Biosensory Laboratory Dr. Vitaly J. VodyanoyDirector Current Laboratory Personnel Mission Membrane Biosensor Molecular Probes Olfaction Links Optical Microscope of High Resolution Patents Selected Publications Current Laboratory Personnel Vitaly J. Vodyanoy, Ph.D., Director Iryna B. Sorokulova, Ph.D., Visiting Professor Ludmila P. Globa, Research Associte IV Feng Xu, Research Associate I April Krumnow, Research Assistant III Oleg Pustovyy, Research Assistant III Patrick Kanju, Postdoc Fellow Randy O. Boddie, Research Assistant II (50%) Nilmini Viswaprakash [back to top] Mission The Biosensor Laboratory directed by Dr. Vitaly Vodyanoy is involved with all aspects of advanced sensors, based upon structural and functional principals of biological membranes. It serves as a focal point for teaming with others in fundamental understanding and development of diverse advanced sensors in areas of environmental monitoring, biotechnology; signature sensing, trace detection, diagnostics, and pathogen sensing. Capabilities include thin film technology with films of organized architecture, biological membrane technology, microelectrode and patch-clamp techniques, fluorescence spectroscopy, high resolution dark-field and atomic force microscopy. The programs and activities in Biosensor laboratory cover three major areas: Membrane biosensor, Molecular probes, and Optical microscope of high resolution. [back to top] Membrane Biosensor Artificial membranes, such as monolayers, multilayer assemblies, and large solvent-free lipid bimolecular membranes, will be used as the medium into which biological receptor macromolecules will be functionally incorporated and/or functionally reconstituted. The proposed studies encompass a range of chemosensor prototypes starting from lipid bimolecular membranes in a simple circuit which measures membrane conductance to a multilayer assembly deposited upon chemically sensitive electronic devices, which in principle allows complex discrimination between different chemical substances to be possible. These studies are designed to provide the necessary basis which will lead to the ultimate development of practical chemosensitive electronic devices which provide both ultratrace sensitivity and selectivity for a defined range of organic molecules. Current projects include detection of molecules in gas by Olfactory Sensors. This work is directed toward the design and construction of a sensor for the detection and identification of specific molecules, and based on their relative size, chemical specificity, and characteristic electrical charge. In this sensor, the relative sizes are distinguished by differences in their diffusion through specially constructed small channels in monolayer films. After the molecules traverse the membrane pores, they impinge on a charge coupled device where they generate a signal proportional to their charge. Unique "signatures," or patterns, of particular molecules and their mixtures will be stored, analyzed, and so the system will "learn" to selectively recognize these molecules. (S. Pathirana, T. Roppel, W.C. Neely, and V. Vodyanoy). Another current project includes a sensor for detection of pathogenic bacteria by using mass sensors coated with specific membrane immobilizing bacterial antibodies or DNAs. Changes in the mass, mechanical modulus, or electrical properties of a surface layer due to binding of specific proteins (or DNAs) produces corresponding changes in the measured parameters of the sensor. Two methods are utilized to immobilize antibodies or DNAs on the sensor surfaces: (1) deposition of a composite lipid-protein monolayer onto a sensor surfaces and (2) using Langmuir-Blodgett technique and biotin/streptavidin couple. (S. Pathirana, B. Chin, J. Barbaree, R. Zee, W. Gale, P. Hsieh, D. Conner, W.C. Neely, and V. Vodyanoy). Basic research associated with the Membrane Biosensor program is focused on the initial chemoreceptive events in the animal olfactory system and to find out how is odorant information translated into electrical events in the olfactory receptor. The work involves the transfer of macromolecules from olfactory receptor cells into artificial bimolecular membranes, monolayer and multilayers, and studying the electrical and chemical properties of the reconstituted systems. Another focal point of this research is the use of functional reconstitution of the olfactory receptor cell to produce an artificial system responsive to small (subnanomolar) concentrations of odorant. The objective of this study is to develop a working prototype of an electrochemical sensor which shares basic molecular mechanisms associated with the initial steps in olfaction. We are also studying glutamate receptors reconstituted in bilayers using a combination of patch clamp and biochemical techniques. The main focus of this work is to determine molecular mechanisms of the specific facilitation of electrical activity of some of these receptors involved in the information storage (memory) in the brain. (S. Pathirana, S. Sinnarajah, D. Srikumar, V. B. Bahr, V. Suppiramaniam, E. Morrison, and V. Vodyanoy).   