Laboratory safety training

1. Developed for Science Laboratories
September 2002

2. This training presentation has been created
for those who are required to undergo
general laboratory safety training and have
not taken this training with their supervisor.
It is also recommended for those laboratories
that do not fall under regulation, but are
needing a safety laboratory overview.
There are hyperlinks throughout the presentation. Where there is the following
icon additional training is required. Training

3. Introduction
Laboratory safety is the key to reducing injury and
illness. There are many exposures in the laboratory
that pose a hazard to your health and you may have
never considered them as a hazard before. It is
important to have proper training so you, as the
employee, are aware of the potential dangers that
may threaten your health or life.

4. Introduction
As you go through this training module, you will have
a better understanding regarding the concept of
safety and how safety is utilized in every aspect of
your life. The University has an obligation to protect
each student, assistant, faculty and staff. There are
also regulations that pertain to ISU to ensure
compliance. Lets take a look at the different agencies
that are involved!

5. Agency Information
Several agencies are involved in safety compliance at
the University. The following agencies impact our
laboratories as far as safety is involved.
National Institute of Health (NIH)
In charge of laboratories that use or contain
recombinant DNA. They specify the practices for
constructing and handling the rDNA.

6. Centers for Disease Control and Prevention (CDC)
In charge of laboratories that use or contain infectious
agents. They are focused on protecting personnel and the
laboratory environmental from exposure to infectious
agents. They are also aiming at preventative measures by
adhering to strict containment.
Agency Information

7. Agency Information
Occupational Safety and Health Administration
(OSHA)
OSHA primarily monitors Hazardous Materials, Hazard
Communication, Bloodborne Pathogens, and
Occupational Exposure to Hazardous Chemicals in
Laboratories at ISU. OSHA is in charge of monitoring
work conditions and eliminating physical and health
hazards at the work place.

8. Environmental Protection Agency (EPA)
Agency Information
The EPA is in charge of the wastes that ISU generates
and stores. Many hazardous wastes are used in
research laboratories and they pose a threat to
humans, animals, plants, and the environment. There
are also biohazardous wastes that are regulated and
must be properly decontaminated and disposed of.

9. Agency Information
Environmental Health & Safety Office (EHS)
The EHS office plays a large role in overseeing the
various activities that take place on campus. They
interpret laws and regulations, and maintain up-to-
date records of current health and safety standards.
The duties range from fire safety and laboratory safety
to training and emergency response.

10. Why Training?
Training is by far the most important aspect
of safety. Training not only reduces
illness/injury but also increases worker
efficiency and awareness. Training is
required by regulation and ISU has high
regard for fulfilling this commitment.
Preventing hazards and increasing awareness
is the goal for the EHS office.

11. Who is in Charge of Training?
Everyone on campus is in charge of training. The
individual is responsible for participation of the
training, comprehending the information, and
utilize the information for the specific duties. The
supervisor is in charge of providing proper and
effective training. The EHS office provides
assistance to departments in achieving regulatory
compliance and developing proactive strategies.

12. Introduction to the Laboratory

13. Let’s Get Started!
Working in a laboratory can be an
exciting experience. It can also pose
many threats and hazards that a
traditional classroom does not. That
is why it is important to know your
surroundings. Know where the exits
to your room are. There may be
more than one exit which could be
critical in the case of an emergency.
Your supervisor will go over the
emergency action plan including the
escape route procedures for your
room.

14. Know Your Surroundings
It is also recommended to be aware of the fire
extinguishers in location to your laboratory. In order
to fight a fire one must undergo the proper training.
Campus personnel are highly encouraged to not fight
fires. In the event of a fire, the first response is to
evacuate the area and notify the fire department!
Any campus phone will initially direct calls to the ISU
Police Dept. and from there the fire dept. will be
dispatched. From any campus phone just dial 911.
Do not wait any longer than necessary to call, time is
of essence!

