Transcript No Slide Title

Essentials of Laboratory Safety
Heather D. Durham, PhD
Chair, Laboratory Safety Committee
Montreal Neurological Institute
Slides adapted from:
Mr. Wayne Wood, Manager ,
Environmental Health and Safety, McGill University
and
Howard Hughes Medical Institute, Online Safety Course
http://www.practicingsafescience.org/
Organization of the Presentation

MODULE 1: General
Laboratory Safety










Getting Ready to Work
Working in the Lab
Emergency Response
Individual Protection
Chemical Hoods
Biosafety Cabinets
Tissue Culture
Using Autoclaves
Chemical Storage
Leaving the Lab

MODULE 2: WHMIS


Classification of Products
Main Elements of WHMIS


Labels
MSDS
You are expected to be familiar with the
information on the website of McGill
Environmental Health and Safety (EHS)
http://www.mcgill.ca/ehs/
Additional resources are listed on the website
for the MNI Laboratory Safety Committee
http://www.mni.mcgill.ca/
MODULE 1: GENERAL LABORATORY SAFETY
All personnel must be aware of general occupational
health and safety issues and procedures, have read the
safety manuals applicable to their work, and completed
the required training.
Laboratory Safety Manual
http://www.mcgill.ca/ehs/laboratory/labsafety/
Biosafety Manual
http://www.mcgill.ca/ehs/laboratory/biosafety/
Radiation Safety Manual
http://www.mcgill.ca/ehs/radiation/manual/
Consult your supervisor to determine the training
required in your laboratory.
Before Lab Work, Get to Know:

Hazards of materials &
agents and their
prescribed safety
procedures

Proper and safe use of
all equipment

First get ready, then get
to work!
Also Get to Know:




Emergency spill procedures,
use of adsorbents and
disinfectants
General Emergency
Procedures for your
Institution
Designated escape route and
alternate
Location of fire
extinguishers, eye wash,
shower, first aid, and spill
kits
Before Lab Work, Get to Know:
Emergency telephone
numbers and reporting
procedures
For all emergencies call:

55-555
While Working in the Lab:

Authorized persons only

Identify EVERYTHING!

No food, beverages, tobacco
products, or application of
cosmetics

Follow Good Laboratory
Practices

Practice Safe Science
Weekly/Monthly Lab Checks
Download file: MNI laboratory safety inspection checklist.xls from MNI website


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Fume Hoods/Biosafety
Cabinets
Tubing, pressurized
connections
Chemical storage
Cleanliness &
Orderliness
Eye wash (purge)
Fire extinguisher
First Aid Kits
Evaluating Lab Hazards




Minor Variation in
Procedure


Regular review of the
types of hazards:
chemical
physical
biological
ergonomic
mechanical
Causes of injury in laboratories
While Working in the Lab:
Report all:
1
Accidents
Injuries
Fires
Spills
Close calls
Disabling injury
10
Minor injuries
30
Property damage incidents
Malfunctioning equipment
Safety hazards
600
Close Calls
Emergency Response
When you take a job in a new laboratory, one of the
first things you should do is ask your supervisor to
review with you the emergency response plans for the
lab. Make particular note of the locations of:

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Emergency telephone numbers.
Eyewash fountains and emergency showers.
Spill kits.
Fire extinguishers.
Emergency exits and evacuation routes.
Fire

Fire is the most potentially devastating
emergency in the modern biology laboratory.
It is imperative that you know how to
prevent fires and be prepared to respond
should a fire occur.

