Transcript Zombie Apocalypse: Identifying Patient Zero - Bio-Rad

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.
[email protected]
 Damon Tighe
[email protected]
 Leigh Brown, M.A.
[email protected]
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TV or real life?
The Walking Dead
show
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Feb 11, 2014 Atlanta
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
C. Macrophage
A. Pathogen
Zombie virus
D. Macrophage
B. B cells
F. T cell
E. Macrophage
G. B cell
J. Antibodies
attach to pathogen
H. Memory B cells
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I. Plasma cells
Antibodies are ineffective
Against Zombie Virus
Crash Course in Immunology and ELISA
Antibody Structure
ELISA tests are
based on immune
system antibody
molecules
Heavy
chain
Disulfide
bonds
Light
chain
Antigens
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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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ELISA Time!
Testing for who is infected
Label Tubes
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Incubate: 5 minutes
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Tap out liquid
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ELISA Time!
Testing for who is infected
Wash with Buffer (contains Tween20)
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Tap out liquid
Repeat
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ELISA Time!
Testing for who is infected
Add 50 µl + Control
Add 50ul – Control
Add 50ul of Bodily Fluid
Your ID #
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Your ID #
ELISA Time!
Testing for who is infected
Incubate: 5 minutes
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Protein (Antigen)
H
R
H
R
Hydrophobic
interactions
Polystyrene (plate)
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ELISA Time!
Testing for who is infected
Tap out liquid
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ELISA Time!
Testing for who is infected
Wash with Buffer (contains Tween20)
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Tap out liquid
Repeat
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ELISA Time!
Testing for who is infected
Add 50 µl “PA” (Primary Antibody)
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Incubate: 5 minutes
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Variable
region
Heavy
chain
Light
chain
constant
region
3,368,600 antibody combos
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ELISA Time!
Testing for who is infected
Concept
Corner
Antigen = chicken gamma globulin
Primary Antibody = anti-chicken
polyclonal antibody
Secondary Antibody =
goat anti-rabbit conjugate to HRP
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ELISA Time!
Testing for who is infected
Tap out liquid
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ELISA Time!
Testing for who is infected
Wash with Buffer
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Tap out liquid
Repeat
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ELISA Time!
Testing for who is infected
Add 50ul “SA” (Secondary Antibody)
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Incubate: 5 minutes
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Amplification of signal
Introduce HRP
(Horseradish Peroxidase)
for colormetric detection
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
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ELISA Time!
Testing for who is infected
Tap out liquid
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ELISA Time!
Testing for who is infected
Wash with Buffer
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Tap out liquid
Repeat x2
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ELISA Time!
Testing for who is infected
Add 50 µl “Sub” (HRP substrate)
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Incubate: 5 minutes
Watch for color change!
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
ELISA Time!
Testing for who is infected
Concept
Corner
Your
ID #
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Your
ID #
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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Deducing who was originally infected
Who is left?
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Guest Zombie Instructor
 Jeff Lukens
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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???
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