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Fluorescence-based nanosensors measuring oxygen fluctuations during seizure like events

Fluorescence-based nanosensors measuring oxygen fluctuations during seizure like events. Justin Ingram Neuroscience Graduate Student. 1 st step: Generate seizure events in hippocampal slices with 4-AP. Ziburkus et al. 2006. www.anat.ucl.ac.uk/.../okeefe_research.html.

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Fluorescence-based nanosensors measuring oxygen fluctuations during seizure like events

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

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

  3. Parameters we changed: -4 AP conc. (0uM-200um) -Na+,K+,Ca2+,Mg2+ (incubation/recording chamber) -slice thickness (300um-600um) -agarose support blocks (50um change = no seizure) -age (p21) The 3 month seizure road block -mothers milk (anti-seizure effect) -diet (low ketone diet) -temperature (34°C-35°C) -pH (7.2)

  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) K+ K+ KCl valinomycin K+ K+ K+ K+ K+ K+ FL Ratio (UV/Blue-Green) Image plotted with Origin

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