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.
The short-term goal of this project is development of new strategies, methodologies and sensor platforms for rapid, real-time and label-free detection of Staphylococcus aureus and MRSA. The long-term objectives include development of universal sensing technology for the detection a variety of target biothreat agents. This project is designed to demonstrate feasibility of proposed concepts and approaches by accomplishing four specific aims. Our goal is to design a target-responsive system that can detect and quantitatively measure multiple biothreat agents based on highly specific biomolecular recognition. The operational principle of such a system is based on the specific recognition of target agent using highly specific lytic phage, and further development of analyte-specific output signal using several efficient and highly sensitive label-free approaches such as surface plasmon resonance (SPR) spectroscopy and grating-based light diffraction. While the current effort will be focused on the detection of S. aureus, any developed approaches and technologies could be universally applicable using different biorecognition elements (phages and antibodies) capable of recognizing a large variety of target agents. Collaboration with Dr. Sorokulova, B. Chin, B. Wilamowski. Supported by AUDFS USDA funds.
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).
The super resolution of the optical system is accomplished by two special features of the system. First, the effect of the annular illumination is the narrowing of the central spot of the diffraction pattern and increase of the intensity of the diffraction fringes. A similar effect of annular aperture is known and used in telescopes for the resolving power increase. Second, the illumination system produces a coherent illumination of small (compared to wavelength) object and thus the coherent image. Hence, partial cancellation of the diffraction amplitude occurs when the antiphase subsequent fringes of diffraction are added in the image. This property makes the image edge sharper and thus increases the resolution. The structural annual illumination gives an additional advantage to this system in that provides the capability to produce optical sectioning of thin samples similar to that found in confocal microscopy. The optical sectioning allows for the discerning of in-focus image from out-of-focus structures. It allows the user to see not only the sample cutting surface, but also makes it possible to glance inside optical sections below the cutting plane. Fine focusing and placement of the focus in any depth of the sample allows for positioning of the focus at a desirable increment. In this manner, a three-dimensional profile of the sample can be obtained. The system was successfully tested for observation of still and motile cells, nanoforms and vesicles produced by fragmented erythrocytes [81-83], and by characterization of primo nodes and vessels.
Primo Vascular System:
In the beginning of the 1960s, a North Korean scientist, Bong Han Kim, a professor of Pyongyang Medical College, discovered that the acupuncture meridian system was represented by a system of microcapillaries (Bonghan ducts). Kim Bong Han’s studies suggest that the meridian system is actually a physical microtubular network. This microcapillaries serve as special ducts in which a certain liquid. Due to political reasons Dr. Kim was abruptly removed from his position in 1965 and no published reports by Dr. Kim appeared after that. His work was abandoned and forgotten. This microcapillary network is found throughout the body’s organs and in several areas it resides within close proximity to the skin. The network is actually a vascular system that contains a fluid comprised of a number of components thought to be important to maintain and restoring health. Due to its relative location to the skin and fascia, this system may lie at the heart of OMM techniques and may serve to explain some of the restorative effects produced by these procedures. Obviously, the present work will attempt to more fully characterize this system, its composition and its function and how it is involved in health maintenance and its susceptibility to OMM techniques. The system recently was renamed as primo vascular system. Collaboration with I. Sorokulova.
The objective of the present study is to characterize the initial chemoreceptive events in the olfactory system of the rat and to investigate how is odorant information translated into a functional electrical event in the olfactory bipolar cell. The directions of the study are as follows:
- Odorant responses of olfactory neurons (EOG, Whole Cell Patch Clamp).
- Chemosensitive ion channels.
- Study of the initial chemoreceptive events.
- Role of metal nanoparticles in olfaction.
- Development of an experimental prototype of an analytical chemosensitive device.
- Canine olfactory system. In collaboration with Dr. E. Morrison and I. Sorokulova. Supported by Fetzer Foundation and HDS.
Biopolymer for Preservation Without Refrigeration:
The water-soluble biopolymer is used to immobilize the biological material by replacing the water molecules with the molecules of plastic. The plastic can be cured without the raising of temperature or applying UV radiation. The cured plastic makes a solid protective film, stable to many organic solvents, but removable by the water solution. The method is based on the simple, single compound, and a single procedure process. The developed process of immobilization does not include any additives, accelerators, or plastifiers, nor involve high temperature or radiation to promote polymerization. The objectives are to characterize preservation of bacterial and viral materials in polymer and to determine effects of environmental factors on the preservation of bacteria and viruses in the polymer. Collaboration with Dr. I. Sorokulova. Supported by AUDFS USDA funds.
Small Protein Particles (Proteons) From Blood:
We have developed a method of fast proliferation of mall protein particles found in blood. The method is based on in vitro seeded polymerization and aggregation of proteins in plasma in sterile conditions at elevated temperatures and pressures. This method allows assembling (and disassembling) of misfolded proteins, in very large quantities, within an hour. The amplified proteins can be identified by known immunochemical methods. Our long-term goal is to develop a method of amplification and control of misfolded protein proliferation for diagnosis and treatment of prion-related and other conformational disorders.Specific aims: (1) Determine conditions controlling assembling and disassembling of protein particlesincluding temperature, pressure, other actors effecting stability of proteins in blood, natural and artificial seeds; chaotropic, chelating, nhibitory, and protective compounds. (2) Define and characterize properties of nuclei (seeds) participating in isfolding, polymerization, and aggregation of proteins. Supported by Fetzer Foundation.
- Compositions for and methods of controlling olfactory responses to odorants
- Methods for isolating proteons from plasma samples
- Compositions for and methods of controlling olfactory responses to odorants
- Processes for isolating proteon nucleation centers (PNCs) from a biological sample obtained from an animal
- Methods for detecting misfolded proteins in biological samples
- Magnetostrictive ligand sensor
- Simultaneous observation of darkfield images and fluorescence using filter and diaphragm
- Method of forming monolayers of phage-derived products and used thereof
- Use of pullulan to isolate and preserve biological material
- Microscope illumination device and adapter therefor
- High resolution optical microscope with cardioid condenser for brightfield and darkfield illumination
- Microscope illumination device and adapter
- Use of acacia gum to isolate and preserve a biological receptor on a biosensor
- Phage ligand sensor devices and uses thereof
- Method of isolation and self-assembly of small protein particles from blood and other biological materials
- Ligand sensor devices and uses thereof
- High-resolution optical microscope
- 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
- 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