 ml.
 They are usually made up of heat resistant glass and are
available in different shapes.
 The most commonly used is the squat form, which is
cylindrical and has a spout.
 There is also a tall form, usually with out a spout

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2.1.6 Cylinders
 Cylindersare supplied in 10 to 2,000 ml capacities.
 Some are of heat resistant glass or plastic.
 Measurement of liquids can be made quickly with these
vessels, but a high degree of accuracy is impossible
because of the wide bore of the cylinders

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2.1.7 Test tube
 Test tubes are made of hardened glass or plastic
materials that can withstand actions of chemicals,
thermal shock and centrifugal strains.
 They are used to hold samples and solutions during

medical laboratory procedures.

 These include simple round hollow tubes conical

centrifuge tubes, vaccutainer tubes. Test tubes can be

with or with out rims (lips).
 Test tubes with out rim are satisfactory

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2.1.8 Reagent bottles
 Reagent bottles are used to store different types of
laboratory reagents.
 They are made from glass or plastics. Depending on their
use, they are available in various sizes and type.

Dropping bottle 30
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2.1.9 Petri dishes
 Petridishes are flat glass or plastic containers, which have
a number of uses in the medical laboratory.
 They are used predominantly for the cultivation of
organisms on solid media.
 They are made with diameters of 5 to 14 centimeter.

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2.1.10 Funnels
 There are two types of funnels that are widely used in a
medical laboratory. These are filter funnel and separating
funnel.
2.1.10.1 Filter Funnels
 Filter funnels are used for pouring liquids into narrow
mouthed containers, and for supporting filter papers during
filtration.
 They can be made from glass or plastic materials.

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2.1.10.2 Separating funnels
 They are used for separating immiscible liquids of different
densities.
 Separating funnels are used for separating immiscible
liquids of different densities. Example, ether and water.

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2.1.11 Pestle and mortar
 Pestle and mortar are used for grinding solids, for example,
calculi and large crystals of chemicals.
 After each use always clean the pestle and mortar
thoroughly.
 This is because chemicals may be driven into the unglazed
surfaces during grinding, resulting in contamination when
the apparatus is next used

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2.1.12 Laboratory Cuvettes (Photometry)
 used for photometric readings in instruments or used for
measurements of absorbance.
 Glass Cuvettes resist many laboratory reagents like organic
solvents, whereas plastic Cuvettes are affected by many
reagents and become cloudy, hence affecting the
absorbance’ of the reacting mixture and so lack accuracy &
precision.
 Can be glass, quartz, or plastic
 Require uniform thickness, density, composition
 Should be uniformly calibrated

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2. 2 Medical laboratory Equipments
2.2.1: THE MICROSCOPE
 Used to visualize minute objects (animate and inanimate),
that cannot be seen by our naked eye. It is a magnifying
lense.
 It was invented by Anton van Leeuwenhoek –founder of
microscope.
2.2.1.1 Types of microscope
1. Light field microscope
- the group of microscope use light
- It includes:
a. Compound light(bright) field:
 Compound microscope is a light microscope, which is
routinely used in medical laboratories of hospitals and/or36
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b. Dark field microscope or dark ground illumination
 Makes some living micro-organisms which can not be seen
by ordinary transmitted lighting.
Principle
 The light enters a special condenser which has a central
blacked-out area so that the light cannot pass directly to
enter the objective.
 The only light entering the eye comes from the micro-
organisms themselves, no light entering the eye directly from
light source.
 In the way small micro-organisms are seen brightly
illuminated against a black background, like stars in a night
sky.

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Importance of Dark field microscope
 Used for examining-
 Treponema palladium
 Borreliae in blood
 Microfilariae in blood

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c) Phase contrast microscope
 Makes use of this ability of waves to help or hinder each
other to produce variations increase the contrast achieved
by placing annulus in condenser and phase plate in the
objective.
 Used for examination of
 Unstained bacteria
 Urine sediments
 Haemoparasites
 Amoebae in faecal preparations
 Trypanosomes in blood, cerebrospinal fluid, lymph gland fluid

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d) Fluorescence microscope
 widely used in the immunodiagnosis
Principle:
 Ultraviolet light may be used to illuminate particles or micro-
organisms which have been previously stained with
fluorescing dyes.
 These dyes transform the invisible ultraviolet light to visible
light.
 Value of fluorescence microscope
 Examination of sputum and c.s.f for acid fast bacilli (AFB) using an
auramine staining tech.
 Examination of acridine orange stained Trichomonas virginals
flagellates

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2. Electron Microscope: - as the name suggests, employ a
beam of electrons produced by an electron gun to produce the
magnified image.
Mainly used in
 Negative staining
 sample stained with potassium phosphotungestate
 Examination of viruses
NB. The beam can not pass through the metallic back ground of
the microscope

