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Review: Detection
Favored method of detection is Microelectrode Arrays (MEAs). Electrodes fabricated on surface of device Monitor signal externally; doesnt damage cells Neural signals typically in the range of 100s of uVpp Many working neuron-based sensors utilize MEAs Much research focused on improving control of neural growth on MEAs.
Photo Source: Nam, et al., Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures. IEEE Transactions on Biomedical Engineering, 51:1 (2004) 158-165.
Source: Chiappalone et a. Networks of neurons coupled to microelectrode arrays: a neuronal sensory system for pharmacological applications Biosensor and Bioelectronics, 18:5-6 (2003), 627-634
Source: Prasad et al. Neurons as sensors: individual and cascaded chemical sensing Biosensor and Bioelectronics, 19:12 (2004), 1599-1610
Challenges
Controlling interaction of living neuron to device.
Ideal of 1:1 association of neurons to electrodes is difficult to achieve Affects signal-to-noise ratio Affects reproducibility and repeatability of response
Long term maintenance of cells in vitro Stability of device (corrosion, biofouling, etc)
Physical Immobilization
Chemical Patterning
Many Types of SAMs Recent research has focused on using thiols on gold substrates
Early work
Potember (1995)
SAM: n-octa-decyltrichlorosilane Selective UV irradiation to remove/pattern SAM Surface made bioactive using synthetic peptide, covalently attached using a cross-linker 5um line widths Optical microscopy to evaluate neural attachment and growth Cells remained restricted to pattern for over 15 days in culture
Source: Potember et al. Conducting networks from cultured cells on selfassembled monolayers Synthetic Metals, 71(1995) 1997-1999.
Thiol-based SAMs
Structure:
Alkane chain, typically with 10-20 methylene units Head group with a strong preferential adsorption to the substrate used. Eg: Thiol (-SH) head groups and Au(111) substrates Tail group gives the SAM its functionality
Thiols on Au(111)
Thiol head group bonds to the threefold hollow site on gold surface. Van der Waals forces between alkane chain causes them to lie at 30 degree angle
Source: Nam, et al., Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures. IEEE Transactions on Biomedical Engineering, 51:1 (2004) 158-165.
Source: Nam, et al., Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures. IEEE Transactions on Biomedical Engineering, 51:1 (2004) 158-165.
Results
Neurons complied to patters for 2-3 weeks Spontaneous neural activity recorded:
400ms
Source: Nam et al. Epoxy-silane linking of biomolecules is simple and effective for patterning neuronal cultures. Biosensors and Bioelectronics 22 (2006) 589597
Note SAM increased impedance by factor of 2-3 Mean SNR of 6.5 at 2 weeks mean amplitude of extracellular spikes was 25uVpp at 7 DIV and 50uVpp at 20-24 DIV. Background noise 2.9uVpp
Source: Palyvoda, et. al., Culturing neuron cells on electrode with self-assembly monolayer Biosensor and Bioelectronics, 22 (2007) 2346-2350.
350x350um: 57 +/-10 200x200um: 16 +/- 5 100x100um: 4.14 +/- 2 50x50um: 1 +/- 0.87
50x50um pad size comes close to single neuron immobilization, with error. No measurement of electrical activity was performed
Conclusions
Neuron-based biosensors appear poised to become an effective biosensor technology. Challenges:
Stability of culture and micro device Reproducibility of results Maintaining health of cells over long term
Conclusions
SAMs are attractive as a possible solution to these challenges
Relatively simple compared to other options Highly biocompatible Customizable Increases stability of neuron-device interface
But
10-15 years or more of characterization studies of SAMs No fully functional SAM-based biosensor for specific application to date
Questions?
