1. First Year Experience Research Lab Safety

2. 1. PPE
Personal Protective Equipment: What must be worn when you work in the laboratory.
Eye Protection
Lab Coat
Long Pants
Closed Toed Shoes and Socks – no exposed skin around feet
Lab gloves – when required

3. Eye Protection
Contact lenses are OK as long as glasses/goggles are worn
Prescription glasses – you must wear goggles over them
Safety goggles are provided in organic labs in UV irradiating cabinets
Eye wash stations are present in all labs

4. Clothing and Foot Protection
Clothing must cover all exposed skin including legs/ankles
Socks as required PPE
Stockings or leggings do not
provide good coverage
Sandals, flip-flops, Crocs, open-toe and open-top (i.e. ballet flat) shoes and canvas shoes (i.e. Toms) are not appropriate. These are not going to protect your feet if you drop a piece of glass with a liquid chemical reagent in it.   

5. Be smart about the shoes you elect to wear to lab
This person has on pants and closed toed-shoes but this would not be allowed in lab due to the exposed skin
This person added socks, so this option covers your skin but only offers minimal protection
This option looks better, but imagine chemicals being spilled into the top of these boots
Your best options are sturdy leather footwear that covers the entire top of the foot or a sturdy running shoe.

6. Use of Gloves
Remove gloves before handling objects such as doorknobs, telephones, pens, computer keyboards, pH meter or other electronic buttons, or phones while in lab. It might be convenient to have one gloved hand and one ungloved hand to do procedures where these kinds of things are used.
Throw away gloves anytime you take them off.
You should expect to use several pairs of gloves in any given lab period.

7. Glove Recycling
Contaminated Gloves with visible signs of chemical exposure or those used with
hazardous substances should be collected in solid waste or
Uncontaminated Gloves that have no visible sign of chemical exposure or residue can be placed in
the glove recycling container.
The College is recycling nitrile gloves.  Gloves with visible signs of
chemical exposure or those used with
hazardous substances
should be collected in
solid waste or
biohazard containers.  Uncontaminated gloves with no visible sign of chemical exposure can be placed in the glove recycling container.  
No other waste (paper, etc) can be in the glove recycling boxes.
biohazard containers.
If there’s any questions or the boxes need attention, please contact Dr. Katherine Mullaugh.
Phone:

8. Eyewash / Safety Shower
The safety shower is on the right. Pull the handle and water will start spraying from the shower head on the ceiling. There’s no drain in the floor – we only do this in emergencies, because a flood of water will have to be cleaned up.
The eyewash is on the left. Pull the handle and a fountain of water will appear that you can use to bathe your eyes.

9. Eye Wash

10. Safety Shower

11. Using the Fume Hoods properly
This
window/bar
is called
the sash.
If this is not saying NORMAL, then the hood
is not protecting you. Keeping the sash
and sliding panels in proper position keeps this
NORMAL, otherwise the alarm goes off.
If the alarm goes off, you need to reposition
things to the correct positions, then press the
“mute” button to reset the controller.
The sash should never be raised above
the green “operation” level when you
are working in the hood.

12. ✓ ✓ × ✓ In use, side-to-side panel used as shield Closed, not in use
In use, sash (window) raised
to less than 18 inches
Don’t open side shields
to make one big window. × ✓

13. When using a laboratory hood, Check that the airflow is in the normal range on the digital display
Turn on the hood light
Set the equipment and chemicals back at least 6 inches.
Never lean in and/or put your head in the hood when you are working. This is worse than doing the experiment with no hood at all.
It’s a good idea to put liquid reagent containers in trays to catch all spills and drips

14. Fire Alarms – know the location of one close to your lab

15. Fire Extinguishers – we have several in the labs and in the hallways.

17. Types of Fire Extinguishers
This is a special fire extinguisher for combustible metal fires. It is a type D fire extinguisher. You won’t need to use this unless you work in a research lab with combustible metals.
Most of our fire extinguishers are ABC.
It contains a dry powder to put out the kinds of fires we might encounter in the chemistry labs where we have class.

18. Student Reaction in a Fire
Although we want you to be informed on the operation of a fire extinguisher, we do not expect you to use it. If a fire is ignited in your area, the proper STUDENT response is to:
Notify everyone in the room
If possible shutdown any reaction in progress by removing heat/energy source
Proceed to the nearest exit and pull the nearest fire alarm
Evacuate the building
Assemble in front of the library or in the YWCA parking lot for a positive headcount

19. Keep your lab area clean.× ×
Don’t block the floor in front of the eyewash/shower station.
Don’t leave cords dangling because someone will trip over them. × ×
Don’t leave things in the floor because someone will trip over it.
Throw away used paper towels and used gloves, immediately.

