LABORATORY ACTIVITY 1 Flashcards
(?) that result in injury, property damage, and even death are more common than we might think.
Laboratory accidents
With (?)’ investment in research training and workforce development, taking steps to ensure the safety of our trainees and other laboratory workers is of utmost importance.
NIGMS
is providing supplements to training grants for developing curricular materials related to safety. Those resources will be added here as they are developed.
NIGMS
In addition to physical safety in laboratories, (?) is also committed to safety from harassment, be it physical, sexual, or any other inappropriate behavior.
NIGMS
The collection of resources below from within and outside of the United States government may be helpful to (?), as well as funded others in the research community.
NIGMS researchers and trainees
(?) have also authored a perspective in Molecular Biology of the Cell focused on strategies for improving laboratory safety
NIGMS staff
Educational institutions and biotechnology companies use a. (?) of highly hazardous materials.
wide assortment of
When working with these materials every day, it is easy to forget about the harm these materials can cause if they are (?).
stolen
Following the terrorist attacks of September 2001 and the (?) sent the same month, much attention has been directed to practical measures that will keep hazardous materials (biological and chemical) out of the hands of terrorists and criminals.
“anthrax letters”
Many (?) were enacted in response to these terrorist attacks.
new federal laws
It is important that (?) take specific actions to prevent unauthorized entry to labs, secure highly hazardous materials against theft, and ensure compliance with new security regulations.
laboratory personnel
It is essential to implement procedures necessary to provide (?) of all hazardous materials in their areas of responsibility.
security
One objective is to minimize the risk of theft, especially during that five-minute window when the lab is left unattended.
One easy way to increase security is to make sure that your laboratory door is locked whenever the lab is left unattended, even for a few minutes.
Having multiple locked door layers, such as in our laboratory where the chemicals are locked away in a preparation room is very practical in avoiding theft of hazardous material.
Different laboratories implement various security measures which include locking up controlled substances, balances, computers, equipment and syringes and needles.
Laboratory personnel should review and assess the security of their highly hazardous materials, such as infectious agents, toxins, radioactive materials, acutely toxic chemicals, carcinogens, explosive or reactive chemicals, and compressed gases.
The following guidelines were adapted from Appendix F of the CDC/NIH publication, Biosafety in Microbiological and Biomedical Laboratories. The guidelines are intended to reduce the risk for unauthorized removal of hazardous materials from your laboratory:
- Recognize that laboratory security is related to but different from laboratory safety and develop a site-specific security policy. Security, as used in this discussion, refers to measures used to control access to the laboratory in order to prevent theft of materials or equipment from the lab.
o Assess your laboratory for hazardous materials and particular security risks.
o Develop and implement lab security procedures for your lab group.
o Train your lab group on these security procedures and assign responsibilities.
- Control access to areas where hazardous materials are used and stored.
o Close and lock laboratory doors when no one is present. Consider the use of cardkeys or similar devices when the risk warrants.
o Do not leave hazardous materials unattended or unsecured at any time.
o Lock freezers, refrigerators, storage cabinets, and other equipment where biological agents, hazardous chemicals, or radioactive materials are stored when they are not in use.
- Know who is in your laboratory area.
o Consider using a logbook for staff to sign in and out of the lab each day or using carded access devices for this purpose.
o Limit laboratory access to those individuals who need to be in the lab.
o All lab workers (including students, visiting scientists and other short-term workers) should wear identification badges.
o Restrict off-hours access to individuals authorized by the principal investigator.
o Guests should be issued badges and escorted to and from the lab. Approach people you don’t recognize who appear to be wandering in laboratory areas and ask if you can help direct them.
- Know what materials are being brought into your lab.
o Know what hazardous materials are being ordered and shipped to your lab.
o Get rid of unneeded hazardous materials.
o Use a log to sign highly hazardous materials in and out of secure storage.
o Take periodic inventory of all highly hazardous chemicals, biological agents/toxins, radioactive materials, and controlled substances.
- Know what materials are being removed from your lab.
o Track the use and disposal of hazardous materials.
o Require written permission prior to removal of highly hazardous materials from the lab.
o Report any missing inventory.