References
Chiappalone, M., Vato, A., Tedesco, M.B., Marcoli, M., Davide, F., Martinoia, S., 2003. Networks of neurons coupled to microelectrode arrays: a neuronal sensory system for pharmacological applications Biosensor and Bioelectronics 18(5-6), 627-634. Chun, C., Hickman, J.J., Wang, W., Gregory, C. Narayanan, S. Poeta, M. "Neuronal cell patterning on covalently bound protein patterns by microcontact printing techniques and the functioning of proteins bound on silane monolayers." Conference Paper. Craighead, H.G., Turner, S.W., Davis, R.C., James, C., Perez, A.M., John, P.M., Isaacson, M.S., Kam, L. Shain, W., Turner, J.N, Banker, G., 1998. Chemical and Topographical Surface Modifcation for Control of Central Nervous System Cell Adhesion. Biomedical Microdevices 1, 49-64. Faucheux, N., Schweiss, R., Lutzow, K., Werner, C., Groth, T., 2004. "Self-assembled monolayers with different terminating groups as model substrates for cell adhesion studies." Biomaterials 25 2721-2730. Griscom, L., Degenaar, P. LePioufle, B., Tamiya, E., Fujita, H., 2002. Techniques for patterning and guidance of primary culture neurons on microelectrode arrays. Sensors and Actuators B 83, 15-21. James, C.D., Spence, A.J.H., Dowell-Mesfin, N.M., Hussain, R.J., Smith, K.L., Craighead, H.G., Isaacson, M.S., Shain, W., Turner, J.N., 2004. Extracellular recordings from patterned neuronal networks using planar microelectrode arrays. IEEE Transactions on Biomedical Engineering 51(9), 1640-1648. Lin, S., Chen, J.J., Liaao, J., Tzeng, S., 2008. "Characterization of surface modification on microelectrode arrays for in vitro cell culture." Biomedical Microdevices 10, 99-111. Maher, M.P., Pine, J., Wright, J., Tai, Y., 1999. "The Neurochip: a new multielectrode device for stimulating and recording from cultured neurons." Journal of Neuroscience Methods 87, 45-56. Naka, Y., Eda, A., Takei, H. Shimizu, N. 2002. "Neurite outgrowths of neurons on patterned self-assembled monolayers." Journal of Bioscience and Bioengineering 94(5), 434-439. Nam, Y., Chang, J.C., Wheeler, B.C., Brewer, G.J., 2004. Gold-coated microelectrode array with thiol linked self-assembled monolayers for engineering neuronal cultures. IEEE Transactions on Biomedical Engineering 51(1), 158-165.
References
Nam, Y., Branch, D.W., Wheeler, B.C., 2006. Epoxy-silane linking of biomolecules is simple and effective for patterning neuronal cultures. Biosensors and Bioelectronics 22, 589597. Palyvoda, O., Chen, C., Auner, G.W., 2007. Culturing neuron cells on electrode with self-assembly monolayer Biosensor and Bioelectronics 22, 2346-2350. Potember, R.S., Matsuzawa, M., Liesi, P., 1995. Conducting networks from cultured cells on self-assembled monolayers Synthetic Metals 71, 19971999. Prasad, S., Yang, M., Zhang, X., Ozkan, C.S., Ozkan, 2003, Electric field assisted patterning of neuronal networks for the study of brain functions. Biomedical Microdevices 5(2), 125-137. Prasad, S., Zhang, X., Yang, M., Ozkan, C.S., Ozkan, M., 2004. Neurons as sensors: individual and cascaded chemical sensing. Biosensor and Bioelectronics 19(12), 1599-1610 Romanova, E.V., Oxley, S.P., Rubakhin, S.S., Bohn., P.W., Sweedler, J.V., "Self-assembled monolayers of alkanethiols on gold modulate electrophysiological parameters and cellular mophology of cultured neurons." Biomaterials 27, 1665-1669 Slaughter, G.E., Bieberich, E., Wnek, G.E., Wynne, K.J., Guiseppi-Elie, A., 2004 "Improving Nueron-to-electrode surface attachment via alkanthiol self-assembly: an alternating current impedance study." Laungmuir 20, 7189-7200. Tooker, A., Meng, E., Erickson, J., Tai, Y., Pine, J., 2004. Development of Biocompatible Parylene Neurocages. Proceedings, IEEE EMBS, San Francisco, September 2004, 2542-2545 Widge, A.S., Jeffries-El, M. Cui, X., Lagenaur, C.F., Matsuoka, Y., 2007. "Aelf-assembled monolayers of polythiophene conductive polymers improve biocompatibility and electrical impedance of neural electrodes." Biosensors and Bioelectronics 22, 1723-1732. Wikipedia Self-Assembled Monolayer. http://en.wikipedia.org/wiki/Self-assembled_monolayer. Accessed 5/15/0008. Website: "Self Assembled Monolayers." Molecular Films and Surface Analysis group at the Laboratory of Applied Physics, Department of Physics, Linkping University in Sweden. http://www.ifm.liu.se/applphys/ftir/sams.html. Accessed 5/16/2008.