Welcome to the Pine Lab

(Click on an image to enlarge)

Jerry Pine Jerry Pine is a Professor of Biophysics, and the Pinelab has pioneered in the development of multielectrode arrays for studies of neuron networks in vitro. The present work of the lab is centered on the use of caged neuron arrays for studies of the development and plasticity of neural networks. Contact: jpmail@capsi.caltech.edu


Introduction

A Neuron Cage The neuron cage, made of parylene plastic, is the product of a long term partnership with the microfabrication lab of Yu-Chong Tai in Electrical Engineering. A neuron placed in the center of the cage will attach and grow processes out through the tunnels. The electrode will provide for long term non-destructive two-way communication with the cell, to stimulate it or record from it.


Cells in a Caged Neuron Array Sixteen cage arrays on a silicon chip have been used in preliminary experiments. The electrode leads connect to pads on the chip and then to a printed circuit board for connection to camputer-driven electonics. The Nomarski photo shows a culture of ten caged neurons after ten days in culture, growing many axons and dendrites that form a rich network.

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Data

Recordings The connectivity of a network is determined by stimulating each neuron and recording from all the rest. The picture shows typical data, from stimulation of the neuron in cage number 7 and resulting action potentials at cages 13, 10, 14, and 2.


Culture Evolution The development of connections over time can be seen in Culture Evolution diagrams, shown here for a culture with thirteen neurons. "DIV" means "days in vitro". The red lines indicate short, presumably monosynaptic, delays and the green lines are consistent with polysynaptic connections.


Culture State At a given time, such as for 22 DIV in the example above, all the connections can be examined, with their delay times color coded, according to the bar at right. Each black square is a stimulated neuron, and the surrounding 4 x 4 squares show the responding neurons and their delay times. When there is a disynaptic connection, the times of the two successive responses are shown, and the total delay time, as parts of the square.

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Experiments

There are four present foci:

  1. The spatial and temporal distribution of developing network connectivity of hippocampal cells.
  2. The specificity of connections between identified hippocampal cell types compared with their in vivo behavior.
  3. The activity-dependent plasticity of network connectivity.
  4. The integration of stem-cell derived neurons with networks of hippocampal neurons.
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Publications


  • Erickson, J. C., Tooker, A., Tai, Y-C., and Pine, J. (2008). Caged neuron MEA: A system for long-term investigation of cultured neural network connectivity. J, Neurosci. Meth. 175:1-16. [pdf]
  • Pine, J. and Chow, G., (2008). Moving live dissociated neurons with optical tweezers. IEEE Trans. Biomed. Eng'g. In press.
  • Wagenaar, D. A., Pine, J., and Potter, S. M. (2006). Searching for plasticity in dissociated cortical cultures on multi-electrode arrays. JNRB 5:16-35.
  • Wagenaar, D. A., Pine, J., and Potter, S. M. (2005). An extremely rich repertoire of bursting patterns during the development of cortical cultures. BMC Neuroscience 7:11-29.
  • Wagenaar, D. A., Madhaven, R., Pine, J., and Potter, S. (2005). Controlling bursting in cortical cultures with closed-loop multi-electrode stimulation. J. Neurosci. 25:680-688.
  • Tooker, A., Meng, J., Erickson, J., Tai, Y-C., and Pine, J. (2005). Biocompatible parylene neurocages. IEEE Engineering in Medicine and Biology 24:30-33.
  • Pine, J. (2005). A History of MEA Development. In Advances in Network Electrophysiology Using Multielectrode arrays. M. Taketrani and M. Baudry, Eds., Kluwer, NY.
  • Wagenaar, D. A., Pine, J., and Potter, S. M. (2004). Effective parameters for stimulation of dissociated cultures using multi-electrode arrays, J. Neurosci.. Meth. 138:27-37.
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