15. Know Your Surroundings
Know where the fire alarm is in proximity to your
laboratory. Is it right down the hall or is it in the
stairwell? If there is a fire, a quick response is
the best response. Have your supervisor show
you the closest alarm. The fire safety training will
detail the safety procedures for your lab.
Training

16. Many laboratories contain hazardous substances.
A hazardous substance is defined as a
material/substance that poses a physical or health
hazard. This includes both chemicals and
biological agents.
A Biohazard is defined as any organism that is
capable of replication and is capable of causing
disease in human, animal or plant.
There are differences between a physical hazard
and a health hazard. Let’s take a look.
Know Your Surroundings

17. A health hazard has the following
characteristics:
Know Your Surroundings
Carcinogen
Toxic or highly toxic
Reproductive Toxins
Irritants
Corrosives
Sensitizers
Hepatotoxins
Nephrotoxins
Neurotoxins

18. A physical hazard has the following
characteristics:
Know Your Surroundings
Explosive
Flammable
Oxidizer
Pyrophoric
Organic peroxide
Compressed gas
Combustible liquid
Unstable (Reactive)
Water-reactive

19. When physical hazards and health hazards
exist, it is very important to know where
the eye wash/safety shower is located.
Unexpected accidents do occur and
knowing where to go at the time of an
emergency can reduce injury/illness.
Know Your Surroundings

20. First aid kits have a variety of quick relief
items. If your lab has a first aid kit, find out
where it is. If more than first aid is needed,
it is recommended to go to Student Health
Services for further treatment. In an event
that would require more than first aid to be
treated, report it to the EHS office within
the next 24 hours.
Know Your Surroundings

21. When there are chemical, biological, or
radioactive agents being used, an
emergency spill kit should be available. If
there is a spill kit in your lab, find its
location. Further Spill information will be
addressed later in this presentation.
Know Your Surroundings

22. Each laboratory has a telephone in a
designated area for use. The emergency
contact numbers are posted near the phone
in every laboratory on ISU campus.
Know Your Surroundings

23. Know What Hazards are Present

24. Each lab is faced with different hazards.
There could be exposure to biological,
chemical, or radioactive material, which
may pose a variety of physical and/or
health hazards. This training will be
primarily focusing on biological hazards
and some chemical hazards. Let’s look at
these hazards a little closer.
Hazards in your Lab

25. An infectious agent is an organism capable of
producing infection or disease in human, animal or
plant.
Recombinant DNA (rDNA) is defined as molecules
that are constructed outside living cells by joining
natural or synthetic DNA segments to DNA
molecules that can replicate in a living cell or
molecules that result from the replication of those
described above.
Hazards in your Lab
A biological hazard includes an organism or material
of biological origin that could potentially cause harm
to humans, animals, or plants.

26. Infectious agents pose a threat because these
agents can cause illness or death to both people
and animals. Special precautions must be taken to
reduce the potential release of these agents. Each
laboratory that is using an infectious agent must
perform an additional lab specific training. This is
to communicate the hazards of the agent that is
specific to your laboratory.
Hazards in your Lab
Training

27. Infectious agents fall under CDC guidelines.
Depending on the organism you are working
with or exposed to will depend on the proper
safety precautions to take. These are
designated at different Biosafety Levels
(BSL). The more pathogenic the agent, the
more stringent the safety precautions are.
Hazards in your Lab

28. The lab that works with infectious agents,
depending on the Biosafety Level (BSL), will
vary in accordance to the safety equipment
that is used, the facility (lab) design, the
equipment that is used, and the practices
that must be followed.
Hazards in your Lab

29. Biosafety levels will be
discussed in detail by
your supervisor during
the Lab Specific
Training.
Hazards in your Lab
Training

30. rDNA can also poses a threat to human and
animal health. Depending on the molecules that
are being manipulated, the hazards will vary.
rDNA can be manipulated using cultures, animals,
plants, and also humans.
The risk will depend on several factors including
the organism and the media being used. The
containment levels will depend on the organisms
that carry the recombinant DNA.
Hazards in your Lab

31. The term containment in the laboratory setting is
defined as the confinement of a biohazardous agent
that is being cultured, stored, manipulated,
transported, or destroyed in order to prevent or
limit its contact with people and/or the
environment. Methods used for containment
include physical and biological barriers and
inactivation using physical or chemical means.
Containment is integrated into the distinguished
biosafety levels.
Hazards in your Lab

32. Chemicals can pose a significant hazard. They
should be limited to the use under a properly
working fume hood. Chemicals can release
hazardous fumes which not only harm the
environment, but they can be a major health threat.
They must be handled carefully and disposed of
properly.
Hazards in your Lab

33. When a chemical is in the laboratory, the hazards of
that chemical must be communicated to you.
According to Occupational Safety and Health
Administration (OSHA), a Chemical Hygiene Plan
(CHP) is required to relay information regarding
procedures, equipment, PPE, and work practices
that are capable of protecting employees from
health hazards.
Your supervisor is in charge of providing the
information contained in the CHP to you.
Hazards in your Lab

34. Lab Safety
The following guidelines have been established to
minimize the hazards in a laboratory setting. It is
important to take responsibility for your actions
and to keep in mind that irresponsible acts could
have lasting future effects.
The next portion of the training is simply review
and is meant to remind you of responsible
conduct in a laboratory setting. As the hazards
increase, the risks increase, and the responsibility
must increase.