There is no single simple formula for working safely in the laboratory, since each lab facility and each experiment presents unique challenges. We will be addressing safety issues with each experiment that we do in this course and give you some specific guidelines for safety throughout the semester.
While each chemical that you use will have its own unique properties, there are some common practices that will aid you in treating them all with the level of respect that they are due.
For example, labeling each chemical is required under the law and should be thorough enough so that even a person who does not work in the lab can identify any chemical.
Also, every chemical in the laboratory should have a Material Safety Data Sheet (MSDS) on file and readily available.
The MSDS is a legally required technical document, provided by chemical suppliers, that describes the specific properties of a chemical.
Besides the MSDS on file in the lab, several web sites offer MSDS databases. They are all broken down to the same 8 sections:
The manufacturer‘s contact information is here, along with contacts for emergency situations.
- Chemical identity
Some reagents have multiple components, and many single component chemicals have alternative names. These are all listed here.
- Hazard ingredients/identity
Concentration limits for airborne exposure to a chemical are listed here.
- Hazard ingredients/identity
Although these indices of toxicity are mainly of concern for production workers in factories, they are also useful for evaluation of short-term exposures
- Hazard ingredients/identity
is the maximum airborne concentration of a substance to which workers can be repeatedly exposed without adverse effects.
- Hazard ingredients/identity
TLV (threshold limit value)
The units used are usually
- Hazard ingredients/identity
parts per million (ppm) or mg/m3
This list of physical properties tells you whether the chemical is solid or liquid and how volatile it is.
- Physical chemical characteristics.
This is of particular interest in cases where fire-fighting methods must be selected.
- Fire and explosion hazard data.
This information is essential in determining the proper handling and storage of chemicals.
- Reactivity data.
By knowing the (?) of a chemical, you know what substances or conditions from which you must isolate the chemical.
- Reactivity data.
reactivity patterns
For example, acids and bases react with each other rapidly, giving off large amounts of heat, so should not be stored next to each other.
- Reactivity data.
Others react with water and should be stored in
- Reactivity data.
sealed containers with desiccants.
The best source of specific toxicology data is given here, such as symptoms of acute damage from exposure and some recommended emergency procedures.
- Health hazards.
If a chemical has been tested for carcinogenicity (cancer-causing potential) that information is listed here.
- Health hazards.
In addition, levels at which a chemical has been found to be lethal (called the [?] for lethal dose for 50% of test animals) is listed here.
- Health hazards.
LD50
Since the (?) is dependent on which type of animal it was tested on, as well as how the animal was exposed to the chemical, this information always requires these specifics.
- Health hazards.
LD50
For example, the lethal dose for chemicals is much lower if (?) than it is if ingested.
- Health hazards.
injected
The most common index reported is the (?) for a rat in (?), administered orally (ingestion).
- Health hazards.
LD50; mg of chemical per kg of animal
For volatile chemicals, the toxicity of breathing it is measured as the (?)(lethal concentration in air for half of the test animals), measured in (?); in all cases, the lower the number for the LD50, the more toxic the chemical.
- Health hazards.
LC50; ppm
This describes how to deal with spills.
- Precautions for safe handling and use.
Specific recommendations for personal protective equipment (PPE) are given here.
- Control measures.
Another quick assessment of a chemical‘s health hazards that is usually available on its container is a rating by the
NFPA Ratings (National Fire Protection Association)
A color-coded diamond shape
NFPA Ratings (National Fire Protection Association)
NFPA Ratings (National Fire Protection Association) lists numbers rating a hazard as:
Blue for health hazard
Red for flammability
Yellow for reactivity
for health hazard
Blue
for flammability
Red
for reactivity
Yellow
– oxidizer compound
OX
– acidic compound
ACID
– basic compound
ALK
– corrosive compound
CORR
– use NO WATER
W
– normal material
0
– slightly hazardous
1
– hazardous
2
– extreme danger
3
– deadly
4
– will not burn
0
– flash point > 200oF
1
– flash point > 100oF
2
– flash point < 100oF
3
– flash point < 73oF
4
– stable
0
– unstable if heated
1
– violent chemical change
2
– shock and heat may detonate
3
– deadly
4
There are generally four routes to exposure to hazardous chemicals that you should keep in mind while handling them:
I
S
I
I
: avoid by the use of fume hoods and masks
y
Inhalation
: avoid by the use of lab coats, gloves, and goggles
Skin & eye contact
: avoid eating or drinking in the lab or leaving the lab without removing gloves and washing hands
Ingestion
: dispose of broken glass and needles properly
Injection
Because chemicals pose so many different kinds of hazards, there are no simple rules of thumb for safe handling of them all except for some common sense measures:
Treat [?] as if they were hazardous until you learn otherwise
Label [?] with contents, including concentrations and date that they were transferred
If a hazardous material is contained, label it with a [?]