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2.2.1.2 Major parts of microscope
A. Frame work of the microscope
This includes:
 An arm (stand): - The basic frame of the microscope to
which the base, body and stage are attached.
 A stage: - the table of the microscope where the slide or
specimen is placed.
 A foot or base: - is the rectangular part up on which the
whole instruments rest.
B. Focusing system
 This encompasses

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 Coarse and fine focusing adjustments
 Course adjustment: - The course focusing adjustment is
controlled by a pair of large knobs positioned one on each
side of the body. Give rough image.
 Fine adjustment: - it moves the stage so slowly that and
give clear image
 Condenser adjustments: - The condenser is focused usually

by rotating a knob to one side of

it.
 This Moves the condenser up or down.
 The condenser aperture is adjusted by the iris
diaphragm, which is found just below the condenser.
 The principal purpose of the condenser is to
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condense the light required for visualization.
C. Magnification system
This comprises:
 Objectives: - Objectives are components that magnify the
image of the specimen to form the primary image.
 For most routine laboratory work 10x, 40x and 100x (oil immersion)
objectives are adequate.
 Eyepiece:- Eyepiece is the upper optical component that
further magnifies the primary image and brings the light rays
to a focus at the eye point.
 It consists of two lenses mounted at the correct distance.

 It is available in a range of magnifications usually of 10x,

15x and sometimes as high as 20x.

N.B: Based on their number of eyepiece microscopes can be
classified as monocular, binocular microscopes etc.
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D. Illumination system
 Condenser and iris
 Condenser is a large lens with an iris diaphragm.

 The condenser lens receives a beam from the light

source and passes it into the objective.

 The iris is a mechanical device mounted underneath the

Condenser and controls the amount of light entering the

condenser.
 Mirror
 Mirror is situated below the condenser and iris.

 It reflects the beam of light from the light source up wards

through the iris into the condenser.

 The mirror is used to reflect ray or electrical light

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  Many microscopes are now provided with correctly aligned
built-in sources of illumination, which use tungsten or quartz
halogen lamps operating on 6,8 or 12 volts through
variable transforms.
 Filters
 Light filters are used in the microscope to:

 Reduce the intensity of light

 Increase contrast and resolution

 Adjust the color balance of the light to give the best

visual effect
 Provide monochromic light

 Absorb light

 Transmit light of selected wavelength

 Protect the eye from injury caused by ultra-violet light.47

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2.2.2.3 Working principle of the microscope
 The magnified image of the object (specimen) is first
produced by a lens close to the object called the objective.
 This collects light from the specimen and forms the primary
image.
 A second lens near the eye called the eyepiece (ocular)
enlarges the primary image converting it into one that can
enter the pupil of the eye.
 The magnification of the objective multiplied by that of the
eyepiece gives the total magnification of the image seen in
the microscope.

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See the following example:
 Objective Eyepiece Total
Magnification Magnification Magnification
 10X 10X 100X
 40X 10X 400X
 100X 10X 1000X

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 Objectives
 Low power (10X) Objective
 Used for the initial scanning and observation in most
microscopic work.
 When using 10 X

 Close iris diaphragm

 Lower the condenser
 High -dry power (40X) Objective
 Is used to study un stained specimens such as stool
and urine sediments for more detailed examination.
 When using 40 X

 open the iris diaphragm half way

 raise the condenser half way
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 Oil immersion (100X) Objective
 Routinely used for morphologic examination of blood
films and microbes
 An oil immersion lens requires that special grade
of oil (immersion oil) be placed b/n the objective
and the slide
 The oil is used to increase the intensity of light.
 When using 100 X

 open the iris diaphragm completely

 raise the condenser completely

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2.2.2.4 Resolving power of the microscope
 Itmay be defined as the ability to level closely adjacent
structural details as being actually separate and distinct.
 The increase in magnifying power is always linked to an
increase in resolving power.
 The higher the resolving power of an objective, the closer
can be the fine lines or small dots in the specimen which the
objective can separate in the image.
 The resolving power of an objective is dependent on what is
known as the numerical aperture (NA) of the objective.

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Resolving power….
 The numerical aperture is a designation of the amount of
light entering the objective from the microscope field, i.e. the
cone of light collected by the front lens of the objective (an
index or measurement of the resolving power).
 It is dependent on the diameter of the lens and the focal

length of the lens.