Preventing fires. Use of flammable solvents
is a primary cause of lab fires. Always
follow these prudent practices:



Use the smallest quantities of flammable
solvents as practical.
Store stock quantities in flammables storage
cabinets.
Separate flammable solvents from sources of
ignition.
Never use a Bunsen burner in any area where
flammable solvents are handled.
Emergency Response to Fire




Know the fire emergency response procedures of your
laboratory and institution procedures of your laboratory.
Keep the lab aisles and evacuation routes free of equipment
and other objects that could obstruct safe passage. Remember:
Safe passage is for your benefit and for the benefit of
emergency responders.
Periodically practice emergency response procedures.
Follow these immediate procedures in case of a major lab fire:
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Alert people to evacuate the area.
Activate nearest fire alarm or call for emergency response.
Assess the situation: Only if it is safe to do so, try to extinguish the fire.
Close lab doors to confine fire.
Have a person who is knowledgeable about the lab and the incident
ready to assist emergency personnel.
While Working in the Lab:
Individual Protection

Shoes with full coverage
and good grip soles (no
sandals)

Restrain long hair, loose
clothing and jewelry

Wear a lab coat

Use appropriate eye, skin,
and hand protection

Wash hands often
Howard Hughes Medical Institute: Online Safety Course
http://www.practicingsafescience.org/
I'm here to help you load that TFA into the
centrifuge. Oops! I forgot my eye protection. I have
to go get it.
Well RUN for Pete's sake. Time is money!
Eye protection? You don't need any eye
protection.
Okay. Take your time and be aware of
your surroundings. It'd be ironic if you were
to cause an accident by trying to avoid an
accident, wouldn't it?
Think ahead and plan your moves. Be careful of your surroundings.
Eye Protection

Protects against risk of
flying objects or dust
particles, splashes of
hazardous materials or
harmful rays
Safety Glasses




Unbreakable lenses of
plastic or tempered
glass
For light-to-moderate
work
Can be prescription
lenses
Do not interfere with
contact lenses
Goggles


Work with significant
risk of splash of
chemicals or projectiles
Can be worn over
prescription glasses
Face Shield

Work with significant
risk of splash on face or
possible explosion
(including removing vials
from liquid nitrogen)

Face shield protects face
adequately but not eyes
Hand Protection






Protects against risk of cuts, abrasions, burns,
or exposure to hazardous materials.
Requires selection of the appropriate chemical
resistant gloves
Change gloves often – none are impermeable!
Wash your hands often to reduce contamination
of yourself and others.
Do not touch door knobs, telephones, etc. with
gloves – remove them.
If you must transfer your experiment to another
room



put all materials, including clean gloves, in a tray
with spill-proof edges
remove gloves and wash hands
Re-glove once you are in the alternate location
While Working in the Lab:


Follow universal
precautions
Handle unknowns as if
they were hazardous
While Working in the Lab:

Handle volatiles, aerosols
or fine powders in a
chemical fume hood
Contain
bioaerosols in a
biological safety
cabinet
Laboratory Chemical Hoods

The laboratory chemical hood is a
ventilated enclosure that protects you
from being exposed to chemical
fumes, gases, and aerosols that are
generated within the enclosure.
Protection is provided by room air that
is drawn into the hood and vented to
the atmosphere. The hood ventilation
provides further protection by diluting
the concentration of flammable gases
below explosion limits.

The sash must be lowered to the
operating position or vapours will spill
out.

The hood should always be on. Notify
the facility staff or the safety office
immediately if the hood is off or you
observe contaminants escaping from
it.
Using Laboratory Chemical Hoods

Plan your experiment. First, assess the risks of your
experiment. If a laboratory chemical hood is required, place
everything you need in the hood before starting.

Lower the sash. Always make sure the sash is pulled down to
the marked level. The best protection is provided when the
sash is brought to the lowest level for convenient operation.

Watch your activity. Use slow and deliberate motions. Place
supplies so that they do not obstruct the airflow at the airfoil
sill or the exhaust slots at the back of the hood. Do not work
within four inches of the airfoil sill.

Keep the work area uncluttered. Do not store chemicals in the
cabinet.
Storage of Hazardous Chemicals

Laboratory directors, managers and personnel are responsible
for understanding the general principles governing storage of
the various classes of chemicals and for complying with
McGill’s policy. This includes segregation of incompatible
chemicals, proper storage conditions (e.g., safety cabinets) and
limits on volumes of certain chemicals that may be stored in
the laboratory (e.g., flammable liquids).

