identify patient zero of a zombie apocalypse with the power of an elisa!
DESCRIPTION
Identify Patient Zero of a Zombie Apocalypse With the Power of an ELISA!. Instructors - Bio-Rad Curriculum and Training Specialists. Sherri Andrews, Ph.D. [email protected] Damon Tighe [email protected] Leigh Brown, M.A. [email protected]. TV or real life?. - PowerPoint PPT PresentationTRANSCRIPT
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Identify Patient Zero of a Zombie Apocalypse With the Power of an ELISA!
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Instructors - Bio-Rad Curriculum and Training Specialists
Sherri Andrews, Ph.D.
Damon Tighe
Leigh Brown, M.A.
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TV or real life?
Feb 11, 2014 AtlantaThe Walking Dead
show
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Zombiefication
Method of infection– Bodily fluid contact– Airborne– Foodborne– Other?
Symptoms– Undead– Hungry for flesh
Treatment– Under investigation
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Introduction to Immunology, Antibodies and ELISA
Fluid swapping and disease spreading!
Indirect ELISA to narrow down who were the initial carriers of the disease
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Crash Course in Immunology and ELISA
Immune Response
A. Pathogen
C. Macrophage
D. Macrophage
E. MacrophageF. T cell
B. B cells
G. B cell
H. Memory B cellsI. Plasma cells
J. Antibodies attach to pathogen
Zombie virus
Antibodies are ineffectiveAgainst Zombie Virus
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Light chain
Heavy chain
Disulfide bonds
Antigens
Antibody StructureELISA tests are based on immune system antibody molecules
Crash Course in Immunology and ELISA
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ELISA Enzyme-Linked Immunosorbant Assay (indirect)
Add the purified antigen to all the wells. Incubate for 5 min. Rinse
Add serum antibodies (student samples) to the appropriate wells. Incubate for 5 min. Rinse
Add the enzyme-linked antibody to all wells. Incubate for 5 min. Rinse
Add enzyme substrate to all wells. Incubate for 5 min.
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Time to Swap Fluids and disease!
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Swapping Fluids and Disease
Swap with 1 person at your table (record who)
Add your 750 µl together, mix and each take 750 µl back
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Swapping Fluids and Disease
Swap with 1 person at the closest table (record who)
Add your 750 µl together, mix and each take 750 µl back
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Swapping Fluids and Disease
Swap with 1 person at the another table (record who)
Add your 750 µl together, mix and each take 750 µl back
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ELISA Time!
Testing for who is infected
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Your ID #
Your ID #
Label Tubes
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Incubate: 5 minutes
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Wash with Buffer (contains Tween20)
ELISA Time!Testing for who is infected
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ELISA Time!Testing for who is infected
Tap out liquid
Your ID #
Your ID #
Repeat
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Your ID #
Add 50 µl + Control
Add 50ul – Control
Add 50ul of Bodily Fluid
ELISA Time!Testing for who is infected
Your ID #
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Your ID #
Your ID #
Incubate: 5 minutes
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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R RHH
Polystyrene (plate)
Protein (Antigen)
Hydrophobic
interactions
Concept Corner
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Wash with Buffer (contains Tween20)
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
Repeat
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Your ID #
Your ID #
Add 50 µl “PA” (Primary Antibody)
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Incubate: 5 minutes
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Variable region
Light chain
Heavy chain
constant region
3,368,600 antibody combos
Concept Corner
ELISA Time!Testing for who is infected
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Concept Corner
Antigen = chicken gamma globulin
Primary Antibody = anti-chicken polyclonal antibody
Secondary Antibody = goat anti-rabbit conjugate to HRP
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Wash with Buffer
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
Repeat
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Your ID #
Your ID #
Add 50ul “SA” (Secondary Antibody)
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Incubate: 5 minutes
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Amplification of signal
Introduce HRP (Horseradish Peroxidase) for colormetric detection
Concept Corner
ELISA Time!Testing for who is infected
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Concept Corner
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Wash with Buffer
ELISA Time!Testing for who is infected
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Tap out liquid
Your ID #
Your ID #
ELISA Time!Testing for who is infected
Repeat x2
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Your ID #
Your ID #
Add 50 µl “Sub” (HRP substrate)
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Incubate: 5 minutes
Watch for color change!
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Your ID #
Your ID #
Concept Corner
ELISA Time!Testing for who is infected
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Deducing who was originally infected
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Deducing who was originally infected
Everyone Stand UP!
If your ELISA was negative sit down
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Deducing who was originally infected
Round 1– If someone you shared with just sat down, also sit down
Round 2– If someone you shared with just sat down, also sit down
Round 3– If someone you shared with just sat down, also sit down
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Who is left?
Deducing who was originally infected
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Jeff Lukens
Guest Zombie Instructor
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Guest Zombie Instructor
Jeff Lukens
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Using Zombies to Model the Spread of an Infection!
Jeff – a few slides of TI stuff can start here
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The Zombie Autopsies by Dr. Steven Schlozman, MD
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The Scenario A new virus has been discovered that affects
the brains of humans, turning them into zombies! Epidemiologists are trying to figure out what to do to help stop the outbreak. The virus is airborne, being transported in saliva droplets of affected individuals. The zombies are attacking healthy humans as their primary food source and new zombies are showing up at an alarming rate.
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Connections to Solid Science Practices
Asking questions and defining problems– What happens to the brain to cause “zombification”?– How does a disease spread through a population?
Using mathematical and computational thinking– Graphically and numerically evaluate the spread of a disease
in a population
Developing and using models– Brain models– Graphical representations of data
Analyzing and interpreting data– Using mathematics and graphical literacy to evaluate dat
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Symptoms of the Infection
Affected Cerebellum: Zombies clumsily shuffle forward.
Affected Hypothalamus: Zombies have insatiable appetites.
Affected Frontal Lobe: Zombies are poor problem solvers.
Affected Amygdala: Zombies aren't nice. They are full of rage.
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Normal Dude—Pre-Infection
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Infected Dude—Early Infection
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Sick Looking Dude—Stage 2 of Infection
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Creepy Looking Dude—Stage 3 of Infection
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Trying-to-Eat-You Dude—Full-Blown Infection!!
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Graphing the Infection Spread
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Zooming In For a Better Look
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Fill the Blocks With Zombies!!
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Stocking Up For the “Z” Apocalypse!
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Hmmm….Why are the Graphs Curvy???