E.g. Res. power of:

 Human eye- 0.25 mm

 Light microscope- 0.25µm

 Electron microscope- 0.5 nm

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 Numerical Aperture
 Defined as the product of the refractive index of the medium
outside the lens (n) and the sine of half the angle of the cone
of light absorbed by the front lens of the objective (u) or

 Is a number that expresses the ability of a lens to resolve

fine detail in an object being observed
E.g. 0.25 on X10 objective
0.65 on X40 objective
1.25 on X100 objective
56
 The
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greater the N.A the greater the resolving power.
 The following are the usual numerical apertures of
commonly used objectives.
 10 X objective ----------- NA 0.25

 40 X objective ----------- NA 0.65

 100 X (immersion oil) objective ------- NA 1.25

 Total magnification
 is the product of the objective and the eye piece magnification
 Useful magnification range
 is
calculated as:
(500-1000)x NA of that objective
E.g. The useful magnification range when an Eyepiece with
magnification of 10x & an objective with magnification 40x & NA
of 0.65 is: 325-650
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Focal length……
 Small Diameter
 Long focal length
 Very low NA
 Very low r.p
 Very low useful magnification
 Small diameter
 Short focal length
 Low NA
 Low resolution
 Low useful magnification
 Therefore the wider the angles of the cone of light the higher
the NA of the objective and greater is the objectives
resolving power and useful magnification.

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2.2.2.5 Working principle of an oil immersion objective
 When a beam of light passes from air into glass it is bent
and when it passes back from glass to air it is bent back
again to its original direction.
 This has effect on oil immersion objective and affects the NA
of the objective and consequently it’s resolving power.
 The bending effect on the objective can be avoided by
replacing the air between the specimen and the lens with oil,
which has the same optical properties as glass, i.e.
immersion oil.
 The oil provides better resolution and a brighter image by
collecting extra oblique light.

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2.2.2.5 Routine use of the microscope
 A microscope must always be used with gentleness, care and
the following should be noted.
1. Place the microscope on a firm bench so that it does not
vibrate.
a. Make sure that it is not be exposed to direct sun light.
b. The user must be seated at the correct height for the
convenient use of the microscope.
2. Select the appropriate source of light.
3. Place the specimen on the stage, making sure that the
underside of the slide is completely dry.

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 Routine…..
4. Select the objective to be used.
 It is better to begin examination with 10x objective.
5. Bring the objective as close as possible to the slide preparation
6. Adjust the light source until the illumination of image is at its
brightest.
7. Focus the condenser.
8. Adjust the aperture (opening) of the condenser iris according to
the specimen being examined.

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Routine…..
 The wider the condenser aperture, the brighter will be the
specimen and the smaller be the details, which can be
resolved.
 The smaller the aperture, the greater will be the contrast.
 Certain specimens, example stained and mounted
specimens give little glare illuminated image with fine detail.
 Other specimens, like urine, unstained cerebrospinal fluid
and saline mounted fecal specimens give much glare and
require a reduced source of light to increase contrast.

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Routine…..
9.Examine the specimen by systematically moving the slide with
the mechanical stage.
N.B: The image of the specimen will be up side down and will
move in the opposite direction to the side.
10. For a higher magnification, swing the 40x objective into place.
 Focus the 40x objective, using the fine adjustment.
 If for any reason the image is not visible, lower the objective
until it is nearly but not quite touching the specimen.
 Then looking through the eyepiece, focus up wards with the fine
adjustment until the image comes into view.
11.For the highest magnification, add a drop of immersion oil to
the specimen and swing the 100x oil immersion objective into
place, then open the iris fully to fill the objective with light.
Example.
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2.2.2. 6 Care, Cleaning and Repair of microscope
 The microscope is one of the most expensive and delicate
instruments.
 Good microscopy practice:
I. Daily cleaning and quality control(QC) check
a. Using a clean cloth, wipe any dust from stage and other surfaces of
microscope
b. Using lens tissue clean dry objective.
 Clean 100X objective with tissue dampened with xylene.
 Never use alcohol to clean the oil because it will dissolve the
cement holding the lens.
c. Carry out a QC check to ensure the lenses are completely clean.

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II- Care when using the microscope
1. Do not force any mechanism.
2. Check stage and under side of the specimen re DRY and
CLEAN.
3. Cover wet preparation with cover slip.
4. Use non-drying oil immersion.
5. Put eyepieces that are mot in use in closed container.
6. Always lift and carry the microscope well supported with
hands.
7. Protect the microscope from dust, moisture and direct
sunlight.

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III- At the end of the Day’s
 Turn the switch off.
 Clean using a soft tissue.
 Do not leave the objective of eyepiece open.
 Decontaminate the stage with 70% alcohol dampened cloth.
 Cover with its dust cover.

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2.2.2: Equipment for purifying water
2.2.1: DISTILLER
 A process by which impure water is boiled and the steam
condensed on cold surface (condenser) to give pure distilled
water is called distillation.
 The apparatus is called distiller.
 Distilled water is free from dissolved salts and clear
colorless, odorless and tasteless. It is sterile too.
 A considerable volume of cool running water is required to
operate or to condense the steam.

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2.2. 2: DEIONIZER
 Deionizer is an apparatus used to produce ion free water.
 A deionizer is an apparatus for demineralizing water by
means of cartridges filled with ion-exchange resin
 Deionizartion is a process in which chemically impure water
is passed through anion and cation exchange resins to
produce ion free water.
 Deionized water has low electrical conductivity, near neutral
pH and is free from water-soluble salts but is not sterile.

