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Multielectrode Array

Multielectrode Array. Membrane Biophysics 9 November 2007 John Corthell and Kristal Tucker. Two broad categories of multielectrode recordings. In vivo - KT Recording and stimulation Acute and Chronic Heart, CNS, PNS and Retina In vitro - JC Organotypic and primary dissociated cultures

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Multielectrode Array

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  1. Multielectrode Array Membrane Biophysics 9 November 2007 John Corthell and Kristal Tucker

  2. Two broad categories of multielectrode recordings • In vivo - KT • Recording and stimulation • Acute and Chronic • Heart, CNS, PNS and Retina • In vitro - JC • Organotypic and primary dissociated cultures • Heart, CNS, PNS, and retina

  3. Roadmap • History • Applications • Techniques • Representative articles

  4. Brain-Computer Interface Scott 2006.

  5. Chronic in vivo recordings Musallam et al 2007

  6. Electrode fabrication Musallam et al 2007

  7. Array insertion Musallam et al 2007

  8. Data capture and analysis Musallam et al 2007

  9. Variable depth arrays Sato et. al. 2007

  10. Hochberg et al 2006

  11. Hochberg et al 2006

  12. Hochberg et al 2006

  13. Hochberg et al 2006

  14. Hochberg et al 2006

  15. Hochberg et al 2006

  16. Hochberg et al 2006

  17. In vitro multielectrode array history • Gross, in 1979, first developed an array based on semiconductor technology • Regehr et al., 1989-first applied Aplysia, Hirudo (leech) and Helisoma (snail) cells to multielectrode array (MEA) chip for long-term recording • Masuda et al., in 1983, applied a linear electrode array to myoneural junctions

  18. Linear electrode array recording

  19. Multielectrode array recording

  20. In vitro multielectrode applications • Olfactory processing-Christensen et al., 2000 • Long-term recording-Regehr et al., 1989 • Circadian rhythms-Abraham et al., 2005 • Neuromuscular junction activity-Masuda et al., 1983 • Network analysis • Long-term potentiation • Synaptic interaction

  21. A different type of cell culture that works with MEAs and preserves some circuitry (but not exactly native-synaptic rearrangement) Ideal for long-term recording, as a culture can last from 3-4 weeks for recording to several months, depending on prep Organotypic Slice Culture Duport et al., 1999

  22. Slice cultures preserve 3-dimensional area for electrode preparation Simple to prepare-remove brain (no more than 60s), place into cold solvent, cut into 425m thick slices, place onto MEA with media Spinal cord prep, from Bio-Rad website Organotypic Slice Cultures, cont.

  23. Fabrication • Commercially available, so you don’t have to make one yourself • TiN=titanium nitride

  24. MED is newer than MEA-MED is a planar multielectrode array MED is an attempt to lengthen recording time from previous MEAs Fabrication-MED

  25. Hooked up to amplifier, A/D converter, and computer Typically software programs allow for recording and stimulation near-simultaneously Cells are usually grown in culture dish over the MEA, but can be organotypic Works like most electrophysiology recordings-difference is previous work to set up array and post-experiment work to analyze data MEA/MED Usage

  26. Views of MEA chamber and amplifier plate -PP-probe pin -SC-stimulus connector -RA-recording area

  27. MEA is often used in conjunction with other techniques, such as Ca imaging MEA measures extracellular changes (as you cannot patch), so some things (like post-synaptic potentials and Ca flux) are missed Optical recording techniques (identifying individual cells) are used with MEA to alleviate this MEA + other techniques

  28. Other shortcomings • MEA biochips are expensive to manufacture (may change with time), so researchers will clean the chip to attempt to salvage the product for future use ($250-$350) • Continued cleaning will result in degradation of chip until readings are no longer reliable

  29. Granados-Fuentes et al., “Olfactory bulb neurons express functional, entrainable circadian rhythms.” European J. Neuroscience, 19: 898-906, 2004.

  30. Per1 transgenic rats (yes, rats) underwent bulbectomy from E15-P37, cells were dispersed onto MEAs MEAs had 60 electrodes, spaced 200m apart, with 10m tips (purchased from Germany) SCN explants used as controls (P1-P7) Cultures were covered with a membrane and transferred to a recording incubator Recorded from 4 cultures for at least 5 days simultaneously Used to establish spontaneous activity MEA and setup

  31. Recording apparatus from inside the incubator neuro.gatech.edu/groups/ potter/realtimedac.html

  32. Other techniques used • Locomotor activity measured in normal vs. bulbectomized rats • Per1 activity measured by bioluminescence (Per1 gene is linked to luciferase gene [light from fireflies], add luciferin, and protein product will light up) from a photomultiplier tube • Temperature entrainment via incubator

  33. Per1 expression in OB Start showing rhythm at E19 Results

  34. Top-firing of OB neuron Bottom-firing of SCN control OB neurons that fired rhythmically were found in the mitral cell layer but not the granule cell layer

  35. Left axis is Firing Frequency • Different cells in the same culture can have different firing rhythms

  36. Top-Mitral Bottom-Granule

  37. Removal of OB has no effect on running wheel behavior Temperature changes work as zeitgebers (entraining signals) for OB culture cells

  38. Conclusions from paper • There is a rhythm of activity and Per1 expression in the olfactory bulb neurons of the mitral cell layer • This rhythm begins at E19 and matures over the first week postnatal • These oscillating neurons can have different rhythms from one another in the same culture

  39. Conclusions from in vitro MEA • Most modern MEA is the MED-the planar MEA biochip • Grow cells on biochip or use organotypic culture to study • Can be used to simultaneously record and stimulate extracellularly • Must be cared for-expensive • Should be used with other techniques to compensate for shortcomings

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