biotechnology explorer™ | explorer.bio-rad.com 1 identify patient zero of a zombie apocalypse with...

Biotechnology Explorer™ | explorer.bio-rad.com1

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?

Feb 11, 2014 AtlantaThe Walking Dead

show


Zombiefication

Method of infection– Bodily fluid contact– Airborne– Foodborne– Other?

Symptoms– Undead– Hungry for flesh

Treatment– Under investigation


Introduction to Immunology, Antibodies and ELISA

Fluid swapping and disease spreading!

Indirect ELISA to narrow down who were the initial carriers of the disease


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


Light chain

Heavy chain

Disulfide bonds

Antigens

Antibody StructureELISA tests are based on immune system antibody molecules

Crash Course in Immunology and ELISA


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.


Time to Swap Fluids and disease!


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



Swap with 1 person at the closest table (record who)




Swap with 1 person at the another table (record who)



ELISA Time!

Testing for who is infected


Your ID #

Your ID #

Label Tubes

ELISA Time!Testing for who is infected


Your ID #

Your ID #

Incubate: 5 minutes



Tap out liquid

Your ID #

Your ID #



Your ID #

Your ID #

Wash with Buffer (contains Tween20)




Tap out liquid

Your ID #

Your ID #

Repeat


Your ID #

Add 50 µl + Control

Add 50ul – Control

Add 50ul of Bodily Fluid


Your ID #


Your ID #

Your ID #

Incubate: 5 minutes



Your ID #

Your ID #

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R RHH

Polystyrene (plate)

Protein (Antigen)

Hydrophobic

interactions

Concept Corner



Tap out liquid

Your ID #

Your ID #



Your ID #

Your ID #

Wash with Buffer (contains Tween20)



Tap out liquid

Your ID #

Your ID #


Repeat


Your ID #

Your ID #

Add 50 µl “PA” (Primary Antibody)



Your ID #

Your ID #

Incubate: 5 minutes



Your ID #

Your ID #

Concept Corner



Variable region

Light chain

Heavy chain

constant region

3,368,600 antibody combos

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Concept Corner

Antigen = chicken gamma globulin

Primary Antibody = anti-chicken polyclonal antibody

Secondary Antibody = goat anti-rabbit conjugate to HRP



Tap out liquid

Your ID #

Your ID #



Your ID #

Your ID #

Wash with Buffer



Tap out liquid

Your ID #

Your ID #


Repeat


Your ID #

Your ID #

Add 50ul “SA” (Secondary Antibody)



Your ID #

Your ID #

Incubate: 5 minutes



Your ID #

Your ID #

Concept Corner



Your ID #

Your ID #

Amplification of signal

Introduce HRP (Horseradish Peroxidase) for colormetric detection

Concept Corner



Concept Corner



Tap out liquid

Your ID #

Your ID #



Your ID #

Your ID #

Wash with Buffer



Tap out liquid

Your ID #

Your ID #


Repeat x2


Your ID #

Your ID #

Add 50 µl “Sub” (HRP substrate)



Your ID #

Your ID #

Incubate: 5 minutes

Watch for color change!



Your ID #

Your ID #

Concept Corner



Deducing who was originally infected



Everyone Stand UP!

If your ELISA was negative sit down



Round 1– If someone you shared with just sat down, also sit down




Who is left?



Jeff Lukens

Guest Zombie Instructor


Guest Zombie Instructor

Jeff Lukens


Using Zombies to Model the Spread of an Infection!

Jeff – a few slides of TI stuff can start here


The Zombie Autopsies by Dr. Steven Schlozman, MD


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.


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


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.


Normal Dude—Pre-Infection


Infected Dude—Early Infection


Sick Looking Dude—Stage 2 of Infection


Creepy Looking Dude—Stage 3 of Infection


Trying-to-Eat-You Dude—Full-Blown Infection!!


Graphing the Infection Spread


Zooming In For a Better Look


Fill the Blocks With Zombies!!


Stocking Up For the “Z” Apocalypse!


Hmmm….Why are the Graphs Curvy???

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