35. Lab Attire
 No open-toed shoes
 No shorts unless a lab coat is used
 Restrain hair when working with hazardous
materials
 Remove protective clothing in public
 Use the proper Personal Protective Equipment for
the job
You should remember the following:

36. Personal Habits
 Do not eat, drink, smoke, chew gum or
apply cosmetics, or remove/insert contact
lenses while in the laboratory
 Do not store food or beverages in the lab or
in chemical refrigerator
 Do not mouth pipette
 Wash hands before leaving laboratory or
after handling contaminated material
Personal habits play a large role in minimizing
hazards. The following measures must be
taken:

37. Safe Practices
These safe practices should be followed to ensure
safe working conditions:
 Do not use chipped or cracked glassware
 When working with hazardous materials, have a
second person nearby
 Know emergency procedures
 Keep the laboratory neat and clean
 Use hazardous chemicals under a fume hood and
biohazardous materials under a biosafety cabinet
(BSC)
 Decontaminate as needed
 All procedures should be performed to minimize
aerosol

38. Know details/location of your vital
Laboratory Information!

39. Each lab contains an Emergency Action Plan which is
required for emergency situations. This is used to
inform faculty/staff/students of the procedures to
follow in the event of an emergency. This includes
the following information:
Laboratory Information

40. The labs that are using infectious agents and/or rDNA
are also required to have the ISU Biosafety Manual
available. Specific guidelines for research are
outlined in this manual. If any questions arise
regarding regulations, this manual should be used as
a resource.
Laboratory Information

41. The Chemical Hygiene Plan (CHP) is similar to the Hazard
Communication Program, but this is specifically for laboratories
with chemicals. This is to relay information regarding
procedures, equipment, PPE, and work practices that are
capable of protecting employees from health hazards. If you
are working with acids, it would be nice to know what personal
protective equipment is necessary and what health hazards are
associated with it.
Laboratory Information

42. Chemical Hygiene
Methods and observations that may be used to detect
the presence or release of a hazardous chemicals.
Physical and health hazards
Measures employees can take to protect themselves
from hazards
Details of the CHP
According to the laboratory standard, for those labs that
have chemicals the following training topics must be
addressed:
This training is covered in the Hazard Communication Training
Presentation
Training

43. Methods and observations used to detect the
presence or release of chemicals will be
specific to your lab. A good indication of the
presence of a chemical is to rely on your
senses. Can you see it or smell it? Is a
monitoring device needed to detect it? This
can be important information if the chemical
is an asphyxiant and pushes oxygen out of
the room you are in.
Chemical Hygiene Plan

44. Chemical Hygiene
Protection against chemicals is a combination of
work procedures or practices, emergency
procedures, and PPE. Some chemicals can only
be handled under certain conditions. It is
important to use proper handling procedures and
practices as advised. The emergency procedures
for chemical accidents is to first evacuate the area
and then notify your supervisor, ISU campus
police and EHS office if necessary.

45. Chemical Hygiene
Each lab that has chemicals will have a lab
specific Chemical Hygiene Plan. This plan
will detail the procedures and practices for
your specific location. It is your
responsibility to make sure that you review
the CHP.

46. It is important to know as much about a
chemical as possible. The most dangerous
substance is the one that has no label.
Communicating information is essential in
the science field.
Labeling

47. Labeling is also required for chemicals that
are in a secondary container unless it is to
be used within an 8 hr period. When
labeling the secondary container the
following information must be included:
Labeling
Identity of the hazardous chemical
Appropriate hazard warnings
Name and address of the chemical
manufacturer, importer, or other responsible
party.

