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Our Facilities We have modern laboratories which are equipped to meet the requirements of a wide range of experimental research. (But, we still lack a lot !!) - Polymer synthesis laboratory - Polymer characterization laboratory - Cell-culture laboratory - Imaging laboratory - Nanofiltration Our Research Topics Involving Nanotechnology • Biomaterials/tissue eng • Drug/DNA/RNA delivery • Nanomedicine • Biosensors • Biochips • Imaging • Biocatalyst Development • Bioseparations (with emphasis on Nanotechniques) Biomaterials & Tissue engineering Microporous Biodegradable Electruspun PCL Scaffolds Microporous Biodegradable PLLA-CL Scaffolds prepared in supercritical CO2 NANOFIBROUS MATERIALS By Electrospinning Electrospinning is a unique method that produces polymeric nanofibers with diameter in the range of nano to a few microns using electrically driven jet of polymer solutions/ melts Composites for Hard Tissue Repair Poly(DL-Lactide/ε-caprolactone) with nano-size TCP Aim: To prepare a hard H3PO4 tissue filling material that is biodegradable, easily Ca(OH)2 applied precipitate HEAT + NH4OH centrifugation Average particle size; Gel form: 300 nm Dry form: 10 nm TCP gel Dring and Sintering DRUG/DNA/RNA DELIVERY At 28ºC Gene Expression Efficiency: 50% Imaging at Nanoscale SCANNING PROBE MICROSCOPIES • • • • Scanning Tunnel Microscopy Atomic Force Microscopy SPR Microscopy Imaging Ellipsometry Scanning Tunnelling Microscopy STM Gerd Binnig & Heinrich Rohrer NOBEL PRIZE in 1986 Binning and Rohrer, IBM ONLY Conducting surfaces and researcher’s in Switzerland molecular monolayers described the STM in 1981. It is essentially a superfine stylus 5 µHigh resolution straightforward that sweeps over a surface like a blind person’s walking stick. The stylus is few atom widths away and it has a molecularly or perhaps atomically fine tip. Atomic Force Microscopy (AFM) The AFM head employs an optical detection system in which the tip is attached to the underside of a reflective cantilever. A diode laser is focused onto the back of a reflective cantilever. As the tip scans the surface of the sample, the laser beam is deflected off the attached cantilever. And the photodetector measures the difference in light intensities between the upper and lower photodetectors, and then converts to voltage. Contact mode ATOMIC FORCE MICROSCOPY (AFM) ATOMIC FORCE MICROSCOPY (AFM) Erythrocytes DNA Antibody Cromosome SPR Microscope SPR CCD Camera L4 PC/Framegrabber Stages Stage Controller 2Θ (%) Flow Cell 100 Θ L3 Prism Filter L2 L1 Polarizer l/4 Waveplate HeNe Laser, 1.5 mW Reflection Intensity Substrate 0 I II Angle of Monitoring Following: W. Knoll,Colloid Polym. Sci. 1988, 109, 244 Imaging Ellipsometry Dextran on silicone Polarizer Analyzer Objective Compensator Sample (e.g. film) Substrate Dye monolayer Patterned surface FORCE MEASUREMENTS by using Modified AFM Tips BIOSENSORS ANALYTE SIGNAL BIORECEPTOR TRANSDUCER Enzyme Antibody Receptor DNA Electrochemical Optical Piezoelectric The bioreceptor is a biomolecule that recognizes the target analyte. The transducer converts the recognition event into a measurable signal. QUARTZ CRYSTAL MICROBALANCE Quartz crystal plate Contact Au electrodes UNIVERSAL SENSORS Inc. ATOMIC FORCE MICROSCOPY As molecular sensors ATOMIC FORCE MICROSCOPY As molecular sensors Representative fluorescence microscopy images: (A) and (B) before ssDNA immobilization; (C) and (D) carrying the immobilized ligand ssDNA MAGNETICALLY LOADED NANOPARTICLES Nanoparticles take on special properties because of their small size. Nanoparticles can be use as carriers or labels with a variety of markers, transported through various media, and interfaced both in vivo and in vitro. Nanoparticle technology over the last decade plays an important role in the diagnosis and treatment of cancer. Biomedical Applications Early disease detection As reporter platforms Imaging As contrast enhancing agents Drug delivery As a vehicle MAGNETIC NANOPARTICLES Magnetic and Superparamagnetic Particles – Colloid size: 1-5 µm – Magnetite (Fe3O4), Maghemite (γ- Fe2O3) Colloidal Magnetic Particles – Colloid size: 50-500 nm – Ferrofluids BIOCHIPS Thousands of genes and their products (i.e., proteins) in a given living organism function in a complicated and orchestrated way that creates the mystery of life Traditional methods in molecular biology generally work on a "one gene in one experiment" basis, which means that the throughput is very limited and the "whole picture" of gene function is hard to obtain Biochips, or also so-called “micro arrays” are “Labs on Chips” and allows simultaneous analysis of multiple samples DNA-CHIPS PROTEIN-CHIPS SURFACE PLASMON RESONANCE Microarray-based studies of Protein-DNA Interactions Using SPR dsDNAs with Different sequences DNA binding proteins Stealth printhead/pin Substrate Substrate platform Microtiter plate • Protein binding shifts SPR minimum to a higher angle. • Adsorption and desorption monitored by tracking SPR angle changes. • SPR response can be converted to a surface coverage. Proteins in buffer dsDNA array Sensing surface % Reflectivity Vacuum manifold Water bath Sonicator Blotter Adsorption Adsorption Time Desorption Selection and characterization of peptides specific to Platinum surfaces by phage display and preparation of their light sensitive nano-hybrid forms Identification of peptide sequences specific to inorganics (Platinum metal) via phage display and physical and chemical characterization of selected peptides Preparation of nanostructures consisting of Pt/putative peptide/light sensitive molecule (azobenzene) Ligand for any target Light-sensitive molecule Peptide Pt Substrate N N N N 360 nm N N 470 nm Control of ligand-target interaction N N I. Peptide selection via phage display Characterization of selected peptides Statistical analysis Alignment (Clustal W) Charge, (http://us.expasy.org/tools/pi), Hydrophilicity Fluorescence microscopy Cross-specificity with powders and arrays Binding characteristics with QCM, SPR SD152 PTSTGQA b) Coupling of peptide to azobenzene molecule Characterization: Functional Grup analysis (FTIR, 1H-NMR, 13C- NMR) Ninhydrin assay, Absorption behaviour, Binding study with QCM ( ka, kd) Contact angle, elipsometer, AFM on Platinum surfaces -Δf (Hz) time (s) GENE THERAPY AIM: To avoid restenosis by Gene Therapy Proliferation of smooth muscle cells are stopped by blocking the MMP-2 enzyme. For this purpose, inhibitor gene is transfected to the smooth muscle cells. 100 µl (1mg/ml) polymer pH 5.95 10 g plasmid in NaCI 60x 103 smc/ well GENE EXPRESSION OF SMOOTH MUSCLE CELLS ea b f SMC’s transfected with Poly(NIPA-co-MAH)/plasmid DNA complex (e) light microscope image (f) Fluorescence microscope image.