Course introduction, Lab Safety

• General overview of the course

• Lab Safety
General overview
Course overview
• BIOL 110L – Modern Biology I
• Core course – 2 credits
• Essential course for consequent years of Undergraduate studies at
NU
• Quizzes, Midterm and Final Exams
• Lab notebooks
Assignments
• Quiz I – 5%
• Quiz II – 7%
• Midterm Exam – 20%
• Quiz III – 8%
• Final Exam – 30%
• Lab Notebook – 20%
• Labster – 10%
• Attendance: class participation is mandatory, missing more than 20%
of Zoom sessions will make you fail the course
Quizzes
• You will have total 3 short quizzes, each include theoretical part of
the previous lab or two
• Question types: multiple correct choice, single correct choice, True-
False and short open answer questions
Exams
• No curving will be applied for this course
• No re-take of the exams unless you have absolutely firm evidence
from the Dean of SSH or other NU authorities
• Final exam cannot be missed, missing this assignment leads to the
failing of the course
Why Labbook?
• Lab notebook is an essential part of any research project
• All labs will have their own labbook filling style and requirements,
but general rules are the same and we will be given them to you
• It is crucial to understand that the labbok is the only evidence of
your work
• Also the documented experiment protocols will help in repeatability
of the project and its key experiments
Starting the Labbook
• During the COVID19 pandemic, you will need to have a digital copy
of the labbook
• These will be the MS Word, Libre Office Writer or similar documents
• For each lab you will need to indicate date, time, and the page
numbers
Labs
• Jan 24-30 - Intro, Lab Safety
• Jan 31-Feb 6 - Lab Math
• Feb 7-13 - Osmosis
• Feb 14-20 - Dialysis
• Feb 21-27 - Cell Microscopy
• Feb 28-Mar 6 - Genes in a Bottle
• Mar 14-20, 28-Apr 3 - Crime Scene Investigator PCR I & II
• Apr 4-10, 11-17 - Forensic DNA Fingerprinting I & II
Academic Integrity
• The NU academic community is absolutely intolerant to cheating and
plagiarism
• “Copied from a friend”, “just copied to formulate my own thoughts”
and other excuses will lead to zero mark for the whole lab
• Please take this message seriously: You are not allowed to copy from
any sources, you are expected to give your own answers
Lab Safety
Why is Lab Safety Important?
• A major aspect of every lab based science class
• It is imperative that everyone starting to work in the lab gets
introduced and trained with lab safety rules
• The rules and symbols are needed so that students do not injure
themselves and their colleagues.
Memorial Wall

Carlos Amaral, 2013 North Andover, MA 51 year old, died from burns after
trimethyliridium explosion
Richard Din, 2012 San Francisco, CA 25 year-old lab worker, exposure to bacteria, died of
septicemia and meningitis
Nilamma, 2011 Mysore, India Died in alcohol fire

Malcolm Casadaben, 2009 Chicago, IL Died from exposure to Yersenia Pestis (plague)
bacterium
Antonina Presnyakova, 2004 Novosibirsk, Russia A lab technician died from Ebola virus infection