48. There are several constituents that are
covered in the Laboratory Standard. Among
these constituents includes labeling and the
MSDS. Let’s look more closely at the content
of the MSDS.
MSDS
MSDS

49. MSDS
In addition to labeling in a laboratory, the
next most important type of communication
regarding hazards is the MSDS. This is the
acronym for Material Safety Data Sheet.
This will communicate the information
necessary regarding hazards associated
with chemicals and also biological agents.

50. So what is an MSDS? An MSDS is a document
that relays vital information about certain
chemicals and biological agents. Check below
to see what content the MSDS contains!
MSDS
MSDS Content
Click here!

51. Reading the MSDS
Each component of the MSDS is broken
down into sections. Each section goes into
detail about what precautions to take and
the characteristics of the substance. This is
for both chemicals and biological agents.
See the below links for example MSDSs.
MSDS Chemical
MSDS Biological Agent

52. Why is an MSDS important? When you know
characteristics about a substance it can aid in
precautionary measure to take when using it.
Also, if there is a spill either on a surface or on
your skin, the MSDS can supply you with the
information needed for first aid.
Laboratory Information

53. MSDS
The MSDS to every chemical in your lab
must be available to you. It may be in
a notebook in your lab. Make sure you
find the location of the MSDSs in your
room.
MSDS

54. Hazard Communication
If chemicals are used, Hazard Communication
training must be conducted. This training is
specifically for the labs that use chemicals and
the training can be accessed through the EHS
website.
Training

55. The Exposure Control Plan is required for the labs that
have exposure to human blood or Other Potentially
Infectious Material (OPIM). This includes human
blood or human blood products such as semen or
vaginal secretions. This also includes fluids such as:
cerebrospinal, synovial, pleural, pericardial, peritoneal
and amniotic. Saliva and body fluid that is visibly
contaminated with blood also is considered OPIM.
Exposure Control Plan

56. Exposure Control Plan
The Exposure Control Plan is established to make
an exposure determination and a schedule of
implementation for methods of compliance, HBV
vaccination and post-exposure evaluation including
follow-up, HazCom, and recordkeeping. If you are
working with human blood or OPIM, you must talk
with your supervisor and locate the Exposure
Control Plan.
There would only be a select few laboratories that
this standard would apply to.

57. Personal Protective Equipment

58. What is PPE?
PPE is short for personal protective equipment. This is the
equipment that is necessary to protect yourself from
hazardous and biohazardous materials. PPE could be gloves,
safety glasses, lab coat, shoe covers, respirator or any other
item that could protect you from dangerous materials that you
may encounter in the lab.

59. Knowing what to use and when to use it is the key to properly
protecting yourself. There could be situations that would be
more of a risk and require more PPE than others. The next few
slides will help you in determining what you should be using!
When to use PPE

60. Chemical usage poses a variety of hazards.
They can be flammable, corrosive, even toxic
just to name a few. Taking all precautions
to avoid physical and/or health problems is
the number one goal. You can never be too
cautious!
When to use PPE

61. Chemicals & PPE
When chemicals are being used there is
always the possibility of splashing. The proper
PPE to use when chemicals are involved would
include:
Safety glasses
Gloves
Respirator (depending on the chemical and the exposure
duration)
The Permissible Exposure Limit (PEL) for an 8 hr. shift can be
obtained from the PEL/BEL reference guide. Contact EHS if
questions arise.

62. Biological agents as well as rDNA can pose serious
health problems. Pathogens can cause disease and
possible death in animals and humans. Containment
of the agent/material and proper handling
techniques are the best methods for minimizing risks
that are associated with biological hazards. rDNA
can also be a threat depending on the host and the
vector that is being used.
PPE is only one way that you can protect yourself. It
is your responsibility to ensure that the proper PPE is
used.
When to use PPE

63. When biological agents and/or rDNA are
being used the following PPE must be used:
Biological Agents & rDNA
PPE
Safety glasses
Gloves
Lab coat
Respirator (Depending on the agent and whether or not an
aerosol is created)

64. Animals have the ability to bite and scratch
unlike many of the other hazards. This can be of
concern when the animal is infected and can
transmit disease to a human. This is called
zoonotic. To avoid infection, proper handling
procedures must be utilized. The appropriate
handling in combination with the PPE can help to
reduce risk.
When to use PPE
www.animalpicturesarchive.com

65. Animals & PPE
When animals are being used the following
PPE must be used:
Gloves-May need a variety to prevent animal bites.
Lab coat-Needed to protect clothing and used as a barrier. To
prevent the pathogen from leaving the lab, the lab coat must be
worn and must not leave the lab.
Safety glasses-Must be worn to prevent possible injury to the
eyes by scratching or flying saw dust bedding. Splashing or
aerosol also requires glasses.
Respirator-May be needed depending on the pathogen. Check
with safety office.
Shoe covers-Used in the animal room and labs that work with
biological agents.