[?] your experiment BEFORE doing it, making sure that you will not be combining incompatible chemicals
Clean your bench top [?]
Wash hands often and [?] leaving the lab
Take off lab coats and gloves [?] leaving the lab
Always remove gloves [?] touching phones, doorknobs, light switches, etc.
all chemicals
all containers
warning
Think through
before and after use
ALWAYS before
before
before
Ensure proper waste disposal and labeling. Here are some specific tips for handling the different types of hazardous chemicals:
F
C
R
T
Do NOT heat these reagents unnecessarily, and never in the presence of a flame or source of a spark.
Flammables
In general, only open containers in fume hoods.
Flammables
When storing more than 10 gallons of flammable liquids, a special explosion proof storage cabinet is required.
Flammables
: Wear personal protective equipment (PPE) such as lab coats, goggles and gloves, and always add strong acids or bases to water when making solutions.
Corrosives
Neutralize slowly to avoid rapid generation of heat and gases.
Corrosives
Strong acids and bases should never be stored together.
Corrosives
: Wear PPE such as lab coats, goggles and gloves, and know the reactive properties of the chemical.
Reactive chemicals
Always store oxidizing chemicals away from flammable materials.
Reactive chemicals
: Wear PPE such as lab coats, goggles and gloves, and know the toxic properties of the chemical.
Toxic chemicals
When working with a dry powder, wear a mask to avoid breathing the dust.
Toxic chemicals
Be aware of the waste disposal procedures for unused reagents and materials that come in contact with the chemical.
Toxic chemicals
Here are some of the most common hazardous chemicals that you will encounter in the Molecular biology lab:
– formaldehyde
Carcinogens
– ethidium bromide
Mutagens
– acrylamide
Neurotoxins
– formamide
Teratogens
– acetonitrile
Nephrotoxins
– chloroform
Hepatotoxins
– phenol, strong acids & bases
Corrosives
are readily available on the internet.
MSDS
You can reference many chemical manufacturer websites such as
Fischer Scientific and Sigma
Most ACC biology classes, particularly those with laboratory components, use [?] during instruction in addition to images and models.
actual organisms
[?] used at ACC are fundamental in biology instruction and they are utilized to teach specific skills and knowledge. Their condition and usage varies from course to course. Students will be expected to actively participate in these activities
Organisms
Some organisms are observed [?] while others are dead and preserved in various ways.
alive
Student manipulation of organisms ranges from [?] living organisms to [?] preserved ones.
culturing
dissecting
You will be working with live organisms in many biotechnology labs, so it is important to be able to assess any [?] that they may pose and to treat them accordingly.
biological hazards
In general, a live organism is considered a [?] if its release into the environment could have an effect on the health of the environment in general or humans in particular.
biological hazard
This includes known pathogens to humans, plants, or animals, as well as benign organisms containing [?] that could render the recombinant host dangerous.
recombinant DNA
In fact, the [?] itself should be treated as a biohazard, since it is usually inserted into a vector that could transform organisms in the environment if released.
recombinant DNA
Similarly, [?] of human or animal cells should be treated as a biohazard: while they would not survive if released into the environment, most [?] contain recombinant DNA.
tissue cultures
immortalized tissue culture cells
The routes of exposure to infectious agents are the same as those of hazardous chemicals:
inhalation, contact with eyes and skin, ingestion, and injection.
The same general precautions should be taken in handling biological hazards as the guidelines above for handling chemical hazards, especially [?].
toxic ones
Here are some general practices to maximize biological safety:
[?] to the lab at the discretion of the lab director, and adequately train all lab personnel.
Use [?] at all times, and keep all it inside the lab.