See http://www.mcgill.ca/ehs/laboratory/labsafety/#3.6.3.1



Section 4. Handling and Storage of Laboratory Chemicals;
Section 7. Laboratory Ventilation And Fume Hoods
Appendix 1: Flammability Classification (NFPA) and Permissible
Container Sizes (OSHA)
Chemical Storage



Storage areas for corrosive, toxic, flammable,
and highly reactive chemicals should be near a
laboratory chemical hood.
Transfer hazardous chemicals from stock
containers to use containers only in a
laboratory chemical hood.
You should not store hazardous chemicals
inside the hood.
Rules of Thumb for Safe Storage
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Do not store more chemicals than you will need over a reasonable time
Always read the chemical's label and mark it with the date of receipt before
storing.
Never store highly reactive chemicals for longer than 6 months.
Never store a chemical with an obscured or missing label.
Separate chemicals into compatible groups and store alphabetically within
the groups (see section 4 of lab safety manual).
Designate separate storage areas for highly toxic chemicals.
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Flammables, organic acids and organic bases in separate trays in a flammables’
cabinet
Inorganic acids, strong bases in separate trays in the cabinet under the fume
hood
Toxic chemicals separately.
Oxidizers separately.
Corrosives separately.
Dry chemicals on shelves with raised edges, separated according to
incompatibility.
Flammables should never be stored in a frost-free refrigerator/freezer.
Class II Biosafety Cabinets

The type of hood present in most tissue
culture labs.

Protects you from being exposed to
infectious aerosols that may be generated
within the cabinet. You are protected by
room air that is drawn into the cabinet front
grill and the cabinet downward airflow. This
combination of airflows quickly prevents
respirable size particles from escaping into
your lab.

Protects your cultures from microbial
contaminants that are ubiquitous in room
air. Your cultures are protected by the
cabinet downward airflow, which is filtered
by high efficiency particulate air (HEPA)
filters. HEPA filters also treat cabinet
exhaust air.
Working in a Biosafety Cabinet

Use personal protective equipment


Wear a lab coat and gloves.
Plan your experiment in advance

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Place everything you need in the cabinet or on an adjacent
cart before starting the experiment.
Designate separate areas on the cabinet work surface for
clean materials and dirty materials.
Check that the vacuum bottle contains fresh disinfectant
(e.g., a one-in-ten dilution of household bleach) and that
there is enough space to collect all waste.
Do not use solvents in the presence of open flame.
Working in a Biosafety Cabinet

Use careful, sterile technique!
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Work with the sash lowered to the
indicated level.
Move slowly and deliberately with
planned movements.
Do not pass your hands over sterile
preparations or containers.
Discharge pipettes against the flask or
tube wall to avoid splashes.
Do not re-introduce pipettes into stock
containers – use another pipette.
Place disposable pipettes and dishes into
biohazard containers and non-disposables
into disinfectant.
Working in a Biosafety Cabinet

Wash hands before and after procedure. If gloves become
soiled, remove them, wash hands and put on new gloves.

Be careful of sharps!

Clean up at the end of the procedure and disinfect surfaces
with alcohol.

Before leaving the room, check everything! Then check
again!
How to Avoid Contaminating your Cultures,
You or your Coworkers

Cleanliness!!

You



Incubators

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


Keep free of clutter, particularly to maintain airflow
Keep surfaces clean and disinfected
Clean under the working surface regularly
The Room

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

Wash and disinfect thoroughly on a regular basis
Clean up spills immediately
Check for contaminated cultures regularly and remove immediately
Biosafety Cabinets


Wash your hands before and after any procedure
Avoid touching telephones, door knobs, equipment with gloved or dirty hands
Keep counters, floors and equipment clean and uncluttered
Do not overfill biohazard containers
Keep traffic to a minimum and have an organized schedule of use
Organization and Sterile Technique

Proper technique eliminates the need for routine use of antibiotics – save them for
when you really need them.
Using Autoclaves

Load the autoclave according to instructions. Do not use
without prior instruction.