66. Arthropods are associated with potential risks when
they escape since some are vectors of infectious
human diseases. Precautions need to be taken to
minimize the hazards associated with arthropods.
Communication during lab specific training can help
you understand the potential risks associated with the
arthropod that you come in contact with.
When to use PPE

67. Arthropods & PPE
Depending on the arthropod that you
encounter in the lab, the PPE could vary. In
most cases the PPE below should be used
used.
Gloves
Safety glasses (Use if arthropod is infected or if there is a
chance of the arthropod spraying or jumping in your eye)
Lab coat (If arthropod is known to be infected)

68. Proper Use of Personal
Protective Equipment (PPE)

69. Use of PPE
Knowing how to properly use PPE can be
the key to adequate protection. Not only
do you want to make sure it is the proper
size for you, but also make sure you are
wearing it properly. If it is too big or too
small, it is not right for you! Let your
supervisor know if you need a different
size.

70. Use of PPE
The main piece of equipment that must be used
appropriately is the respirator. If you work with
infectious agents, a respirator may be required
depending on the biosafety level (BSL) of the agent.
The more pathogenic the agent, the more PPE that is
required. If a respirator is required, you will be part
of the ISU Respiratory Protection Program. This
requires a physical, fit-test, and additional training
before use. If the respirator is not properly fit to you,
serious health problems can result.
Training

71. If you think you need a respirator talk
with your supervisor or contact EHS
(438-8325)
Use of PPE

72. Location & Availability of PPE
Each Lab should have a designated area for the PPE
that is used. The PPE should also be readily
available when working with materials that require
it. If the proper equipment is not available, you
should not proceed with the work. Notify your
supervisor when you need to order PPE. Give an
advanced notice when possible due to shipping
time. Always consider in advance what you will
need when conducting research.
PPE

73. Maintenance of PPE
In order to be able to rely on your equipment, you
must take care of it. Keep the PPE clean and
disinfected. It should be cleaned before and after use
when possible. Proper maintenance can not only
extend the life of the equipment, but also keep cost
down. Keeping the cost down on the Personal
Protective Equipment can help to better allocate the
money toward larger equipment needed for research.

74. What is a Sharp?

75. What is a “Sharp”?
A sharp is defined as any instrument, tool, or
item that has rigid, acute edges,
protuberances or corners capable of cutting,
piercing, ripping or puncturing such as
syringes, blades, and broken glass. Items
that have the potential for shattering or
breaking are also considered sharps.
http://www.ehrs.upenn.edu/training/bloodborne/bloodborne.html

76. Safe Practices
When using a sharp there is a risk of being cut
by the object and possible infection occurring
depending on what the sharp was used for. If
hypodermic needles are used, special
precautions must be taken to reduce the risk of
a needlestick. After use of the needle do not
recap, place directly in the sharp container.

77. Disposal of Sharps
All sharps must be placed into a rigid,
puncture and leak-resistant container that
is also impervious to moisture. The
sharps container must be labeled either
with “Biohazard” or “Infectious Waste”.
Do not over fill the sharps container.
http://www.ehrs.upenn.edu/training/bloodborne/bloodborne.html

78. Disposal of Sharps
When the sharps
container is full it must
be collected by the EHS
office. A waste pick-up
form can be completed
and a collection can be
scheduled. A waste
pick-up form can be
obtained from the EHS
office (438-8325).

79. Sharps Log
If an injury occurs as a result of a sharps
accident, it must be recorded in the Sharps Log.
This log is kept by the supervisor and is viewed
upon annual inspection. The main purpose of
the log is for risk assessment and
documentation. If a sharps injury involves an
infectious agent or anything beyond first aid, it
is advised to seek medical attention at the
Student Health Services.

