SAFETY IN THE

HISTOPATHOLOGY
LABORATORY
CECILIA LEKPOR

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 INTRODUCTION
Laboratory safety involves measures to protect against laboratory accidents
 Includes safety training and enforcement of laboratory safety policies
safety review of experimental designs and the use of personal protective
equipment
 It is the responsibility of every person working in the medical laboratory to
ensure a safe working environment and not threatening the safety of co-workers
and anyone who enters the laboratory

Development of skills and responsibility must be an integral part of every

Laboratory set up
You need to read the safety manual, which sets out the hazards and safe working
procedures
1/21/2022 for the laboratory
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INTRODUCTION
 workers in pathology laboratories face hazards such as:
 Specific risks from toxic chemicals which are used in investigative procedures e.g
xylene, formaldehyde, alcohols etc
 Pathogenic microorganisms in patient's samples e.g body fluids
 General risks from mechanical, electrical and fire hazards which is worsen by:
 Ignorance of the hazards,
 Lack of knowledge on safety measures and inadequate safety measures adopted
by laboratories in order to identify and evaluate the hazards

Most hazards encountered fall into three main categories: chemical, biological,
or physical.

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 PHYSICAL ENVIRONMENT
 Organisation of the physical environment of the Laboratory promote safety.
 A clean, tidy laboratory offers less chance of an accident occurring
CONTAINMENT SYSTEMS
Containment systems are designed to contain harmful substances at source,
thereby preventing their release into the laboratory

This area separates the hazard from contact with laboratory workers and with
the general environment within the laboratory
You need to know the various containment systems, e.g primary, secondary and
tertiary
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PERSONAL PROTECTION
Protective clothing of an approved design must be worn by all students within the
laboratory work area and removed before leaving the laboratory work area, e.g. gowns,
coveralls, aprons, gloves etc.

Personal hygiene is very important and therefore must be strictly adhered to:

 NO eating, drinking, smoking or the application of cosmetics in laboratory work

areas nor in any area where workplace materials are handled

 NO food and drink to be stored in the laboratory (may be stored in the rest area)

 Regular hand washing at the end of each job or activity session immediately after
handling chemicals and specimens and before leaving the laboratory
 Wearing of pendant jewellery not permitted (advise no wearing of any jewellery
other than wedding rings)

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PERSONAL PROTECTION CONT’
Safety spectacles, goggles and visors must be use
Goggles give better protection than safety spectacles for work with
hazardous chemicals
Mouth pipetting must be forbidden
 Hypodermic needles must be used instead of pipettes.
 Cracked and chipped glassware must always be discarded and not
reused.
Specimens must be received in a safe condition. Specimens should be
unpacked with care and attention to possible breakage and leakage.
 Gloves must be worn for unpacking specimens.
 Work benches must be kept clean and tidy.
 Appropriate disinfectants must be used correctly
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SAFETY IN THE HISTOPATHOLOGY LAB
EQUIPMENT:
 All equipment is potentially dangerous, do not use it until you have been
properly be shown how to use them. Eg Microtome, Embedding machine,
Tissue processor etc.

Report to a senior personnel if an equipment is not working properly.

Do not open the centrifuge until the rotor has stopped.

Unplug any electrical equipment before you try to open it

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SAFETY IN THE HISTOPATHOLOGY
LAB-CONT’
Unplug any electrical equipment before you try to open it

Faulty design or construction may lead to electrical fires or

shock as a result of:
• Improper use of equipment
• Improper adaptation
• Lack of proper maintenance
Always consult a senior personnel before using any equipment
in the Histology Lab
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 REAGENTS/CHEMICALS
All chemicals present some degree of danger because of toxicity,
corrosiveness, irritation and/or flammability
Toxicity and irritation can be caused by skin contact, ingestion or
inhalation of a solid, liquid or vapour
The toxic effects of chemicals can be classified on the basis of their ability
to cause specific types of disease processes:
Allergens: chemicals recognised by the immune system as antigenic and
cause hypersensitivity reactions – contact dermatitis, asthma
Carcinogens: chemicals that cause or increase the risk of cancer

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REAGENTS/CHEMICALS CONT’