In memory of those who have lost their lives in laboratory accidents

General Safety Rules for NU laboratory
• Be prepared: study the laboratory assignment before the lab
• Read the instruction first and follow instructions very carefully
• If you do not know how to do the step/steps of experiment safely,
ask your teaching assistant
• Be able to use all safety devices and protective equipment provided
for your use and know their location (eyewash fountain, safety
shower, fire extinguisher, first aid kit)
• Do not eat, drink or apply cosmetics in the laboratory (and do not
store food in the refrigerators)
• Smoking in the laboratory is absolutely forbidden
General Safety Rules for NU laboratory
• Safety goggles must be worn at all times
• Wear protective clothing all the time during the experiments and
especially when handling corrosive, toxic, or flammable materials
• Avoid loose sleeves, loose cuffs
• Pull back long hair into a ban
• Proper shoes that will cover your feet completely are required (no
sandals, no shoes that are made of fabric, no open toe shoes)
• Ties, scarves, long necklaces, bracelets and dangling earrings should be
removed before starting an experiment
• Piggyback rides or horseplays are prohibited. Do not run in laboratory
areas. Do not push each other. Talk quietly.
General Safety Rules for NU laboratory
• Point away the end of scissors and other sharp objects when walking
• Do not use chemicals from unlabelled containers
• Make sure that once you aliqout, transfer, place a substance in the
new container, you label it
• If you see a colleague doing something dangerous, point it out and
inform the TA
• Report to your TA all unsafe conditions, unsafe acts, that might cause
future accidents
• Report any accident or fire, no matter how trivial
General Safety Rules for NU laboratory
• Wear gloves all time
• Keep your hands in gloves away from your face
• Do not rub your eyes and touch your mouth without washing and
drying your hands first, and of course after taking your gloves off
• Wear glasses rather than contact lenses. However, eyeglasses are
not substitute for proper eye protection
General Safety Rules for NU laboratory
• Be especially mindful of fire hazards when you or your lab neighbors
are working with flammable liquids
• Never leave a reaction or experiment running unattended, unless
you have told your lab partners enough about it to deal with
potential hazards while you are away
• Always close gas valves you opened for your lab work after you
finished their use
• Make sure that Bunsen burners are off before you leave the lab
• Never work alone in the lab at any time of the day even if all lab
safety rules are followed
Routes of Entry for reagents
• Adsorption through the skin
• Wear gloves and lab coats, closed toed shoes
• Splash to the eyes
• Wear safety glasses
• Ingestion into Digestive Tract
• No food or drinks, no chewing on gum or pens
• Injection to the blood stream
• Proper use of sharps
• Inhalation
• Prevent aerosol spraying
• Use Biological Safety Cabinet (also called Hoods)
Goals of Biosafety

