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Molecular beacon fluorescence detection of pseudogene SHMT-psI using two molecular beacon probes, E8/E10 and E9/E10 Evanescent Wave Sensing Detection of Phosphorylated STAT3 Protein by Fiber-Based Sensor Fluorescence Generated at Surface Exciting Laser Cladding Core Fluorophore labeled Phospho-STAT3 antibodies. Fluorescence coupled into fiber. Phospho-STAT3 1500 Fluorescence intensity in a.u. Phospho-STAT3 antibodies (IgG) Evanascent wave excited Alexa-430 Spectra in two different probes. 1200 Signal from stimulated Cells Signal from non-stimulated Cells 900 600 300 0 400 STAT - Signal Transducer and Activator of Transcription protein 450 500 550 600 Wavelength in nm 650 700 Surface Plasmon Sensors Kretschmann (ATR) geometry used to excite surface plasmons Surface plasmon resonance curves. The left-hand side curve is for just the silver film (labeled Ag); the right-hand side shows the curve (labeled Ag/p-4-BCMU) shifted on the deposition of a monolayer Langmuir-Blodgett film of poly-4-BCMU on the silver film Photonic Crystal Sensors Close packing of colloidal nanospheres to form a photonic crystal of close-packed colloidal array. (Left) Atomic Force Microscope (AFM) image of the surface layer. (Right) Scanning Electron Microscope (SEM) image of a cross-section (Markowicz and Prasad; unpublished). Visible extinction spectra showing how diffraction depends on the glucose concentration for a 125µmthick PCCA glucose sensor. The ordinate is given as –log T, where T is the transmittance. The PCCA expands for concentrations between 0.1 and 0.5 mM glucose. FLOW CYTOMETRY Basic concepts: - Use molecularly specific fluorescent tags to label cells - Single-file flow of cells (hydrodynamic focusing) - Laser illumination of single cells - Multiparameter fluorescence detection -Data analyses and/or sorting ACIS, November 2003 Schematics of a five parameter flow cytometer showing det of collection scheme. Various parameters monitored ACIS, November 2003 Clusters of cells represent distinct populations of cells ACIS, November 2003 Applications • • • • • • HIV Monitoring Leukemia or Lymphoma Immunophenotyping Organ Transplant Monitoring DNA Analysis for Tumor Ploidy and SPF Primary and Secondary Immunodeficiency Bacterial contamination in food and water Biophotonics Opportunities • • • • • • • • • • • Modeling of light propagation in tissue In-vivo Bioimaging, Spectroscopy, and Optical Biopsy Real-Time monitoring of Drug Interactions Optical Biopsy Nano-Biophotonic Probes Multiphoton Processes for Biotechnology Femtolaser based biotechnology\ Nanoarray multianalytic sensors Multiparameter Molecular Flowcytometry Single Molecule Biofunctions Nanomedicine Acknowledgements Researchers at the Institute: Prof. E. Bergey Prof. A. Cartwright Prof. M. Swihart Prof. E. Furlani Dr. A. Kachynski Dr. A. Kuzmin Dr. Y. Sahoo Dr. H. Pudavar Dr. T. Ohulchanskyy Dr. D. Bharali Dr. D. Lucey Dr. K. Baba Dr. J. Liu Outside Collaborators Prof. R.Boyd Prof. J.Haus Prof. J M J Frechet Prof. M. Stachowiak Dr. A. Oseroff Dr. R. Pandey Dr. J. Morgan Dr. P Dandona DURINT/AFSOR Dr. Charles Lee “Lighting the Way to Technology through Innovation” The Institute for Lasers, Photonics and Biophotonics University at Buffalo Biophotonics P.N.Prasad www.biophotonics.buffalo.edu