Reproductive toxins: include mutagens, chemicals that can

induce mutations of the germ cells resulting in genetically-
induced malformations, spontaneous abortions or stillbirth, or
death of offspring of the exposed person

Teratogens: chemicals that can damage the foetus, causing

congenital malformations or death

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REAGENTS/CHEMICALS CONT’
Do not carry reagents and chemical bottle by the neck. Containers
should be transported on carriers and never by the neck

Plastic aprons should be worn on to protect from splash when working

with or around formaldehyde, preparing solutions and working around
fresh unfixed tissue or body fluids.
Wear gloves when the potential of contact with toxic material exist

Wear disposable sleeve garments to protect arms of contact with

biohazards and chemicals.
Wear goggles for eye protection when preparing solutions, grossing
specimen, changing solutions on equipments.
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REAGENTS/CHEMICALS CONT’
Work in Laboratory hood to prevent hazardous vapour from entering
the general Laboratory area.

All chemicals should be labelled with approved warning labelling

scheme to help in identifying hazards

The label should include the chemical and common names,

warnings about physical and health hazards, advice on safety
precautions, and name and address of manufacturer

The label may also contain signs, symbols and pictures depicting
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 HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS

 FORMALDEHYDE: Formalin is typically 37% formaldehyde by weight

(40% by volume) and 6-13% methanol by volume in water
 The permissible exposure limit (PEL) for formaldehyde in the workplace
is 0.75 parts formaldehyde per million parts of air (0.75 ppm) measured as
an 8-hour time-weighted average (TWA)
 All workers exposed to formaldehyde should be monitored for exposure
levels on a periodic basis Increased risk of leukemia particularly myeloid
leukemia and brain cancer compared with the general population
 Some studies showed association with nasopharyngeal cancer , others
did not show any association with lung cancer
You1/21/2022
need to be mindful of this
SAFETY when using
IN THE HISTOPATHOLOGY formalin
LABORATORY-Cecilia Lekpor solutions 13
 HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT’
Drain disposal of limited quantities of formaldehyde may be permitted in
some communities
 Recycle as much waste as possible by distillation and have the
remainder taken away by a licensed waste hauler or detoxified by a
commercial product.
 Formaldehyde is readily biodegradable. Nearly all organisms have an
enzyme, formaldehyde dehydrogenase, which decomposes this chemical

 The trick is to feed it into the system slowly enough so that it is diluted
below toxic concentrations by the normal flow of water.
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HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT’
For small/minor formaldehyde spills, clean surfaces with cold water at
least two times.
 If absorbents are used, place them into a plastic bag and seal the bag.

Preparation of formaldehyde containing solutions:

 Potential over-exposures can occur when preparing paraformaldehyde
and 35% formaldehyde solutions

 Prepared in chemical fume hood to prevent exposures and to contain

spills If a spill occurs within the fume hood, the spill can be considered
minor and can be cleaned up using paper towels.
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HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT’

 Place the aldehyde contaminated paper towels into a sealable

bag for disposal as hazardous waste

Clean all equipment after use and the working area

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 HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT’
ALCOHOLS: ETHANOL
Not a serious health hazard under most conditions of use
 Inhalation may cause dizziness, headache and irritation to respiratory
passages
Skin contact can cause excessive drying and dermatitis
 ACETONE Highly flammable (flash point = −16°C) and very volatile
Great risk of fire from heavy vapors traveling along counters or floors to a
distant ignition source
 STORE using tight-fitting lid and store in a space where there are no
electrical outlets, stoves, or heat-producing sources.
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HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT
XYLENE: Laboratory-grade xylene is composed of m-xylene
(40–65%), p-xylene (20%), o-xylene (20%) and ethyl benzene
(6-20%) and traces of toluene, trimethyl benzene, phenol,
thiophene, pyridine and hydrogen sulfide

 Xylene is a carcinogen, therefore care must taken when

handling

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 HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT
NITRIC ACID: Corrosive to skin, mucous membranes and most metals.
Toxic by inhalation.
Target organ effects on reproductive and fetal systems after ingestion.
 Concentrated acid is very hazardous.
 Use Neoprene gloves for extensive use; nitrile, butyl and latex are not
effective except to protect against minor splashes.
 Wear apron and goggles for handling any quantity.
Always add acid to water, never water to acid, to avoid severe splattering.

