1. Identify Patient Zero of a Zombie Apocalypse With the Power of an ELISA!

2. Instructors - Bio-Rad Curriculum and Training Specialists
Sherri Andrews, Ph.D.
Damon Tighe
Leigh Brown, M.A.

3. TV or real life?
The Walking Dead show
Feb 11, 2014 Atlanta

4. Zombiefication Method of infection Symptoms Treatment
Bodily fluid contact
Airborne
Foodborne
Other?
Symptoms
Undead
Hungry for flesh
Treatment
Under investigation

5. Introduction to Immunology, Antibodies and ELISA
Fluid swapping and disease spreading!
Indirect ELISA to narrow down who were the initial carriers of the disease

6. 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
Antibodies are ineffective
Against Zombie Virus
H. Memory B cells
I. Plasma cells

7. Crash Course in Immunology and ELISA
Antibody Structure
ELISA tests are
based on immune
system antibody
molecules
Light chain
Heavy chain
Disulfide bonds
Antigens

8. 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.

9. Time to Swap Fluids and disease!

10. 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

11. 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

12. 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

13. Testing for who is infected
ELISA Time!
Testing for who is infected

14. ELISA Time! Testing for who is infected
Your ID #
Label Tubes

15. ELISA Time! Testing for who is infected
Your ID #
Incubate: 5 minutes

16. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

17. ELISA Time! Testing for who is infected
Your ID #
Wash with Buffer (contains Tween20)

18. ELISA Time! Testing for who is infected
Tap out liquid
Repeat
Your ID #

19. ELISA Time! Testing for who is infected
Your ID #
Add 50 µl + Control
Add 50ul – Control
Add 50ul of Bodily Fluid
Your ID #

20. ELISA Time! Testing for who is infected
Your ID #
Incubate: 5 minutes

21. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

22. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

23. ELISA Time! Testing for who is infected
Polystyrene (plate)
Protein (Antigen)
Concept Corner
Hydrophobic
interactions

24. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

25. ELISA Time! Testing for who is infected
Your ID #
Wash with Buffer (contains Tween20)

26. ELISA Time! Testing for who is infected
Tap out liquid
Repeat
Your ID #

27. ELISA Time! Testing for who is infected
Add 50 µl “PA” (Primary Antibody)
Your ID #
Your ID #

28. ELISA Time! Testing for who is infected
Incubate: 5 minutes
Your ID #
Your ID #

29. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

30. ELISA Time! Testing for who is infected
Concept Corner
Your ID #
Your ID #

31. ELISA Time! Testing for who is infected
Variable region
Light chain
Heavy chain
constant region
3,368,600 antibody combos
Concept Corner

32. 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

33. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

34. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

35. ELISA Time! Testing for who is infected
Tap out liquid
Repeat
Your ID #

36. ELISA Time! Testing for who is infected
Add 50ul “SA” (Secondary Antibody)
Your ID #
Your ID #

37. ELISA Time! Testing for who is infected
Your ID #
Incubate: 5 minutes

38. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

39. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

40. ELISA Time! Testing for who is infected
Your ID #
Amplification of signal
Introduce HRP (Horseradish Peroxidase) for colormetric detection
Concept Corner

41. ELISA Time! Testing for who is infected
Concept Corner

42. ELISA Time! Testing for who is infected
Tap out liquid
Your ID #

43. ELISA Time! Testing for who is infected
Wash with Buffer
Your ID #
Your ID #

44. ELISA Time! Testing for who is infected
Tap out liquid
Repeat x2
Your ID #

45. ELISA Time! Testing for who is infected
Add 50 µl “Sub” (HRP substrate)
Your ID #
Your ID #

46. ELISA Time! Testing for who is infected
Incubate: 5 minutes
Watch for color change!
Your ID #
Your ID #

47. ELISA Time! Testing for who is infected
Your ID #
Concept Corner

48. ELISA Time! Testing for who is infected
Concept Corner
Your ID #

49. Deducing who was originally infected

50. Deducing who was originally infected
Everyone Stand UP! If your ELISA was negative sit down

51. Deducing who was originally infected
Round 1
If someone you shared with just sat down, also sit down
Round 2
Round 3

52. Deducing who was originally infected
Who is left?

53. Guest Zombie Instructor
Jeff Lukens

54. Guest Zombie Instructor
Jeff Lukens

55. Using Zombies to Model the Spread of an Infection!
Jeff – a few slides of TI stuff can start here

56. The Zombie Autopsies by Dr. Steven Schlozman, MD

57. 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.

58. 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

59. 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.

60. Normal Dude—Pre-Infection

61. Infected Dude—Early Infection

62. Sick Looking Dude—Stage 2 of Infection

63. Creepy Looking Dude—Stage 3 of Infection

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

65. Graphing the Infection Spread

66. Zooming In For a Better Look

67. Fill the Blocks With Zombies!!

68. Stocking Up For the “Z” Apocalypse!

69. Hmmm….Why are the Graphs Curvy???