Use loading racks/trays – never place any item directly on
the bottom or floor of the autoclave.

Starting the sterilization cycle. Make sure the door of the
autoclave is fully closed and the correct cycle has been
selected before starting the sterilization cycle.

Unloading the autoclave. Protect yourself from steam and
heat by wearing heat-resistant gloves. First, slightly crack open
the door. Wait a full five minutes if the autoclave load contains
just glassware, and no less than ten minutes when you are
autoclaving liquids. Then remove the load and let the
glassware stay on the racks 15 minutes before handling the
individual pieces.
Autoclaving Liquids

Loosen caps. Before loading containers of liquids into the
autoclave, the caps must be loosened to avoid having the
bottles shatter.

Guard against spills. Use a tray with a solid bottom and
walls to contain bottles and catch any spills. Add a quarter
to a half-inch of water so the bottles will heat more evenly.

Unload with extreme caution. Always wear a rubber
apron in addition to your standard protective equipment
when removing bottles that contain liquids from the
autoclave. Be alert for a bottle still bubbling - it could
explode easily if touched. Let the load stand in an out-ofthe-way place for a full hour before handling.
Before Leaving the Lab:
Turn off:
 Gas
 Water
 Power supplies
 Vacuum lines
 Compression lines
 Heating apparatus
Before Leaving the Lab:




Identify and package
waste, dispose properly
Put away experimental
apparatus, chemicals, etc.
Lock/out and tag/out
defective equipment
Decontaminate work
surfaces and equipment
Before Leaving the Lab:




Return borrowed
equipment, apparatus,
etc.
Leave lab coat in the lab
Wash
Close and lock door
Before Leaving the Lab:
 Clean
up your mess
 Leave
the lab safe for everyone

Perform a final check!!!
MODULE 2
Workplace Hazardous
Materials Information System
(WHMIS)
http://www.hc-sc.gc.ca/hecs-sesc/whmis/msds.htm
“No employer may allow the use, handling or
storage of a Controlled Product
in a workplace unless the product carries a label
and a material safety data sheet which meet the
requirements of this Act and the regulations
and unless the worker has received the training
and information required to carry out the work
entrusted to him safely”
- Article 62.1, An Act respecting occupational health
and safety R.S.Q., S-2.1
What are Controlled Products?
Learn the universal symbols for each class
Class A:
Compressed Gas
Class B:
Flammable and
Combustible
Material
Controlled Products
Class C:
Oxidizing Material
Class D Controlled Products
Division 1:
Materials Causing Immediate
and Serious Toxic Effects
Division 2:
Materials Causing Other Toxic
Effects
Division 3:
Biohazardous Infectious
Materials
Controlled Products
Class E:
Corrosive Material
Class F:
Dangerously Reactive Material
Not Classified as Controlled
Products:
explosives
radioactive
materials
Consult the McGill Laboratory Safety
Manual

Learn how to store each class of chemical and
separate incompatible classes of chemicals