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HAZARDS AND HANDLING OF COMMON
HISTOLOGICAL CHEMICALS-CONT
SILVER SALTS AND SOLUTIONS: Skin and eye irritants.
Ingestion will cause violent gastrointestinal discomfort.
 Little risk to workers when fresh, but SOME AGED
SOLUTIONS BECOME EXPLOSIVE.

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BIOLOGICAL (INFECTIOUS) HAZARDS
 Histopathology and cytology laboratory infectious hazard arises from
handling fresh tissues or fluids

All fresh material must be treated as potentially infectious

Disposable aprons and gloves must be used when handling fresh

specimens
unfixed specimens should be handled carefully to avoid aerosol formation
Cysts should be opened gently and cautiously to avoid splashing
Known infected tissues should be handled in a safety cabinet
Instruments and cutting boards must be disinfected after use
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 BIOLOGICAL (INFECTIOUS)
HAZARDS-CONT’
 In the cytology Lab, the risk of infection is created by formation of
aerosols and splashes from fluids

Any manipulation of the specimens from opening of bottles

through filling of tubes for centrifuging to spinning tubes and
sudden braking of centrifuge can create aerosols.

Safety cabinet must be used when handling such specimens

Waste from fresh specimens should be disposed off after

disinfection and in accordance with MoH/GHS guidelines and
Environmental Protection regulations
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DISINFECTION
Disinfectants are used to eliminate virtually all disease causing
microorganisms on objects or surfaces, except bacterial spores, to a level
that is not normally harmful

All disinfectants should be used in the proper dilution as directed by the

manufacturer and should be regularly checked to ensure effectiveness

Centrifuges used in the cytology lab should be cleaned daily with

glutaraldehyde.

If a tube breaks in the centrifuge, time must be given for aerosols to
settle before opening the lid.

 The contents can then be sterilised either by autoclaving or using

activated
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SUMMARY-PRACTICAL SAFETY
PROCEDURES
FIXATION:
Most of the chemicals used in fixation are poisonous
The commonest used fixative is 10% buffered formalin, prepared
from formaldehyde.
 The pungent fumes of formalin affect the eyes, the nose and
throat.
 It can also cause contact dermatitis and is potentially
carcinogenic.
 When using it gloves must be worn and good ventilation is very
essential

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 SUMMARY-PRACTICAL SAFETY
PROCEDURES
DECALCIFICATION
The concentrated mineral acids used for rapid decalcification should
be handled carefully
Tissues fixed in formalin should be well washed before decalcifying
in HCl because of the risk of generating carcinogenic fumes of bis-
cloromethyl ether (when acid mixes with formalin)
FROZEN SECTION TECH:
This involves handling fresh specimens and all precautions on
handling infectious material apply
Cryostat blocks may be frozen using solid CO2 and acetone mixture,
regarded as a fire hazard
Solid CO2 and liquid nitrogen can cause freeze burns. Protective
insulating
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gloves should SAFETY
be INused
THE HISTOPATHOLOGY LABORATORY-Cecilia Lekpor 25
 SUMMARY-PRACTICAL SAFETY
PROCEDURES
TISSUE PROCESSING:
Tissue processing involves the use of flammable chemicals and waxes
on automatic tissue processors.
 High evaporation rates of solvents caused by the method of
transferring tissues from beaker to beaker, and poorly fitting lids in
rotary carries the risk of toxic and potentially explosive build up of
fumes in the environment.
 All electrical plugs and sockets in this room should be made sparkless
to lessen the chances of vapour burning
Wax baths on tissue processors should have safety cut-outs to
prevent over-heating
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SUMMARY-PRACTICAL SAFETY
PROCEDURES
Plastic Embedding
Many of the resins used in plastic embedding for electron
microscopy are toxic and must be handled with care.
 Epoxy resins can cause dermatitis and gloves must be worn during
handling
 Some acrylic resin produce harmful fumes during polymerisation
Staining Procedures
• Many stains are dangerous
• Basic fuchsin (used in Schiff reagent) is regarded as carcinogenic and
should be handled with care to avoid splashing.

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LAB QUESTION

What safety measures will you consider when working in the

in the histopathology Laboratory.

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 THANK YOU

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