1. Lab 1: Project Overview and Establishing Group Work Norms

    1. Project overview and schedule
    2. Establishing groups and group work norms

  2. Lab 2: Understanding & Visualizing Perineuronal Nets (PNNs)

    1. Introduction to perineuronal nets
    2. Reading and preparing to present papers
    3. Short informal group presentation on PNNs
    4. Introduction to immunohistochemistry and how it works
    5. Example paper using IHC to visualize PNNs
    6. Write out an IHC/IF protocol for PNNs

  3. Lab 3: Getting to know the mouse brain (Wet Lab)

    1. Using the Allen Brain Atlas
      1. Survey your brain region of interest
      2. Checking out database of existing structures
    2. Update protocol and select antibodies/labels
    3. WET LAB: fresh brain tissue dissection to understand the brain

  4. Lab 4: Developing a hypothesis

    1. Groups do literature review and develop a hypothesis for their project.
    2. Guiding questions:
      1. What brain region would you expect to see differences in PNN expression due to housing condition?
      2. What is the nature of the expected differences? (e.g. more PNNs? fewer PNNs? thinner or thicker PNNs? subregion differences?)

  5. Lab 5: IHC from slice to slide; learning to slice (Wet lab)

    1. Explore the steps from brain harvest to slide mounting (in theory)
    2. Cryoprotection, frozen embedding in OCT, cryosectioning, incubation, and mounting onto slides

  6. Lab 6: Slice your brains!

    1. You will be cryosectioning your brains
    2. you will store your brain slices in 0.1% sodium azide solution

  7. Lab 6.5 (out of class): Blocking and first incubation

    1. Begin IHC with washing away 10% sodium azide solution
    2. Incubate in WFA overnight

  8. Lab 7: Second incubation and mounting onto slides

    1. In class, second incubation
    2. Mount the tissue onto slides and store in dark

  9. Lab 8: Getting to know the confocal microscope

    1. Lecture on confocal microscopy and how it works
    2. Lab Instructor or lab tech will take student groups in turns to learn about the confocal. Meanwhile waiting groups will work on a set of questions to confirm their understanding.
    3. Image a sample brain slice

  10. Lab 9: Imaging your tissue

    1. You and your team on the confocal!
    2. Give yourself about an hour or so. You can use class time that week, usual lab time, or schedule some time outside of class.

  11. Lab 10: Image Analysis & Reporting

    1. Work on your posters