Figure 1. A KSV 2200LB Langmuir-Blodgett System (KSV-Chemical, Finland) is used for deposition of monolayer with controlled architecture. Multilayers are obtained by successive dipping of the sensors through the monomolecular film deposited at a water-air interface on the Langmuir-Blodgett trough. [back to top] Molecular Probes This program involves a design of solid state optical sensors by incorporating specially designed fluorescent sensor molecules into architecturally organized Langmuir-Blodgett (LB) multilayer systems. LB technique will be used to control the orientation of the probe molecules in a optically favorable manner. These LB systems can be interfaced with opto-electronic devices using optical fibers. With the use of properly designed probe molecules effective optical sensors for inorganic ions and organic molecules can be produced. Current projects includes a class of molecules which is designed and synthesized so that they undergo photophysical changes (fluorescence intensity, excitation and emission maxima, fluorescence lifetimes etc.) upon complexation with a specific cation. In present work we use molecules of N-(9-anthrylmethyl)-N-(octadecylethoxy)ethanolamine(C18AE) (de Silva, A. P.; de Silva S. A. J. Chem. Soc., Chem. Commun. 1986, 1709). In the uncomplexed form, the fluorophore emission of this molecule is quenched due to photo electron transfer from the cation receptor to the fluorophore. But upon complexation with the proper cation this electron transfer process will be blocked and therefore the fluorophore emission can be observed and the intensity of emission will be dependent on the cation concentration. This molecule was synthesized and purified in Dr. S. de Silva's laboratory (Department of chemistry and bio chemistry of the Montclair State University) and its fluorescence intensity is sensitive to the pH of the medium. Using functional interface of optically sensitive molecules with opto-electronic devices highly sensitive and selective sensors with very short response times can be designed for the detection of inorganic ions and organic molecules. (S. Pathirana, L. Hemmer, M. Hartel, W.C. Neely, S. de Silva, and V. Vodyanoy). [back to top] Olfaction Links Olfaction Links I Olfaction Links II [back to top] Optical Microscope of High Resolution An optical system which has the capability of showing biological cells and intracellular structures in the real time in their natural environment with no freezing, dehydration or staining is designed. The microscope being adopted for the reflected light can be used for opaque objects to obtain high resolution images of surfaces. (S. Pathirana, W.C. Neely, and V. Vodyanoy). Figure 3. High resolution dark field microscope equipped with a Naessens dark-field condenser (COSE Corp., Canada).   Figure 4. Photographs of red and white blood cells - panels 1 (Dark field) and 2 (Bright field). Mean diameter of red blood cells is about 7 microns. Objective 100X/1.4, oil, Mercury are light. The sample has been photographed in two minutes after the blood has been drawn and no dehydration, freezing, or staining was applied [back to top] Selected Patents Apparatus and method for the measurement of the aerodynamics of olfaction in animals and man. Detection of food-borne Salmonella contamination by polyclonal antibody assays High-resolution optical microscope In vitro assay to determine the croso-species tissue-binding properties of peptide ligands Ligand sensor devices and uses thereof Method and apparatus for generating a voltage across a membrane Method for the protection of biological materials Methods of forming monolayers and their uses thereof II Use of acacia gum to isolate and preserve biological material [back to top] Selected Publications "Novel metal clusters isolated form blood are lethal to cancer cells."  Alexander M. Samoylov, Tatiana I. Samoylova, Oleg M. Pustovyy, Alexei A. Samoylov, Maria A. Toivio-Kinnucan, Nancy E. Morrison, Ludmila P. Globa, William F. Gale and Vitaly Vodyanoy.  Cells, Tissues Organs 2005; 179(3):115-124, 2005. "Anti-tumor extracts isolated from shark tissue."  A.M. Samoylov, T.I. Samoylova, O.M. Pustovyy, A.A. Samoylov, M.A. Toivio-Kinnucan, N.E. Morrison, L.P. Globa, W.F. Gale, and V. Vodyanoy.  Molecular Biology of the Cell 15:253A-253A 1398 Suppl. S., 2004. "Thermodynamic characteristics of mixed monolayers of Amphotericin B and cholesterol."  Jennifer Cannon Sykora, William C. Neely and Vitaly Vodyanoy.  Journal of Colloid and Interface Science 276(1):60-67, 2004. "Structure and function of long-lived olfactory organotypic cultures from postnatal mice."  E.M. Josephson, S. Yilma, V. Vodyanoy and E.E. Morrison.  Journal of Neuroscience Research 75(5):642-653, 2004. "Solvent effects on Amphotericin B monolayers."  