20. Injury procedure
First Aid kits are available in the lab with band aids and other items for treating small cuts and burns.
Campus public safety can be reached at for non-emergencies.
If it is a serious injury, call 911 for emergencies.
The LiveSafe app can also be used to report emergencies and non-emergencies.
Minor cuts or burns can be treated at the discretion of the instructor, but campus safety officials will be called if the instructor thinks it is necessary.

21. Centrifuge Safety

22. Rotor Safety Do not run rotors above their rated speed
Inspect rotor for imperfections and signs of wear that can eventually lead to catastrophic failure
Do not drop rotor
Rinse the rotor after every use
Avoid using abrasive brushes for cleaning
If you suspect rotor has been damaged, do not use it
Do not use a rotor that is not compatible with your model centrifuge
Use tubes and adapters that are rated for use in the rotor being used
Disposable tubes
Fixed angle rotor
Swinging
Bucket
rotor
Need adaptors

23. Accident involving improper rotor usage
Centrifuges that malfunction can create projectiles out of the
rotor shards. If the centrifuge starts to make horrible noises,
cut the power and leave the room

24. Loading the Centrifuge
Be certain that tubes are balanced with a partner
Don’t forget to include caps when weighing the tubes for balance
Secure the rotor on the spindle by tightening all knobs on the lid
Tug gently on the rotor to make sure it is secured to the spindle
Do not overfill bottles (3/4 full max)
Counterbalance your
labeled sample
Both knobs are tightened in some
models to secure rotor to the spindle

25. Unloading Centrifuge
Take precautions if biohazards or other hazardous material is used as aerosols can form during vacuum cycles
Clean the chamber from condensation and any spills
Never try to open the centrifuge door before the rotor is done spinning
Never reach a hand or anything else into the chamber when rotor is spinning
Note: it is sometimes difficult to look at a spinning rotor and determine if it is spinning

26. Centrifuge Safety Overview
Note: A modern centrifuge will have low tolerance for
mismatched tubes and will shut itself off if tubes are not balanced
Also, most modern centrifuges will not allow the door to
unlock while the rotor is still in motion.

27. Autoclave Safety

28. What is an Autoclave?
An autoclave is a specialized piece of equipment designed to deliver heat under pressure to a chamber, with the goal of decontaminating or sterilizing the contents of the chamber.

29. Personal Protective Equipment
Autoclaves utilize steam, heat and pressure and therefore the risk of personal injury through scalding, burns and exploding glassware is great.
Personal protective equipment is absolutely required.
1) Safety Glasses
2) Lab Coat
3) Long pants
4) Closed Shoes
5) Socks
6) Long thermal gloves
7) Face shield recommended

30. What can be autoclaved? Cultures and stocks of infectious material
Culture dishes
Tips, pipettes, gloves, paper towels, aluminum foil
Centrifuge bottles
Glassware -- all caps must be loosened
Media and other aqueous solutions
Contaminated solid items

31. What CANNOT be autoclaved?
Solvents or volatiles
Chlorinated compounds (HCl, bleach)
Corrosives
Radioactive material
Some plastics

32. Cycle Differences Fluids must be autoclaved under a “liquid” setting
Items such as pipette tips, test tubes, and centrifuge bottles are run under “dry” or “gravity” setting
The difference in settings is how the cycle is vented
Liquids must depressurize slowly and dry cycles conclude with a vacuum step to draw off condensation

33. Loading and Unloading the Autoclave
All screw caps must be loosened to prevent pressure changes in the glassware that can cause the container to burst
All items should be placed in an autoclave tray to prevent scald burns in the event of a spill
Return autoclave trays promptly so that other users do not skip using a tray because they can’t find one
Don’t skip using a tray
Do not remove liquid that is still boiling
If possible, allow glassware to cool before removing
Loosen cap by several threads

34. Door Safety Never try to open a door that is under pressure
Never try to speed up the venting process by tampering with the door, by turning on and off the machine, etc. Venting takes time.
Know where the pressure gauges are for the instrument you are using
If possible, vent door slowly

35. Autoclaving Waste
This bag is too full
Contaminated pipette tips and solid waste should be sterilized prior to disposal
Collect waste in a special autoclave-safe biohazards bag
Place bag in secondary container
Vent the bag by opening
Do not overfill bag
After removal place entire bag in a new trash bag so that “biohazard” signs are no longer showing
Sterilized waste can go into the normal trash
Autoclave tape can be used to verify heat delivery but it does not guarantee proper sterilization