Learn the limits on storage of hazardous
chemicals in the laboratory (e.g., flammable
liquids)
http://www.mcgill.ca/ehs/laboratory/labsafety/#
3.6.3.1
Class A: Compressed Gas
Characteristics
Gas inside cylinder is
under pressure
The cylinder may
explode if heated or
damaged
 Sudden release of high
pressure gas streams
may puncture skin and
cause fatal embolism
Class A: Compressed Gas
Precautions
Transport and handle
with care
Make sure cylinders are
properly secured
Store away from
sources of heat or fire
Use proper regulator
Class B: Flammable and Combustible
Material Characteristics
May burn or explode
when exposed to
heat, sparks or
flames
Flammable: burns
readily at room
temperature
Combustible: burns
when heated
Class B:
Flammable and Combustible Material
Precautions
Store away from
Class C (oxidizing
materials)
Store away from
sources of heat,
sparks and flame
Do not smoke near
these materials
Class C:
Oxidizing Material
Characteristics
Can cause other
materials to burn or
explode by providing
oxygen
May burn skin and
eyes on contact
Class C:
Oxidizing Material
Precautions
Store away from Class
B (flammable and
combustible) materials
Store away from
sources of heat and
ignition
Wear the
recommended protective
equipment and clothing
Class D, Division 1
Materials Causing Immediate and
Serious Toxic Effects
Characteristics
May cause immediate
death or serious injury if
inhaled, swallowed, or
absorbed through the skin
Class D, Division 1
Materials Causing Immediate and
Serious Toxic Effects
Precautions
Avoid inhaling gas or vapours
Avoid skin and eye contact
Wear the recommended
protective equipment and
clothing
Do not eat, drink or smoke
near these materials
Wash hands after handling
Follow procedures for disposal
of chemical waste
Class D, Division 2
Materials Causing Other Toxic
Effects
Characteristics
May cause death or permanent
injury following repeated or longterm exposure
May irritate eyes, skin and
breathing passages: may lead to
chronic lung problems and skin
sensitivity
May cause liver or kidney
damage, cancer, birth defects or
sterility
Class D, Division 2
Materials Causing Other Toxic
Effects
Precautions
Avoid inhaling gas or vapours
Avoid skin and eye contact
Wear the recommended
protective equipment and
clothing
Do not eat, drink or smoke
near these materials
Wash hands after handling
Follow procedures for
disposal of chemical waste
Class D, Division 3
Biohazardous Infectious Material
Characteristics
Contact with microbiological
agents (e.g., bacteria,
viruses, fungi and their
toxins) may cause illness or
death
Class D, Division 3
Biohazardous Infectious Material
Precautions
Wear the recommended
protective equipment and
clothing
Work with these materials
in designated areas
Disinfect area after
handling
Wash hands after
handling
Class E: Corrosive Material
Characteristics
Will burn eyes and skin
on contact
Will burn tissues of
respiratory tract if inhaled
7
Class E: Corrosive Material
Precautions
Store acids and bases in
separate areas
Avoid inhaling these
materials
Avoid contact with skin
and eyes
Wear the recommended
protective equipment and
clothing
Class F:
Dangerously Reactive Material
Characteristics
May be unstable,
reacting dangerously to
jarring, compression,
heat or exposure to light
May burn, explode or
produce dangerous
gases when mixed with
incompatible materials
Class F:
Dangerously Reactive Material
Characteristics
Store away from heat
Avoid shock and friction
Wear the recommended
protective equipment and
clothing
Other Important Universal Signs
What do these symbols mean?
Labels
WHMIS
Main Elements
- supplier
- workplace
- laboratory
Material safety data sheets
- supplier
- workplace
Training
- core
- job specific
DIMETHYL
MEGADEATH
Supplier’s Label
Learn how to read them
TOLUENE
PRODUCT
SULFONIC
IDENTIFIER
ACID
RISK PHRASES:
Highly irritating to skin, eyes, and nose
HEALTH HAZARD DATA:
Strong Acid: Treat as sulphuric acid
EFFECTS OF OVEREXPOSURE: ACUTE
PRECAUTIONARY MEASURES:
Supplier
identifier
ABC Chemicals
123 Chemical Drive
Chemical City
1-800-CHE-MICAL
PERSONAL PROTECTIVE EQUIPMENT
EYE: Face shield and goggles
GLOVES: Rubber
OTHER CLOTHING AND EQUIPMENT:
Rubber apron, rubber boots
FIRST AID:
EYES: Flush with water for 15 minutes. Consult
with physician
SKIN: Flush with water as per sulphuric acid
INGESTION: Treat as per sulphuric acid. Consult
with physician
Refer to Material Safety
Data Sheet
FRANCAIS AU VERSO
Workplace Labels Required:
On controlled products
produced and used in the
workplace,
 if the label becomes illegible.
received from a supplier and
transferred to another
container*
*not required if transferred material is
used in its entirety prior to the end of the
work shift
Workplace Labels
in Research Labs
 Product identifier (name)

