Practical Biosafety

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Practical Biosafety

• Or All I need to know about safety I learned
  from my students
• (and one very fastidious TA)

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Joanne Manaster

• Laboratory Teaching Specialist
• MCB 315-Mammalian Histology
• BIOE 202/MCB 493JLM-Cell Culture and Concepts of
  Tissue Engineering Laboratory
• Find this lecture on my BIOE website
• http//www.bioen.uiuc.edu/courses/bioe202/
• Under other courses

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Safety rule 1
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Sure, you are safe.

• Even if you think you are not going to have a
  problem (and you dont need goggles, etc)..
• Safety rule 2 you cant trust the person next
  to you!
• especially the crazy classmate next to you who
  thinks he should pick up the centrifuge and turn
  it upside down to get his sample out!

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Safety Rule 3 Dress for the lab

• Flip flops and Birkenstock sandals are the enemy

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Safety rule 3 ½

• Be prepared when you come to lab
• Know what you are going to be doing
• If you have a lab manual, read it!
• Familiarize yourself with equipment and protocol
• You will work more effectively
• More importantly, and selfishly, it makes my job
  easier

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Safety Rule 4 Chemicals can be hazardous!

• Read MSDS
• Material safety data sheet
• Listen to your instructor
• Remember your basic chemistry safety instructions
• READ LABELS and remain focused
• Mixing of wrong waste products can create a big
  problem
• Or at least create more paperwork

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Safety Rule 5- Sharp things can cause bleeding

• First aid kits are very handy
• Small cuts are 1 injury in my lab classes
• But still rare
• Use sharps containers appropriately
• Keep plastic pipets out of sharps containers
• Dont reach into the sharps containers without
  gloves
• Carefully shake down the container before calling
  for sharps collection
• Broken glassware into a box and into regular
  trash.

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Safety rules 6 Hot items can burn 7- Electric
equipment can shock

• Burns caused by retrieving sample from boiling
  water is the 2 injury in my classes
• Watch orientation of electrodes on
  electrophoretic devices
• Watch for water near electric appliances.

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Safety rule 8- Clean up after yourself

• Reduced clutter
• Improve visibility
• Work efficiently
• Keep yourself and others safe
• Especially important in biology labs
• Biological samples will degrade (and smell)
• Degrading things attract bacteria, mold, etc.
• will ruin future experiments
• a biohazard

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Safety rule 9- No food or drinks in the culture
room!

• Includes drinks, gum, mints, make-up.
• Saliva and crumbs from you can get on your work
  samples
• Your work samples or their by-products can get on
  your food!
• http//www.hersheys.com/reeses/home.asp

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Invisible life forms are everywhere!

• Present on skin, fall off of hair, clothing, in
  saliva, tears, breath.
• Present on surfaces and in water
• Especially warm water

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Safety rule 10- Protecting your cultures
protects you!

• Biosafety hood
• Aseptic technique
• Prevents introduction of organisms into your
  culture
• handwashing
• 70 ethanol
• Sleeves rolled up, hair pulled back, no watches,
  rings, etc., gloves by suggestion

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Safety rule 11- Proper biological disposal is
absolutely essential!

• For cell cultures (Biosafety level 1)
• 10 sodium hypochlorite
• Bleach
• Wait a few minutes and then rinse down sink
• Bacterial and fungal cultures
• Autoclave
• Use autoclave bags

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Biosafety level 2 cautions

• Any human fluid or cells fall into this category
• You are safer working with someone elses cells
  than your own
• Immune system recognizes non-self faster
• Wear gloves, lab coat, goggles/mask if desired
• Separate trash
• Autoclave all disposable pipets in autoclave bag
• Put in dark trash bag and place in regular trash
• Sharps still go in sharps box

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Biological reagents and toxins

• Mitomycin C
• Mitotic inhibitor
• Insulin
• Cholera toxin
• Serum
• Used as a supplement for cell cultures
• Not screened for every disease

• Treat like biosafety level 2
• Wear gloves!
• Separate disposal
• Vials, pipets, etc into autoclave bag
• Not regular trash
• Autoclave then place in dark bag and throw in
  trash

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Compressed gasses

• Make certain that gas tanks are chained to wall.
• Moving gas tanks can be tricky and even dangerous
• Wear goggles when using compressed gas

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Cryo-worksuper cold things can injure you, too!

• Use gloves, when handling dry ice or retrieving
  samples from a -80oC freezer or liquid nitrogen
• wear closed toe shoes when getting liquid
  nitrogen
• Wear goggles when thawing samples frozen in
  liquid nitrogen
• Pressure increases and may blow top off the tube

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Other safety measures

• TAs are variable in what they remember to tell
  you.
• Check for fire extinguisher, shower, first aid
  kit yourself