A microfluidic chip for axonal isolation and electrophysiological measurements

A microfluidic chip for culturing neurons and spatially isolating axons from somas is presented for use with visually guided whole-cell electrophysiological measurements. A modular design consisting of detachable and re-sealable layers is used to satisfy the requirements of both long-term neuron culturing as well as electrophysiological measurements. Whole cell patch clamp recordings indicate functional viability of neurons with isolated axons. Fluidic isolation was used to achieve asymmetric lentiviral infection of neurons on a single side reservoir. Neurons were asymmetrically infected with lentiviruses expressing the light-activated cationic channel channelrhodopsin-2. Light-evoked excitatory postsynaptic responses were detected by whole cell recordings of neurons on the uninfected side showing functional synaptic connectivity between the two isolated but axonally connected sides of the device.

[1]  Justin C. Williams,et al.  An automated microdroplet passive pumping platform for high-speed and packeted microfluidic flow applications. , 2010, Lab on a chip.

[2]  S. Franssila,et al.  Complex Droplets on Chemically Modified Silicon Nanograss , 2008 .

[3]  Feng Zhang,et al.  Multimodal fast optical interrogation of neural circuitry , 2007, Nature.

[4]  Digant P. Dave,et al.  Neuro-optical microfluidic platform to study injury and regeneration of single axons. , 2009, Lab on a chip.

[5]  Noo Li Jeon,et al.  Neurotrophin-mediated dendrite-to-nucleus signaling revealed by microfluidic compartmentalization of dendrites , 2011, Proceedings of the National Academy of Sciences.

[6]  W van der Wijngaart,et al.  A High-Yield Process for 3-D Large-Scale Integrated Microfluidic Networks in PDMS , 2010, Journal of Microelectromechanical Systems.

[7]  Jean-Louis Viovy,et al.  Axon diodes for the reconstruction of oriented neuronal networks in microfluidic chambers. , 2011, Lab on a chip.

[8]  G. Whitesides,et al.  Poly(dimethylsiloxane) as a material for fabricating microfluidic devices. , 2002, Accounts of chemical research.

[9]  K. Deisseroth,et al.  Millisecond-timescale, genetically targeted optical control of neural activity , 2005, Nature Neuroscience.

[10]  Nitish Thakor,et al.  Circular compartmentalized microfluidic platform: Study of axon-glia interactions. , 2010, Lab on a chip.

[11]  Nitish Thakor,et al.  Valve-based microfluidic compression platform: single axon injury and regrowth. , 2011, Lab on a chip.

[12]  E. Schuman,et al.  Postsynaptic Decoding of Neural Activity: eEF2 as a Biochemical Sensor Coupling Miniature Synaptic Transmission to Local Protein Synthesis , 2007, Neuron.

[13]  Shelly E. Sakiyama-Elbert,et al.  A microdevice platform for visualizing mitochondrial transport in aligned dopaminergic axons , 2012, Journal of Neuroscience Methods.

[14]  Erin M. Schuman,et al.  Microfluidic Local Perfusion Chambers for the Visualization and Manipulation of Synapses , 2010, Neuron.

[15]  Noo Li Jeon,et al.  Micro-scale and microfluidic devices for neurobiology , 2010, Current Opinion in Neurobiology.

[16]  Noo Li Jeon,et al.  Microfluidic Multicompartment Device for Neuroscience Research. , 2003, Langmuir : the ACS journal of surfaces and colloids.

[17]  C. Rivera,et al.  Expression of GluK1c underlies the developmental switch in presynaptic kainate receptor function , 2012, Scientific Reports.

[18]  A. Scherer,et al.  Applications of microfluidics for neuronal studies , 2007, Journal of the Neurological Sciences.

[19]  E. Bamberg,et al.  Light Activation of Channelrhodopsin-2 in Excitable Cells of Caenorhabditis elegans Triggers Rapid Behavioral Responses , 2005, Current Biology.

[20]  Bruce C Wheeler,et al.  Novel MEA platform with PDMS microtunnels enables the detection of action potential propagation from isolated axons in culture. , 2009, Lab on a chip.

[21]  Noo Li Jeon,et al.  Axonal elongation triggered by stimulus-induced local translation of a polarity complex protein , 2009, Nature Cell Biology.