80. What is a Risk Group (RG)?

81. Risk Group
According to National Institute of
Health, there is a risk associated
to certain research. This is
targeting the laboratories that
work with infectious agents and
rDNA. A risk group (RG) is
determined by the nature of the
research. The more hazards
there are with the agent/material,
the higher the risk group and
therefore more stringent the
containment and work practices
become.
RG 1
RG 2
RG 3
RG 4
The Risk

82. Risk Groups
The risk groups are broken down by the following
Agents are not associated with disease in healthy adult humans
(low individual & community risk) Example: Bacillus subtilis
Agents are associated with human disease, which is rarely
serious, and for which preventive or therapeutic interventions
are often available (moderate individual risk, limited community
risk) Example: Streptococcus aureus
Risk Group 2 (RG2)
Risk Group 1 (RG1)

83. Risk Group 3 (RG3)
Risk Groups
Agents are likely to cause serious or lethal human disease for
which preventive or therapeutic interventions are not usually
available (high individual risk & high community risk) Example:
Ebola Virus
Agents are associated with serious or lethal human disease for
which preventive or therapeutic interventions may be available
(high individual risk but low community risk) Example: Hantavirus
Risk Group 4 (RG4)

84. Risk Groups
The RG in your lab is to be determined
by your supervisor. The information is
to be communicated to you during your
lab specific training.
Training

85. What is a Biological Safety Level
(BSL)?

86. Biological Safety Level
Along with the Risk Group there is also an
associated Biological Safety Level (BSL). This has
the same principle as the RG in that as the risk
increases, the containment and practices become
more stringent. For most cases, if your lab is a RG
1 then the BSL is 1.
CDC and NIH have set guidelines to help protect
individuals who work with infectious agents and/or
rDNA.

87. Biosafety Levels (BSL)
The different BSLs have
been established as
preventative measures
against human infection.
Taking precautionary
measures can help to
minimize hazards
associated with infectious
agents.

88. BSL
As the risk increases, the BSL also
increases. Each BSL has specific
containment practices that CDC and NIH
have advised. When the lab specific
training is conducted by your supervisor,
she/he will go through the BSL guidelines
for your lab.
Training

89. Signs and Labeling

90. Labeling
Biological material that others may have
access to should be labeled. This would
include items such as cultures. The label
should describe what the content is and also
any precautions. An example is below.
Escherichia coli
Precautions:
Must use Eye protection, lab coat, gloves
Only use under BSC

91. Labeling
Chemical labeling has been briefly touched on earlier
in the presentation. One must remember that if any
chemical is transferred to a secondary container, this
container must be labeled. If the chemical will be
used by the end of the work shift, then labeling is
not necessary. Good science practices would
encourage you to label all containers.
So what is included on the Label?
Click here

92. Signs
Signs are a way of communicating important
information. It is a way to heighten
awareness about hazards that exist.
There may be signs on laboratory
refrigerators reminding you that no food or
drink can be stored in it. There may be
radioactive or biological materials that could
possibly be absorbed in food.
There may also be signs which denote that
PPE must be used.

93. Signs
Labs which use rDNA and
infectious agents must have a
sign posted on the outside of the
door. Before someone enters the
lab, they will have the information
they need to protect themselves.
Always read the signs carefully so
you know what precautions to
take.

94. Containment of Laboratory

95. Containment
Containment of a laboratory is to confine
organisms containing rDNA molecules and/or
infectious materials. The purpose is to
reduce or eliminate the potential for
exposure of the laboratory worker, persons
outside of the laboratory, and the
environment from organisms containing
rDNA and infectious materials.

96. Containment
Laboratory Practices
Containment Equipment
Special laboratory design
Let’s take a closer look
There are three elements to containment:

97. Practices and Techniques
Responsible practices and techniques are
required when working with hazardous
materials. This is one very important way to
reduce accidents/injuries. The following
practices should be followed in addition to
reviewing the Biosafety Manual:
Practices and Techniques

98. Safety Equipment
The Primary Barriers

99. Safety Equipment
Certain equipment is necessary to achieve
compliance and most importantly to provide
adequate protection.
The safety equipment that is needed is
known as primary and secondary barriers.
Let’s take a look at the difference in the
barriers.

100. Primary Barriers
Primary barriers are referring to protective
measures including engineering controls. This
includes not only PPE that has already been
covered, but it also includes safety cabinets, fume
hoods, vaccines and autoclaves.
It is important to know when this equipment
is to be used and how to properly use it.