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2.3: Equipment for weighing
Balances
 Balances are essential laboratory instruments that are widely
used for weighing of various substances (powders, crystals
and others) in the laboratory.
 For instance, to prepare reagents, stains and culture media,
balances are required to weigh accurately and precisely
within the needed range.
 They should be kept carefully clean and located in an area
away from heavy traffic, large pieces of electrical equipment,
and open windows.
 To minimize any vibration, as interference that may happen,
a slab of marble is placed under the balance.
05/15/24 71
Balances in medical laboratory may be:
A. Rough balances (mechanical balances)
B. Analytical balances/electrical/
2.3.1 Rough balances
 Rough balances are several types. Some of them use sliding
scale, some have a single or double pan (s) and others
utilize dial - operated fractions.
 They are used for weighing substances, which do not call for
extreme accuracy.
 While operating, they do not require mains electricity or
battery power and are currently less expensive than
analytical balances of the similar sensitivity.
 Some rough balances weigh accurately to 0.1 gm of a 72
05/15/24
 Two - pan balance is a rough balance, which has two copper
pans supported by shafts.
 It is used:

 To weigh large amounts (up to several kilo grams)

 When a high degree of accuracy is not required.
 The sensitivity of a two pan balance is 0.5 gm.

 The sensitivity of a balance is the smallest weigh that moves

the pointer over one division of the scale.
 For routine laboratory purposes the sensitivity of a balance
can be considered to be the smallest weigh that it will
measure accurately.
 Usually the larger the amount of substance to go into a
reagent, the least accuracy is required.
05/15/24 73
 2.3. 2 Analytical balances
 Nowadays analytical and electronic balances (single pan
balances that use an electron magnetic force instead of
weights) are the most popularly used balances in medical
laboratories to provide a precision and accuracy for reagent
and standard preparation.
 Analytical balance is a highly sensitive instrument.
 It may have two pans suspended from a cross beam, inside
a glass case.
 It requires mains electricity or battery (D.C) supplied power.

05/15/24 74
 These balances are used:
1. To weigh small quantities usually in mili gram
(mg) range
2. When great accuracy is required. E.g., 2.750mg,
0.330 mg, 5.860mg, etc
 Its sensitivity is 0.5 mg to 1 mg depending on the model.
 N.B: The accuracy of a balance should be checked regularly
as recommended by the manufacturer.

05/15/24 75
05/15/24 76
 Use and care……..
 When adding or removing a chemical, remove the container to avoid
spilling any chemical on the balance.
 When using an analytical double pan balance, bring the pans to rest
before adding or removing a chemical.
 Always use forceps to add or remove weighs. Protect the weights
from dust, moisture and fungal growth.
 Use small brush to remove any chemical, which may have been spilt
on the balance.
 A container of self - indicating silica gel should be kept inside the
analytical balance case to remove any moisture present in the
atmosphere.
 Keeps the balance clean, being particularly careful not to let dirt
accumulate near the pivots and bearings.

05/15/24 77
05/15/24 78
2.2.4: Equipment for pipetting and dispensing
 There are different types of devices used for pipetting and dispensing
specimens. Some of them are:
 Simple bulb aspirator- this is simple inexpensive device suitable
for use with graduated capillaries.
 Thumb wheel aspirator – it can be used with capillaries, shell-
back pipettes, example, Sahli or WBC pipettes ad most small bore
graduated pipettes, example measuring up to 0.5ml.
 Automatic pipetter – it use plastic or glass tips and models are
available for measuring single volumes or several different
volumes. Automatic pipetters have a greater precision and
accuracy.
 PVC bulb pipette filler – its tapered and flexible end enables all
types of pipettes up to 10 ml volume to be inserted easily and
safely in to the end and to be held securely.

05/15/24 79
Equipment for pipetting and ……

 Pi- pump2500, pipette filler – it is highly recommended for the controlled

filling and dispensing of fluid from pipettes.
 Bottle top dispenser - it is used to measure a fixed volume of fluid or
several different volumes of fluid.
 Bottle top hand operated dilutor – this is the most expensive of the
devices described above. It is used for measuring accurately and precisely,
specimen and reagent.
 Plastic bulb pipettes – Plastic bulb pipettes have many uses in a medical
laboratory. They can be decontaminated in disinfectant, wash, and reused
many times.

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2.2.5: Laboratory centrifuges
 Centrifuge: is equipment that is used to separate solid
matter from a liquid suspension by means of centrifugal
force.
 They sediment particles (cells, bacteria, casts, parasites,
etc.) suspended in fluid by exerting a force greater than that
of gravity.
 The suspended materials are deposited in the order of their
weight.
 There are many types of centrifuges, but the basic principle
is the same, that is, the all use centrifugal force.
 When a body is rotated in circular movement at speed,
centrifugal force is created that drives the body away from
the center of the circular movement.
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Centrifuge….
 The greater the outward pull due to rotation, that is
centrifugal force, the more rapid and effective is the
sedimentation.
 As a result, heavier elements are thrown to the bottom of the
tube followed by lighter particles.