Wash hands [?] handling viable materials and animals, after removing gloves and [?] leaving the lab.
Always remove gloves [?] touching phones, doorknobs, light switches, etc.
Avoid touching your [?] with your hands or gloves.
Keep [?] such as coats and book bags out of the lab or in a designated work area.
No [?]; use mechanical pipetting devices.
Minimize [?] and aerosol production.
Disinfect work surfaces to decontaminate [?] a spill and after each work session.
Disinfect or decontaminate glassware [?] washing.
Decontaminate all regulated waste before disposal by an approved method, usually by [?].
Have an [?] in effect.
Use a [?] when available
Limit access personal protective equipment (PPE)
after; before
before
face
personal items
mouth pipetting
splashes
after
before
autoclaving
insect and rodent control program
laminar flow biological safety cabinet
[?] of recorded laboratory-acquired infections are due to inhalation of infectious particles, so special precautions should be taken to avoid producing aerosols when working with pathogens.
Seventy percent
While performing activities that mechanically disturb a [?], the biotechnologist should make the following adjustments
liquid or powder
- mix only in closed containers
Shaking or mixing liquids
- pour liquids slowly
Pouring liquids
- use only cotton plugged pipettes
Pipetting liquids
- point tubes away when opening
Removing a cap from a tube
- sonicate in closed containers
Breaking cells by sonication in the open
- remove slowly
Removing a stopper or cotton plug
- use tubes with screw cap lids
Centrifuging samples
- cool loop first
Probing a culture with a hot loop
Disinfectants such as [?] are used extensively to decontaminate glassware and work areas
bleach and ethanol
it is important to realize that the effectiveness of disinfectants depends on the [?] you are encountering
type of living microorganisms
enveloped viruses
Least resistant
HIV, Herpes simplex, Hepatitis B
Least resistant
bacteria
Slightly resistant
E. coli, S. aureus
Slightly resistant
fungi
Medium resistance
Candida species, Cryptococcus
Medium resistance
non-enveloped viruses
Highly resistant
Polio virus, Mycobacteria, M. tuberculosis
Highly resistant
spore
Most resistant
B. subtilis, Clostridium species
Most resistant
The disposal of hazardous chemicals is subject to state and federal regulations, and is ultimately overseen by the [?].
Environmental Protection Agency
Extremely toxic chemicals are regulated at low levels, and less toxic chemicals can be disposed of through [?] at higher levels.
city sewer systems
Biological hazards should be contained in autoclave bags made of a [?] that are sealed and autoclaved at high temperatures and pressures to completely kill any live organisms.
high melting point plastic
In our laboratory specific hazardous [?] waste disposal will be discussed at the start of every lab.
chemical and biological
Chemical waste disposal containers can be found in the [?].
fume hood
Always keep the fume hood on and the lids to the [?] on the containers.
chemical waste disposal containers
has produced a video explaining safety rules and regulations.
a. The ACC Safety Committee
b. You must sign an [?] to acknowledge that you understand the safety rules and agree to abide by them before you are allowed to use the laboratory in this course.
ACC Safety Contract
has a safety training sheet your instructor will go through with the class together.
c. The Appendix
Pull this out during your safety training.
c. The Appendix
Write down all the information in the blanks and keep it available in your lab notebook while working in the ACC laboratory.
c. The Appendix
Use the [?] to search for chemical company websites, university departments, or other databases containing MSDS information.
Internet
Locate information for the following 3 chemicals:
N
E
S
, an addictive substance found in tobacco.
a. Nicotine
, a stain commonly used for marking DNA.
b. Ethidium bromide
, table salt.
c. Sodium chloride
For each, find the [?] and whether it is a [?]
LD50 (oral, rat, mg/kg)
mutagen or carcinogen
Find a partner to work with, and select a laboratory exercise together from this lab manual that has a list of chemicals and materials that will be used. Using information from MSDS, collect the following information and compile into one table.
chemical name (trade name)
Physical data (appearance, etc.)
NFPA rating
any health hazards/first aid measures
LD50 (mg/kg, oral, rat) or LC50 (ppm)
Toxicity data (carcinogen, mutagen, teratogen, neurotoxin, nephrotoxin, or hepatotoxin)
waste disposal method/spill procedures
any PPE needed