101. Primary Barriers
Let’s look at the following barriers:
(Besides PPE)
Fume hood
Biological Safety Cabinet (BSC)
Autoclave

102. Fume Hood
The fume hood is used with chemicals. The
main function is to exhaust the vapors and
gases that are generated in the hood to the
outside. The hood is designed to minimize
your exposure to airborne contaminants. This
is not to be used with biohazardous materials.

103. Fume Hood Use
When using the fume hood you first need to make
sure the exhaust blower is operating and air is
entering the hood.
Remember, do not put your face inside the hood!
Minimize storage of chemicals in the hood
Clean spills immediately
Work with the sash at the proper operating level
as indicated by the arrows

104. Biological Safety Cabinet
The biological safety cabinet (BSC) is used as
a containment for infectious agents. The BSC
has a HEPA filter in the exhaust system to
protect the environment and yourself.
The (HEPA) filter is a high efficiency
particulate air filter. It is able to remove
particles at a size of 0.3 m with an efficiency
of 99.97%. It is also able to remove both
smaller and larger particles.

105. BSC
There are 3 classes of BSC that are used.
The higher the risk group and biosafety
level, the higher the class of cabinet that is
used.
If there is an infectious agent being used,
whether it is used in research animals or
cultured, it must be manipulated inside the
BSC.

106. BSC
When using this containment device, remember
to also use the proper personal protective
equipment. The following PPE should be
considered depending on the BSL that is required
for the organism that you use.
Gloves
Lab Coat
Shoe Covers
Safety Glasses
Respirator

107. Use of BSC
Always make sure that the BSC has been
decontaminated both before and after use.
Decontamination methods vary depending
on the infectious agent being used. Once
the decontamination of the cabinet is
complete, place the waste in a biohazard
bag for autoclave.

108. Use of BSC
Do not bring contaminated materials out of
the cabinet until they have been surface
decontaminated. If you are unable to
decontaminate, place the material into a
closed container to transfer it to the autoclave

109. Work Practices for the BSC
Remember to follow the work practices:
Work in such a way that your face is above the
front opening
Wait for 1 minute after placing hands/arms inside
the cabinet to stabilize the air flow
Work at least 4 inches from the inside edge of the
front of the grille

110. Remember to place all materials as far
back in the cabinet as possible
Limit the storage within the cabinet
Work Practices for the BSC

111. Autoclave
An autoclave is used to treat infectious
material and rDNA. As a standard for the
University, all material of this nature must be
autoclaved as a safety precaution. The
autoclave is able to render the material as
non-infectious.

112. The autoclave is able to reach a high
temperature to sterilize the agent. It is
important to know the standard operating
procedures (SOP) for the autoclave. The
SOP is located next to each autoclave. If the
temperature or pressure is inadequate, the
bag is overfilled, or the peak time is not long
enough the material will not be properly
decontaminated.
Autoclave

113. Autoclave
The autoclave is sensitive to
time, temperature, pressure,
water content, type of
container being used and the
type of waste being sterilized.
When using the autoclave there
are a few important point to
remember in order for the
process to be effective.

114. Procedures
The material being autoclaved must be placed inside
an autoclave red bag or in a pan (Do not overfill the
bag).
Add ~250-500ml water to the bag or pan for steam to
generate and properly circulate
A spore strip or other approved tape must be placed
on the outside of the bag or pan to verify that the
heat treatment was successful. You will have to verify
that a color change took place.

115. Procedures
The autoclave must reach a temperature of 121°C
(250 °F) for at least ½ hour at a pressure of 15psi
After successfully being autoclaved, the material can
be placed in regular trash.
Sharps must be incinerated. When the container is
full, a pick up can be scheduled through EHS and
from there Student Health Services will incinerate the
sharps container for proper disposal

116. Autoclave Log
Each time the autoclave is used, the log must
be completed by the individual using it. The
log is to be located at the site of the autoclave.
The information contained within the log is as
follows: Date
Generator Name (P.I.)
Location
Time the autoclave reached 121°C
Max Temp.
Pressure
Type of waste
If spores or tape were used
Weight
Person using autoclave

117. Autoclave Log
The log is required by EPA and it must be
maintained for 3 years by the department.
The purpose of the log is to ensure that
proper decontamination is taking place
If a new log is needed at the autoclave,
contact biology department or EHS.