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Centrifuge….
 Centrifugal force increases with the speed of rotation that is
the revolution of the rotor per minute and the radius of
rotation.
 The actual sedimentation achieved at a given speed
depends therefore, on the radius of the centrifuge.
 Most techniques requiring centrifugation will usually specify
the required relative centrifugal force (RCF) expressed in
gravity.

05/15/24 83
2.2.5.1 Basic components of centrifuges
 Central Shaft: - It is a part that rotates when spinning is effected
manually.
 Head: - It is a part that holds the bucket and connected directly to the
central shaft or spindle.
 Bucket or tube: - Are portions that hold test tubes containing a given
sample to be spined.

Specimen
Tube

Cup

Shaft
05/15/24 84
2.2.5.2 Classifications of centrifuges
A. Hand centrifuges
 These centrifuges are:

 Operated by hand or water pressure and they are

most commonly used in small laboratory for routine
purposes,
 Used for preparation of urinary sediments and to concentrate
parasites from the given specimen and it is not advisable to use
them to separate serum from whole blood.
B.Electrical Centrifuges
 Electrical centrifuges are those centrifuges that are operated
by electrical power and produce high centrifugal force.
 They are used in most medical laboratories.

05/15/24 85
 Based on their tube angle rotation, there are two
types.
A. Swing out head:- This is the most frequently used type and
the head is designed to swing the tubes to the horizontal
position during centrifugation process.
B. Fixed head: - They have different angles. They are useful for
certain laboratory techniques. Example, for agglutination tests
in blood grouping using test tube method.
2. 2 .5. 3 Kinds of centrifuges
1. Micro-centrifuges
 They are used for spinning small tubes as in blood
bank laboratories.

05/15/24 86
2. Medium size centrifuges.
 Are used for centrifuging of urine specimens for microscopic
analysis of urinary sediments.
3. Large centrifuges
 They are widely applied in bacteriology and medical
chemistry laboratories.
 A centrifuge may have a preset speed or more often there is
a knob by which the laboratory personnel control the speed.
 The speed is given in revolution per minuets (rpm).
 Small models are designed to centrifuge volumes up to 200
ml at maximum speeds of 3,000 - 4,000 rpm.
 Large models will centrifuge volumes up to 2,200 ml with
maximum speeds of 5,000 rpm.
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  A centrifuge may have built in timer or may have to be timed
with a watch. Some centrifuges may have a temperature
gauge in order to keep the temperature constant as it
spines.
4. Cyto-centrifuge
 Specific use
 Spreading of cells across slide
 Body fluids
 Microscopic – morphologic slides

5. Ultracentrifuges
 High-speed
 Up to 90,000 – 100,000 rpm; 178,000 g
 More common in research

05/15/24 88
2.2.5.3 Use and care of centrifuges
 Although most centrifuges are fitted with an imbalance
detector, lid interlock, and an automatic braking system, it is
important for laboratory workers to know how to use a
centrifuge correctly to prevent it from damage and
breakages.
 These include:

 Reading the manufacturer’s instructions.

 Placing a centrifuge on a firm level bench out of direct

sunlight, towards the back of the bench.
 Whenever possible using plastic tubes made from
polystyrene or autoclavable.

05/15/24 89
 Always closing the centrifuge top before turning it on.
 Always balancing the tubes that are being centrifuged.
 Tubes of the same weight should be placed directly opposite to
each other.
 Tubes should also be of the same size and should also
contain the same amount of liquid.
 Increasing spinning speed gradually is important.
 Give the centrifuge a chance to come up to that speed and
then turn up the dial a little further until it reaches the desired
3,000 rpm.
 Five minutes are the usual time required to centrifuge most
substances.

05/15/24 90
 Never open the centrifuge while it is still spinning. Never try
to slow it down with your hand. Most centrifuges have a
brake, which will cause the centrifuge to stop faster.

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2.2.6: laboratory autoclaves, ovens
2.6.1 AUTOCLAVE
 Autoclave is an instrument that operates by creating high
temperature under steam pressure.
 Autoclaving is the most common, effective, reliable and
practical method of sterilizing laboratory materials.
 Temperature of 1210c, which will kill spores with in 15
minutes and at 15 psi /pound/.
 At this particular temperature, pressure and time, all forms of
lives are destroyed.

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05/15/24 93
 Precautions in the use of autoclaves
 The following guidelines can help to minimize risks while
working with autoclaves.
1. Proper use and care of autoclaves.