36. Container Choice
Pyrex glass, metal, polypropylene (PP) plastic and polycarbonate (PC) plastic are best choices
Polyethylene (PE), polystyrene (PS), and high density polyethylene (HDPE) will often melt and make a mess

37. Autoclaving Tips
Add a 2 cm depth of water to trays with glassware; the water helps eliminate air pockets between the tray and the glass and helps prevent glass from breaking
Do not fill liquid past 75% volume
Separate items to increase steam penetration
Increase cycle time for large volumes of liquid
Temperature must be maintained at 121°C for at least 30 minutes for liquid loads

38. Maintenance Report any irregularities to your supervisor
Do not operate if there is a steam outage
Failed runs should be reported and logged

39. Other Biochemistry Safety Concerns

40. Toxic and Health Hazardous Chemicals
Categories:
Irritants
Sensitizers
Corrosives
Carcinogens
Target Organ Effects
Reproductive Health Toxins
Acute Toxins
Physical Health Hazards
Common routes of exposure in the lab are inhalation and skin absorption, while ingestion is less common.
Carcinogens, reproductive toxins, target organ damage
Acute toxicity, fatality
Source: Basic & Clinical Pharmacology, 12e >Chapter 56. Introduction to Toxicology: Occupational & Environmental Daniel T. Teitelbaum, MD*
Categories list is from Harvard Lab Safety Manual
GHS labels
Irritants, sensitizers, acutely toxic
Corrosives

41. GHS Symbols

42. Biological Hazards
Biological hazards are potential sources of infectious agents that could be harmful to human health.
Bacterial, fungal, parasitic, viral, and prion agents.
Sources may include animals, tissues, cells, blood, and nucleic acid samples, including recombinant DNA.

43. High Voltage Techniques
DNA and protein gel electrophoresis combine high voltage with the use of aqueous solutions
Take precautions to avoid electrocution.
Modern gel boxes have electrodes positioned on the lids to drastically reduce the risk of electrocution.
Always secure the gel box lid before turning on the voltage. Turn off the voltage before removing the lid to a gel box.
Match the red and black electrodes to the corresponding red and black outlets on the power supply.

44. Ultracold (-80°C) Freezer Use
Many biological samples and chemicals need to be preserved at temperatures below room temperature
Always consult the label: 4°C (refrigerator), -20°C (conventional freezer), -80°C (ultracold freezer), -196°C (liquid nitrogen)
Cold storage can slow cell death rate, preserve enzyme activity, inhibit contaminating bacterial growth, and prevent degradation.
Use insulated gloves to handle ultracold materials.
Handle glass dewars with caution – danger of exploding glass if they are knocked over and broken.
Keep the -80° freezer closed!

45. Liquid Nitrogen Safety
Liquid nitrogen (LN2) is commonly used to rapidly freeze proteins and bacteria
LN2 rapidly evaporates and can displace air in enclosed spaces causing suffocation
LN2 can cause death of human tissue from extreme cold
Minor contact can cause “burns”
Evacuated glass dewars can sometimes burst unexpectedly
LN2 can condense liquid oxygen

46. Liquid Nitrogen DON’T’s
DON’T use in confined space
DON’T freeze items in centrifuge tubes with snap caps
DON’T transport LN2 in a closed automobile
DON’T transport LN2 in a passenger elevator
DON’T allow a storage dewar to tip over
DON’T leave cold fingers on a vacuum line in LN2 overnight
DON’T use without PPE!
NO!

47. Liquid Nitrogen Do’s YES! YES!
DO use or dispense LN2 only in well ventilated areas
DO ensure glass dewars are taped or wrapped
DO use approved containers only such as a dewar or threaded cryovials for storage
DO make sure any vessel with LN2 is VENTED
DO secure storage dewars against spilling
DO use appropriate PPE which includes:
Face shield (or minimally goggles)
Long thermal gloves
Apron or lab coat
Closed toed Shoes
Socks
Long pants
NO!
YES!

48. Report any concerns
If you have any safety concerns about the lab you are working in or the people working around you, you can contact:
Your lab instructor
Dr. Marcello Forconi– Head of the departmental safety committee
Dr. Pamela Riggs-Gelasco – Department Chair for Chemistry and Biochemistry
Dr. Jim Deavor, Associate Dean of the School of Science and Mathematics.