MSDS must be available
in the lab
Agent
Green
Material Safety Data Sheets




Supplier must provide
Accessible to ALL
workers in the workplace
Must be kept up to date
Must be made available
to doctor in the event of
exposure
What’s in an MSDS?
Product Information
Item Name
Company's Name
Emergency Phone #
Part Number
MSDS Preparation
Ingredients
What’s in an MSDS?
Physical Properties
Appearance and Odor
Volatility
Solubility
pH
Fire and Explosion Data
Extinguishing Media
Flash point
Special Fire Fighting Procedures
Unusual Fire And Explosive Hazards
Definition: Flash Point

The flash point is the lowest
temperature at which a liquid
produces enough vapour to
ignite in the presence of a
source of ignition.

The lower the flash point, the
greater the risk of fire.
What’s in an MSDS?
Product Reactivity
Stability
Conditions To Avoid
Materials To Avoid
Hazardous Decomposition Products
Hazardous Polymerization?
Toxicological information
LD50-LC50
Route Of Entry
Signs/Symptoms Of Overexp
Carcinogenicity
Definition: Threshold Limit Value



TLV (TWA) is an 8-hour timeweighted average believed to be the
average concentration to which most
workers can be exposed during an 8hour workday, day after day, without
harmful effects.
TLV (STEL) is a 15 minute “short
term exposure limit”
Ceiling (C) is a maximum
concentration never to be exceeded
Daily exposure
limit
Definition: LC50

LC50 (Lethal Concentration50) is
the amount of a substance in
air that, when given by
inhalation over a specified
period of time, is expected to
cause the death in 50 per cent
of a defined animal population.
Definition: LD50

LD50 (Lethal Dose50) is the
amount of a substance that,
when administered by a defined
route of entry (e.g. oral or
dermal) over a specified period
of time, is expected to cause
the death of 50 per cent of a
defined animal population.
What’s in an MSDS?
Preventive measures
Respiratory Protection
Protective Gloves
Eye Protection
Other Protective Equipment
Ventilation
Storage
Spills
First Aid Measures
Which one of the following two procedures presents
the greater risk of producing a harmful effect?
Procedure 1: A technician handles 0.26 ml of ethyl methanesulfonate (EMS)
in preparing 100 ml of EMS sucrose solution needed for mutagenesis
studies with flies. The technician performs this procedure once every three
months. EMS is a possible human carcinogen. The oral LD50 for EMS is
470 mg/kg in mice, a moderately toxic rating.
Procedure 2: A technician prepares agarose gels daily. The concentration of
ethidium bromide in the gel box is 5 µg/ml. Ethidium bromide is a powerful
mutagen though its effects on humans are unknown. The reported
subcutaneous LD50 in mice in one study was 100 mg/kg, a moderately toxic
rating.
 Procedure 1 presents the greater risk of producing a harmful effect.
 Procedure 2 presents the greater risk of producing a harmful effect.
 The risks associated with procedures 1 and 2 are about the same.
Answer on next slide:

Answer 2 presents the greater risk because the
agent is used daily rather than every three
months.
Controlled Products Characteristics and Precautions
NFPA labels (National Fire
Protection Association)

Also valuable is the National Fire Protection Association's
labeling system that shows the type and the degree of a
chemical hazard.

The labels are commonly included on chemical containers.

The labels are diamond-shaped and color-coded.





Blue indicates the health hazard.
Red indicates the fire hazard.
Yellow indicates the reactivity hazard.
White gives special information such as water or oxidizer
incompatibility.
In each field, the degree of the hazard is rated from 0 to 4,
with 4 being the greatest hazard and 0 indicating no significant
hazard.
Risk Assessment
Any experiment involving a toxic chemical requires
planning to help you determine the potential risks and to
aid in the selection of safe practices.
Identify chemicals to be used.
Identify circumstances of use.
Consult sources of information.
Evaluate chemical toxicity.
Consider possible routes of exposure.
Select appropriate safety practices.
Prepare for contingencies.
Final Words
Safety is YOUR RESPONSIBILITY
Get the proper training
If you don’t know or are unsure, then ASK