2. Regular inspection of the chamber, door seals and

gauges.
3. The steam should be saturated and free from chemicals
that could contaminate the items being sterilized.
4. Materials to be autoclaved should be in containers that
allow ready removal of air and permit good heat
penetration.
5. The chamber of the autoclave should be loosely packed
so that steam will reach the load evenly.
05/15/24 94
 6. Operator should wear protective gloves for protection when opening
the autoclave.
7. Thermocouples should be placed at the center of each load in order to
determine proper operating cycles.
8. Ensure that the relief valves of pressure cooker autoclaves do not
blocked.

05/15/24 95
2.6.2 OVENS
 Hot - air ovens are instruments that are used for drying of
chemicals and glass wares.
 They are also used for the sterilization of various glass
wares and metal instruments.
 They consist of double walls that are made of copper or
steel.
 They are heated by circulation of hot air from gas burners
between the metal walls or by electrical mains.
 There is a thermometer on the top of the ovens and
generally an automatic device (thermostat) is fitted to
regulate the temperature.

05/15/24 96
05/15/24 97
2.2.7: Incubator and water bath.
2.2.7.1 INCUBATOR
 Incubation at controlled temperature is required for bacteriological
cultures, blood transfusion, Serology, Hematology and clinical
Chemistry tests.
 For bacteriological cultures, an incubator is required whereas for
other tests a dry heat block or a water bath may be used.
 For the incubator, the air inside is kept at a specific temperature
(usually at 370c). When tubes are kept inside the incubator, they take
the temperature of the incubator.
 The appropriate temperature is obtained by means of temperature
regulator and is maintained by a thermostat. This permits a more
accurate temperature control.

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 Use and Care of Incubator
 Read carefully the manufacturer’s instruction.
 Make sure the incubator is positioned on a level surface and
that none of the ventilation openings are blocked.
 If the incubator does not have a temperature display, insert a
thermometer in the vent hole through the roof of the
incubator. Adjust the thermostat dial until the thermometer
shows the correct reading, i.e., 35 - 37Oc for the routine
incubation of bacteriological cultures.
 Before incubating cultures and tests, check the temperature
of the incubator.
 Clean the incubator regularly; making sure it is disconnected
from its power supply.
 Every three to six months check the condition of the
incubator.
05/15/24 99
 At the time of purchase, it is advisable to buy a spare
thermostat and thermometer if these are of special type and
are not available locally.

05/15/24 100
2.2.7.2 WATER BATH
 The water bath, like the incubator, is required for controlled
temperature incubation of culture and liquids, and many
other laboratory tests.
 The temperature of the water bath is thermostatically
controlled and can be set at any desired level ranging
usually from 20oC to 100oC.
 The heating coil may be of immersion type or enclosed in a
case, some models have propellers to help to circulated
water so that identical temperature is maintained throughout
the water bath

05/15/24 101
 Use and care of water bath
 Maintain the minimum level in the water bath with chemically
pure water. Avoid use tap water. Avoid use of water as salts
from tap water may get deposited on coil and so affect its
function
 Always use a thermometer to check that the temperature is
stable at the desired level.
 Make sure that the substance being incubated is below the
surface of water in the bath.
 It is advisable to cover the tubes, flasks or plates during
incubation to avoid contamination and dilution as a result of
condensation of water from the lid of the water bath.
 Clean the water bath regularly

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2.2.8: Colorimeter
 Colorimeter (Photometer)
 Colorimeter is an instrument used to measure the

concentration of a substance in a sample by comparing

the amount of light it absorbs with that absorbed by a
standard preparation containing a known amount of
the substance being tested.
 In a test a colored solution of the substance being

measured or a colored derivative of it is produced this is

measured in a color meter colored solutions absorb light
at a given wavelength in the visible spectrum.
 Biological samples contain many substances that can be

determined quantitatively or qualitatively.

05/15/24 103
Basic components of a colorimeter
 A constant source of radiant energy
 Some optics for focusing the light
 a colored filter
 a cuvette holder
 Light- sensitive detector( Converts light energy to electrical
energy)
 Read out device

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Spectrophotometer
 Is similar to a colorimeter except that instead of using a
filter to select the colour of the light to pass through the
sample, the white light is separated into a
rainbow( spectrum of colours) using a prism or diffraction
grating
 Continuous adjustment of λ with the help of prisms or

diffraction gratings.

05/15/24 105
2.2.9: Mixers: -
 are instruments used for preparation of reagents for mixing or
dissolving purpose.
 Also used for homogenization.

05/15/24 106
2..2.10: Refrigerators
 Refrigerators are physical means of preserving various
laboratory specimens.
 They suppress the growth of bacteria and maintain the
specimens with little alteration.
 In addition to this, they are also used in the medical
laboratory to preserve some reagents such as:
 Pregnancy tests kits,
 Rapid plasma reagin (RPR) test kits,

 Culture media are also preserved

 Blood grouping anti sera and others which

are kept in the refrigerators to prevent their

deterioration.
 Figure

05/15/24 107
2.2.11: Desiccators
 Desiccators are instruments, which are used for drying of
chemicals or to keep other chemicals from being hydrated.
 As chemicals stay for long period of time out of dessicators,
they sometimes absorb water
 The chemical is dried in an oven at 110oc for 1 hour, and then
it is placed in a desecrator over night before weighing on the
analytical balance.
 The purpose of the oven is to remove the water and that of
the desicator is to store the chemical at an ambient
temperature where it cannot reabsorb water.