Prevent biohazard from harming individuals

• You: Correct PPE and microbiological procedures
• Other lab workers: Promptly lean up spills, and minimize aerosols
• Custodians: Proper waste removal
• Researchers in other labs: No wearing protective equipment outside
of laboratory
What accidents happen in the lab?
• Fire
• Explosion
• Chemical and thermal burns
• Cuts by glass, needles, razors and other sharp objects
• Bites by lab animals
• Absorption of hazardous materials through skin
• Ingestion of hazardous materials by mouth
• Inhalation of toxic fumes
Hazardous materials in the lab
3 main groups:
• CHEMICAL
• PHYSICAL
• BIOLOGICAL
They include materials that are radioactive, flammable, explosive,
corrosive, oxidizing, asphyxiating, biohazardous, toxic, pathogenic, or
allergenic
Chemical hazards
Dangerous chemical materials that are solids, liquids, or gases that can
harm people, other living organisms, property, or the environment
Hazardous chemicals may cause:
• cancer, birth defects, genetic damage,
• miscarriage, injury and death from exposures
Chemicals might be:
• Toxic
• Corrosive
• Reactive
• Flammable
Hazardous Materials Identification
System (Fire Diamond): United
States
GHS Globally Harmonized System of
Classification and Labeling of Chemicals
MSDS – material safety data sheet
• A legally required technical document provided by chemical
suppliers that describes the specific properties of the chemical
• In Kazakhstan, an equivalent of MSDS is a Technical Passport
• MSDS includes:
• physical data (melting point, boiling point, flash point, etc.), toxicity, health
effects, first aid, reactivity, storage, disposal, protective equipment, and spill-
handling procedures
• A file with MSDS information for all reagents that are being used in
the lab must be kept in each laboratory
In any lab you will be using C2H5OH
• Flammable chemical: ethanol
• 70% Ethanol for disinfection
• 100% Ethanol for various experiments such as
nucleic acid cleaning, etc
• Alcohol should never be dispensed or used near
an open flame
Physical hazards
• Thermal such as cold and heat (Liquid Nitrogen, hotplates, Bunsen
Burners, steam from Autoclave)
• Ionizing Radiation (alpha, beta, gamma, X-ray)
• Non-ionizing radiation (electric fields, Infrared, Microwave, magnetic
fields, Radio Frequency, static electric)
• Ultraviolet light
• Vibration (Hand-Arm, Whole-body)
• Noise (including Infrasound, Ultrasound)
• Pressure equipment & gas cylinders
Biohazards
• An agent of biological origin that has the capacity to produce
deleterious effects on humans, i.e. microorganisms, toxins and
allergens derived from those organisms; and allergens and toxins
derived from higher plants and animals
Biohazards
• Infectious agents and pathogens:
• Bacteria, virus, parasites, fungi
• Human-derived tissues, cells, body fluids
• Non-human primate tissues, cells, body fluids
• Animals – wild trapped or lab stock
• Biological toxins:
• Botulinum, tetrodotoxin, ricin, etc.
• Recombinant DNA, RNA:
• Plasmids, linear naked DNA, synthesized oligos, etc.
• Viral vectors:
• Adenovirus, MuLV, lentivirus, etc.
• All are designed to express transgenes. Many insert in the genome. Modifications like VSV
G can increase your risk.
Biosafety Levels
BSL-1
• Defined organisms not known to cause disease in healthy adults
• Examples: Lactobacillus, E.coli, Bacillus subtilis, Baculovirus
BSL-2
• Moderate risk agents present in the community, that cause diseases
of varying severity
• Examples: Salmonella, Clostridium difficile, Hepatitis Viruses, Herpes
simplex
Biosafety Levels
BSL-3
• Indigenous or exotic agents, aerosol transmission. Cause serious and
potentially lethal infection
• Examples: Brucella, Rift Valley Fever virus, VEE
BSL-4
• Dangerous or exotic, high-risk agents that cause life threatening
diseases
• Examples: Ebola, Marburg, Lassa, Machupo viruses
 BSL-1
Standard practices Special Practices Safety equipment Facilities
Handwashing; None PPE Doors for access
decontamination of BS Cabinet not control
work surfaces, required Sink
culture; no eating, No carpets, no rugs
mouth pipetting Windows opening
to the exterior
 BSL-2
Standard practices Special Practices Safety equipment Facilities
Same as in BSL-1 Anyone entering the PPE Lab doors are self-
lab must be notified closing and have
of hazards Procedures involving locks
splashes and aerosol
Potentially infectious must be conducted Sink near the exit
materials must be in BSC, physical door
placed in a durable, containments Lab windows
leak proof container opening to the
exterior are NOT
recommended
 BSL-3
Standard practices Special Practices Safety equipment Facilities
Same as in BSL-1 Same as in BSL-2 Protective clothing Addition to BSL-2
(coveralls, standards:
wraparound *restricted lab access
gowns) *2 self-closing doors
*anteroom for changing
ALL procedures clothes
must be conducted *windows must be
in BSC (Class II or sealed
III)/physical *ducted air ventilation
containments system
 BSL-4
Standard practices Special Practices Safety equipment Facilities
Enforcement of ONLY authorized One-piece positive Walls, ceiling, floors are sealed
strict access control people can enter pressure supplied air
to the lab suit worn by Double HEPA filters for outdoor
Keep logbook to personnel exhaust air
At the entrance of document people
the lab: Supervisor entering/leaving ALL procedures with Lab rooms designed to ensure exit by
name, BSL 4, agent infectious materials sequential passage (shower, change
information Personnel enter must be conducted room)
and leave through in Class III BSC that is
clothing and linked to double- Entry to lab via airlock with airtight
shower rooms door Doors
Autoclave
How to Eliminate Laboratory Accident
• Train and educate
• Always use forethought and concentration
• Eliminate glass, sharps, and points
• Watch out for that “False Sense of Security”
• Always be careful with needles and syringes
• Always wear personal protective equipment
Personal Protective Clothing
• Lab Coat
• Safety goggles
• Gloves
• MUST wear shoes that cover feet: no open toe shoes and no heels
NU Laboratory SAFETY Equipment
• Personal protective equipment (PPE)
• Fire extinguishers, located at the both entrances in the lab
• Showers, located at the both entrances in the lab
• Eye wash stations/face sprays, one at each sink
• Telephone – TA desks to be used for emergency calls
• Lab safety manual
• Chemical Spill Kit
• Fire alarm in lab and closest alarm in hallway
• First-aid kit
Your Work Station
• Keep your work area uncluttered
• Take to the lab station only what is necessary
• Keep hood and bench areas clean and workable space maximized
• Clean up your lab area upon completion of the laboratory period
• Clear up waste, deal with washing up and put things away as you
finish with them
• Make sure everything is safe before you leave things unattended
Decontamination of Work Spaces
• Liquid Disinfectants
• 70% Isopropyl alcohol or ethyl alcohol
• Volatile and flammable
• Acts quickly with no residue
• 10% bleach solution
• Must be prepared daily
• Effective against wide range of agents
• Requires contact time to deactivate agent
• Leaves residue
• Disinfect work surfaces daily and after spill
Biological Safety Practices
Leave the bio in the lab
• Wash hands after removing
gloves and before leaving
• Don’t wear lab coat and gloves
outside of the lab