1. Fluorescence-based nanosensors measuring oxygen fluctuations during seizure like events Justin Ingram Neuroscience Graduate Student

2. 1 st step: Generate seizure events in hippocampal slices with 4-AP www.anat.ucl.ac.uk/.../okeefe_research.html Ziburkus et al. 2006

3. The 3 month seizure road block -mothers milk (anti-seizure effect) -diet (low ketone diet) -temperature (34°C-35°C) -pH (7.2) Parameters we changed: -4 AP conc. (0uM-200um) -Na +,K +,Ca 2+,Mg 2+ (incubation/recording chamber) -slice thickness (300um-600um) -agarose support blocks (50um change = no seizure) -age (p21)

4. Step 2: Record from hippocampal interneurons and pyramidal cells

5. Creating double transgenic mice which express GFP basket cells (glowing interneuron) Parv/CRE LacZ/GFP Parv/GFP Express GFP on Parvalbumin cells  Glowing Interneurons (Basket Cells) http://www.scq.ubc.ca/wp-content/crelloxmousefeeding.gif

6. Step 3: Use nanometer liposomes to measure oxygen fluctuations in pyramidal/basket cells (GFP labeled) Pyramidal Basket Cell (GFP) OLM -horizontal

7. Fluorescence to measure membrane potential using extracellular dye (Di-8) KCl K+K+ K+K+ K+K+ K+K+ valinomycin K+K+ K+K+ K+K+ K+K+ FL Ratio (UV/Blue-Green) Image plotted with Origin