05/15/24 108
Desicator…….
 A desicator contains substances called drying agents.
 These absorb the water in the air of the desicator.
 The most commonly used drying agents (desiccants) are
calcium chloride and concentrated sulfuric acid.
 The chemical that is to be dried is placed in another bottle or
test tube and is put on top of the desiccants present in a
securely closed desicator.

05/15/24 109
2.2.12 PH meter
Definition: is an instrument which is used to measure
Potential of ion hydrogen (i.e. acidity or alkalinity of a
substance) or Is an instrument used to measure the PH
or H+ ion concentration.
 Potential of hydrogen pH scale is 0 – 14
 Acid pH: 0-6.9
 Neutral pH: 7.0
 Alkaline pH: 7.1-14.0

05/15/24 110
Major component
1. Glass bulb electrode( PH- electrode)
2. Reference( Calomel) electrode
3. Potentiometer (Sensitive meter) which measures the
electric volt.
 The glass bulb electrode contains a solution of a certain

fixed PH or H+ conc.
 When the electrodes are placed in a solution of unknown

PH, an electrical potential is produced between them( i.e

the solution and the H+ ions in the PH-electrode)

05/15/24 111
Major parts…….
 This potential which is proportional to the H+ ion
concentration of the test solution, is measured with the
aid of reference electrode which is compared to the
potential of the PH-electrode.
 The mili volt(MV) potential difference is displayed as

digital or galvanometric readings(PH0-14) OMV=7.0

05/15/24 112
2.2.13: Safety cabinets
 Safety Cabinets are designed to protect the laboratory
personnel, the laboratory environment and work materials
from exposure to infectious aerosols and splashes that may
be generated when manipulating materials containing
infectious agents, such as primary cultures, stocks and
diagnostic specimens.
 These cabinets could be chemical or biological
 N.B: It is extremely important to use gloves as a personal means of
protection from various infectious agents while working in medical
laboratories.

05/15/24 113
Biological safety cabinets/BSC/
 Are the principal equipment used to provide physical
containment
 Are used as primary barriers to prevent the escape of
aerosols into the laboratory environment.
 Certain BSC can also protect the test/specimen from a ir
born contamination
 The selection of BSC is based on:
 The hazard of the agent in the test
 The potential of the the laboratory technique to produce aerosols and
 The need to protect the test from airborne contamination
 Three types of BSC are available- Class I ,Class II, Class III

05/15/24 114
Class I –BSC
 This is an open fronted work chamber which is exhaust
ventilated to provide personnel and environmental protection
only by means of an inward air flow away from the operator,
the exhaust air being filtered through a HEPA filter before
being discharged from the cabinet
 Are used with agents with low to moderate risk

 Class II- BSC

 Is a partially open fronted work chamber which provides
protection for personnel and the surrounding environment
against biological hazards by means of barrier airflow at the
work opening.
 A quantity of air equal to the barrier air is exhausted from the
cabinet through a HEPA filter.
05/15/24 115
Class II…
 Also provides product(test) protection against contamination
by means of HEPA filtered air flowing into the cabinet
 Class III-BSC
 Is a totally enclosed and gas-tight structure
 Work procedures in the cabinet are carried out through
replaceable arm length gloved sleeves.
 Is supplied with air through a HEPA filter and exhausted
through two HEPA filters mounted in series
 Suitable for use with all categories of biological agents

 HEPA- high efficiency particulate air

05/15/24 116
Safety cabinets

05/15/24 117
2.3: Care and cleaning of laboratory equipments and wares
 Cleaning of glasswares
 It is clear that volumetric glasswares and glass apparatus
must be absolutely clean, otherwise volumes measured will
be inaccurate and chemical reactions are affected adversely.
 One gross method generally used to test for cleanness is to
fill the vessel with distilled water and then empty it and
examine the walls to see whether they are covered by a
continuous thin film of water.
 Imperfect wetting or the presence of discrete of droplets
water indicates that vessel is not sufficiently clean.