118. Facility Design
The Secondary Barriers

119. Secondary Barriers
Secondary barriers are the facility design
and construction. These barriers are to
provide protection for the individuals
outside the lab, the community, and the
environment.

120. Secondary Barriers
An example of a Secondary barrier would be a
laboratory. It is separate from the classroom and
has limited access.
The animal room has limited access and is
only available to authorized personnel.
The autoclave is set away from the normal
traffic.
The handwashing facilities are located within
the lab and not accessible by the outside
students.

121. Decontamination

122. Decontamination
Decontamination is the removal or
neutralization of toxic agents or the use of
physical or chemical means to remove,
inactivate, or destroy living organisms. This
includes both sterilization and disinfection.

123. Decontamination
Decontamination is the responsibility of all
laboratory workers. Failure to
decontaminate can result in exposure to
infectious agents which can cause great
illness. Most decontamination can be
done by chemicals. This technique is used
only when autoclaving is not possible.
Continue on to see what would be best for
your lab.

124. Chemicals
There are a variety of chemicals that can
be used as an effective method of
decontamination. Depending on the
agent being used, the method to use may
vary along with the contact time. For
most organisms, a 1:100 chlorine solution
for 10-30 minutes is adequate. The
Biosafety Manual has a list of sterilizers/
disinfectants that can be used.

125. Autoclave
The autoclave is the most effective method to
use for decontamination purposes. As a
general rule of thumb, autoclave all materials
that are considered infectious agent, rDNA, or
resemble components of this nature. When in
doubt, AUTOCLAVE! If a material is not
capable of autoclave because of its size,
material, or it is stationary, then rely on
chemical disinfectant as a second option.

126. Incineration
There are other methods that can be used for
decontamination such as incineration. This is
the most reliable method, but it will destroy all
materials placed in it and it creates
environmental pollutants. If you must
incinerate, contact EHS.

127. Spills and Accidents

128. SOP
Spills and accidents can pose a serious
health and safety threat. When a spill
occurs, an aerosol can be created which can
make the material several times more
potent. The best measure to take in order
to protect yourself is to be prepared. There
should be standard operating procedures for
this type of situation in your lab.

129. What to Do?
Being able to recognize the hazards, mitigate the
spill, and notifying response authorities can be your
best defense. The first response to a spill should be
to evacuate the immediate area until the scope of
the hazard has been addressed. Seek medical
attention if necessary. Allow sufficient time for the
aerosol to settle before considering entering the
room. If you are responsible for clean up, proper
training shall be addressed.

130. How to Report
When a spill occurs, it must be recorded in the
Spill Log. The main purpose of the log is to
track an exposure in case of future
illness/injury. Report to your supervisor all
spills. If medical attention is needed, it is
suggested to go to Student Health Services.

131. Waste Management

132. Waste
Hazardous and biohazardous waste has
special guidelines for proper disposal. It is
important to properly dispose of waste to
ensure human and environmental health.
EPA regulates the waste that is generated
at ISU.

133. Waste
Waste can be classified as either hazardous or
biohazardous. Let’s take a closer look at the
differences.
Hazardous Waste- This is a waste which contains
the characteristics of being any of the following:
Toxic
Corrosive
Ignitable
Flammable
Oxidizer

134. Waste
A biohazardous waste is any waste that is
considered infectious and/or because of its
biological nature it can cause physical or
health hazards in humans, animals, plants or
the environment. This includes recombinant
DNA and other genetically altered organisms
and agents.

135. Proper Disposal
Waste that is considered biohazardous can be
disposed of in regular trash once it has been
rendered non-infectious. If a biohazard
labeled bag is used, make sure it is either
placed in a secondary bag or a completely
new bag that is not red.
http://www.lbl.gov/ehs/Medical/html/biohazardous.htm

136. Proper Disposal
Hazardous waste must be disposed of
through the Environmental Health & Safety
office. If you need to have hazardous waste
picked up, contact EHS for a pick-up form or
go online at www.ehs.illinoisstate.edu/
EHS 438-8325

137. You have now completed the General Lab
Safety Training presentation. If you have
any questions please take the time to ask
your supervisor or call EHS. By signing
the training form, you are stating that you
have completed the training and that you
understand the content within the
training. Please fill out the Certificate
of Completion on the next slide &
turn into your immediate supervisor.
Laboratory Safety Training
Certificate