Jennifer Cannon Sykora, William C. Neely and Vitaly Vodyanoy.  Journal of Colloid and Interface Science 269(2):499-502, 2004. “Phage display for detection of biological threat agents.”  Petrenko V.A. and V.J. Vodyanoy.  The Journal of Microbiological Methods 1768; 53:253-262, 2003. “Specific and selective biosensor for Salmonella and their detection in the environment.”  E.V. Olsen, S.T. Pathirana, A.M. Samoylov, J.M. Barbaree, B.A. Chin, W.C. Neely, and V. Vodyanoy.  The Journal of Microbiological Methods 1770; 53(2):273-285, 2003. “Amphotericin B and Cholesterol in Monolayers and Bilayers.”  Jennifer Sykora, Solomon Yilma, William C. Neely and Vitaly Vodyanoy. Longmuir 19:858-864, 2003. "Peptide biosensor for recognition of cross-species cell surface markers.”  Alexandre M. Samoylov, Tatiana I. Samoylova, Suram T. Pathirana, Ludmila P. Globa and Vitaly J. Vodyanoy.  Journal of Molecular Recognition 15:197-203, 2002. “Recognition of cell-specific binding of phage display derived peptides.”  Alexandre M. Samoylov, Tatiana I. Samoylova, Mark G. Hartell, Suram T. Pathirana, Bruce F. Smith, and Vitaly J. Vodyanoy.  Biomolecular Engineering 18:269-272, 2002. “Targeting peptides for microglia identified via phage display.”  T.I. Samoylova, B.Y. Ahmed, V. Vodyanoy, N.E. Morrison, A.M. Samoylov, L.P. Globa, H.J. Baker, and N.R. Cox.  NeuroImmunology 127:13-21, 2002. “Member of the ampakine class of memory enhancers prologs the single channel open times of reconstituted AMPA receptors."  V. Suppiramaniam, B.A. Bahr, S. Sinnarajah, K. Owens, G. Rogers, S.Yilma, and V. Vodyanoy. Synapse 40:154-158, 2001. “RGS2 ihibits Gsa sgnaling by ipairing activation of type III, V, and VI adenylyl cyclases.”  S. Sinnarajah, C.W. Dessauer, D. Srikumar, J. Chen, J. Yuen, J. Dennis, S. Yilma, E. E. Morrison, V. Vodyanoy and J. H. Kehrl.  Nature 409:1051-1055, 2001. “Rapid and sensitive biosensor for Salmonella.”  S.T. Pathirana, J. Barbaree, B.A. Chin, M.G. Hartell, W.C. Neely, and V. Vodyanoy.  Biosensors & Bioelectronics 15:135-141, 2000. "Heparin modulates the single channel kinetics of reconstituted AMPA receptors from rat brain." Srikumar Sinnarajah, Vishnu Suppiramaniam, Kolappa Prem Kumar, Randy A. Hall, Ben A. Bahr, and Vitaly Vodyanoy. Synapse 31:203-209, 1999. “Inhibition and enhancement of odorants-induced cAMP accumulation in rat olfactory cilia by antibodies directed against Gai-protein subunits.”  S. Sinnarajah, P.I. Ezeh,  S. Pathirana, A.G. Moss, E.E. Morrison, and V. Vodyanoy. FEBS Letters 426:377-380, 1998. "Molecular sensor based on olfactory transduction" in: Molecular Electronics: Biosensors and Biocomputers. Felix T. Hong (Ed.).  Plenum Publishing Corp., New York, pp. 319-345, 1989. "Condensing and expanding effects of the odorants (+)-and (-)-carvone on phospholipid monolayers".  S. Pathirana, W.C. Neely, and V. Vodyanoy.  Langmuir 14:679-682, 1998. "Efects of heparin on the properties of solubilized and reconstituted rat brain AMPA receptors".  Randy A. Hall, Vitaly Vodyanoy, Alex Quan, Srikumar Sinnarajah, Vishnu Suppiramaniam, Markus Kessler, and Ben A. Bahr. Neuroscience Letters 217:179-183, 1996. Assembly of cadmium stearate and valinomycin molecules assists complexing of K+ in mixed Langmuir-Blodgett films.  S. Pathirana, L.J. Myers, V. Vodyanoy, and W.C. Neely.  Supramolecular Science 3:149-154, 1996. "Stearic acid assisted complexation of K+ by valinomycin in monolayers."  V. Vodyanoy, S. Pathirana, and W.C. Neely.  Langmuir 10:1354-1357, 1994. "Single channel recordings of reconstituted AMPA receptors reveal low and high conductance states."  V. Vodyanoy, B.A. Bahr, V. Suppiramaniam, R.A. Hall, M. Baudry and G. Lynch.  Neuroscience Letters 150:80-84, 1993. "Functional reconstitution of a-amino-3-hydroxy-5-methylisoxazole-4-propanate (AMPA) receptors from rat brain."  B.A. Bahr, V. Vodyanoy, R.A. Hall, V. Suppiramaniam, M. Kessler, K. Sumikawa, and G. Lynch.  J. Neurochemistry 59:1979-1982, 1992. "Chiral recognition of odorants (+)- and (-)-carvone by phospholipid monolayers."  S. Pathirana, W.C. Neely, L.J. Myers, and V. Vodyanoy.  J. Am. Chem. Soc. 114:1404-1405, 1992. "Interaction of valinomycin and stearic acid in monolayers."  S. Pathirana, W.C. Neely, L.J. Myers, and V. Vodyanoy.  Langmuir 8:1984-1987, 1992. "Glutamate-activated ion channels."  V. Vodyanoy.  CNS Neurotransmitters and Neuromodulators, Vol 1, CRC press, pp. 127-142, 1995. "Cyclic nucleotide-gated electrical activity in olfactory receptors".  V. Vodyanoy, in: Receptor and Transduction Mechanisms in Taste and Olfaction.  Joseph G. Brand and John H. Teeter (Eds.).  Marcel Dekker, New York, pp. 319-345, 1989. "Molecular sensor based on olfactory transduction" in: Molecular Electronics: Biosensors and Biocomputers. Felix T. Hong (Ed.).  Plenum Publishing Corp., New York, pp. 319-345, 1989. For a complete list of publications, visit https://fp.auburn.edu/vetmed/vvgroup/vv/articles.aspx [back to top]