05/15/24 118
Cleaning of ….
 A wide variety of methods have been suggested for the
cleaning of most glassware.
 Wide varieties of methods have been suggested for the
cleaning of most glassware.
 In all cases, glassware for the clinical laboratory must be
 Physically clean

 chemically clean

 bacteriologically clean or sterile

05/15/24 119
Cleaning of new glassware
 New glass ware should be appropriately treated and
cleaned before use.
 Newly purchased soft glass ware (soda lime) should be
treated overnight with 5% HCl.
 This will neutralize free alkali found on the surface.
 This treatment is not necessary for borosilicate glass
(hard glass).
 Acid treated glassware must be first rinsed with tap water
followed by through rinsing with distilled deionnized
water.
 Newly purchased borosilicate glass ware is cleaned with
detergent followed by washing with tap water and then
rinsing with distilled water.
05/15/24 120
General cleaning procedure
1). Preliminary rinsing
 Rinse all glassware immediately after use. Remember, dry
glassware, like the dry dishes after a meal, is difficult to
clean, stains, markings, proteins and other materials may
get stubborn due to drying.
 Rinse twice in cold or warm water.
2) Soaking in deteregent solution
 Place in detergent solution (2%).Detergents can be in either
solution or powder form. Do not use too concentrate
detergent solution which may not be completely removed
and this will affect the test result.
 Dissolve the detergent completely before putting in the
glassware.
05/15/24 121
General. .….
 Preliminary soaking in the detergent can save time, reduce
contamination problems and make the final washing greatly
simplified. Soak for one hour.
3).Scrubbing
 Scrub thoroughly with good quality brush (choose
appropriate brush for the type of glassware being cleaned). A
milled abrasive may help cleaning but the abrasive should
not scratch the glass.
 Make sure that the brush reaches all parts of the glassware, inside and
the out side.
4). Washing
 Wash each glassware one by one under running water.
Wash each item 5 times or more.
05/15/24 122
General ……
5). Rinsing
 Rinse each glassware with distilled water or deionnized
water at least three times.
6). Drying
 Place in a wire baskets and dry glassware completely by
keeping it in an oven (1400c)If an oven is not available, dry
the glassware on the drying rack at room temperature over
night.
 Dry the burette in the inverted position on the burette stand.
Glassware dried in the hot air oven should be in an inverted
position to ensure complete drainage of water as it dries.

05/15/24 123
General……
7). Plugging
 The clean dry glassware should be put away in a cup board
to protect it from dust.
 It is recommended that containers should be plugged with
non – absorbent cotton wool or the mouth covered with little
cups made from wrapping paper or preferably thin sheeting
of paraffin wax.

05/15/24 124
1. Cleaning of pipettes
 Pipettes should be placed in a vertical position with the tips
up in a jar of cleaning solution in order to avoid the breakage
of their tips. A pad of glass wool is placed at the bottom of
the jar to prevent breakage.
 After soaking for several hours, the tips are drained and
rinsed with tap water until all traces of cleaning solution are
removed.

05/15/24 125
 The pipettes are then soaked in distilled water for at least an
hour.
Filing with water, allowing the pipette to empty, and observing
whether drops formed on the side within the graduated portion make
a gross test for cleanness.
 Formation of drops indicates greasy surfaces after the final distilled

water rinse the pipettes are dried in an oven at not more than 110 oc.
 Most laboratories that use large numbers of pipettes daily use a
convenient automatic pipette washer.
 These devices are made of metal or polyethylene and can
be connected directly to hot and cold water supplies.
Polyethylene baskets and jars may be used for soaking and
rinsing pipettes in chromic acid cleaning solution.

05/15/24 126
2. Cleaning of flasks, beakers, cylinders and other glass
wares
 Pour warm cleaning solution into each vessel and stopper or
cover carefully.
 Each vessel should be manipulated so that all portions of the
wall are repeatedly brought into contact with the solution.
 This procedure should be followed for at least five minutes.

05/15/24 127
 The cleaning solution can be poured from one vessel to
another and then returned to its original container.
 The vessels should then be rinsed repeatedly with tap water
four times and finally rinsed three times with distilled water.
 It is important that the necks of volumetric flasks above the
graduation mark be clean because when solutions are
diluted in the flask drops of water may adhere to an unclean
wall and may invalidate the measurement of volume.

05/15/24 128
3. Plastic wares:
 Plastic wares are usually manufactured from polymers of
polyethylene, polypropylene and TEFLON.
 These plastics are chemically inert and unaffected by
acid /alkali.
4. Cleaning of plastic wares
 After each use Laboratory plastic wares should be
immediately soaked in water or if contaminated soaked
overnight in a suitable disinfectant such as 0.5% w/v sodium
hypochlorite or bleach.
 Most plastic ware is best clean in a warm detergent solution
followed by at least two rinses in clean water and ideally a
final rinse in distilled water.

05/15/24 129
 The articles should then be left to drain and dry naturally or
dried in a hot air oven, set at a temperature the plastic can
withstand.
 A brush or harsh abrasive cleaner should not be used on
plastic ware.
 Stains or precipitates best removed using dilute nitric acid or
3% v/v acid alcohol.

05/15/24 130
Next: chapter 3: Collection, handling and shipment
of laboratory specimens
ALL!!!!!!!!!!

05/15/24 131