School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH What Can We See With Quantum Dots (Q-Dots)? Faculty: Elisabeth Papazoglou, PhD, Drexel University. Relative Size of Q-Dots PROJECTOVERVIEWPROJECTOVERVIEW

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Faculty: Elisabeth Papazoglou, PhD, Drexel University. Absorption and Emission Spectra PROJECTOVERVIEWPROJECTOVERVIEW

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Valuable model system due to fluorescence – can guide the understanding of other nanosystems. Two types of cell cultures: 1. Bovine aortic endothelial cells (BAEC). 2. Ductal carcinoma cells – cancer cells. Plate-coated with Q-dot solution. With or without collagen – culture cells. Trypsinize – centrifuge. Fluorescence Microscopy – to see if the cells did in fact uptake the Q-dots. BAEC + Qdots – Lake Placid Blue – CT - Phase Contrast – 40X Gelatin coated 25 well plate – transferred cells-Nikon Inverted - FITCI BAEC + Qdots – Lake Placid Blue – CT - Fluorescence – 40X Gelatin coated 25 well plate – transferred cells-Nikon Inverted - FITCI Faculty: Elisabeth Papazoglou, PhD, Drexel University. How Do Particles of This Size Interact With Cells? PROJECTOVERVIEWPROJECTOVERVIEW

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Cancer Cells + Qdots – Lake Placid Blue – CT - Phase Contrast – 40X Gelatin coated 25 well plate – transferred cells-Nikon Inverted - FITCI Cancer Cells + Qdots – Lake Placid Blue – CT - Fluorescence – 40X Gelatin coated 25 well plate – transferred cells-Nikon Inverted - FITCI Faculty: Elisabeth Papazoglou, PhD, Drexel University. Current Activities Functionalization with Epidermal Growth Factor Raman Spectroscopy Near Future Diffusion through Skin Reconstruct Encapsulation in Liposomes Cancer Cells and Q-dots PROJECTOVERVIEWPROJECTOVERVIEW

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaie, PhD, Bahar Edrissi, Emir Rezvan, all at Drexel University. Q-dots As Probes of Cellular Pathways PROJECTOVERVIEWPROJECTOVERVIEW What are the Q-dots? The quantum dots (Evident Technologies) are semiconductor nanocrystals (diameter 2-10 nm) made from CdSe with an outside shell of ZnS. Their size and their surface chemistry are the main determinants of their optical, electronic, and chemical properties. We used Carboxyl and Amine terminated Qdots with diameters of nm. Preliminary Experiments Cell Types: Bovine Aortic Endothelial Cells (BAEC) and Human Ductile Carcinoma Cells (HDC) Q-dots: Lake Placid Blue (AT and CT terminated) Techniques: Syringe loading and Q-dot coating Live/dead assay: Calcein AM and Ethidium homodimer-1 Phase Contrast (40X)Fluorescence (40X) BAEC

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Syringe Loading Techique PROJECTOVERVIEWPROJECTOVERVIEW BAEC + AT + Pluronic F-68 (3 hrs) Syringe loading (40X) BAEC + AT + Pluronic F-68 (3 hrs) No Syringe loading (40X) No significant improvement in fluorescence. AT Versus CT Q-dots (Q-dot Coating Technique) AT Q-dots usually show a greater fluorescence compared to CT Q-dots. BAEC + AT (25 hrs) Magnification of 10X BAEC + CT (25 hrs) Magnification of 10X Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaie, PhD, Bahar Edrissi, Emir Rezvan, all at Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH BAEC Versus HDC (Q-dot Coating Technique) PROJECTOVERVIEWPROJECTOVERVIEW BAEC shows a more dispersed pattern of Q-dot uptake, whereas HDC shows a more aggregated one. BAEC + CT (48 hrs) Magnification of 40X HDC + CT (48 hrs) Magnification of 40X Live / Dead Assay (Q-dot Coating Technique) Q-dot contact for a short time period and the transfer process do not seem to cause significant damage to the cells. BAEC + AT (4 hrs) Live Assay (4X)BAEC + AT (4 hrs) Dead Assay (4X) Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaie, PhD, Bahar Edrissi, Emir Rezvan, all at Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Goals and Future Work PROJECTOVERVIEWPROJECTOVERVIEW Develop a model system for interaction of nanoparticles with cells. Explore the concept of non-invasive, reproducible introduction of nanoparticles into the cell. Explore the concept of non-invasive, reproducible introduction of nanoparticles into the cell. Develop a simple, robust technique for detection of multiple enzymes and proteins within a cell. Develop a simple, robust technique for detection of multiple enzymes and proteins within a cell. Functionalize Q-dots with growth factors. Functionalize Q-dots with growth factors.  Attach Q-dots to EGF.  Follow EGF pathway inside the cell. Attach Streptavidin Q-dots to biotinylated antibodies to simultaneously detect various proteins within a cell. Attach Streptavidin Q-dots to biotinylated antibodies to simultaneously detect various proteins within a cell.  For example, biotinylated anti-EGF and biotinylated anti-PERK. Attach antibody to Q-dots and compare with Streptavidin-biotin technique. Attach antibody to Q-dots and compare with Streptavidin-biotin technique. Devise technique to follow Q-dots' fate within the cell. Devise technique to follow Q-dots' fate within the cell. Key Challenges Acces to confocal microscope with UV excitation. Acces to confocal microscope with UV excitation. Optimization of the EGF attachment to Q-dots. Optimization of the EGF attachment to Q-dots. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaie, PhD, Bahar Edrissi, Emir Rezvan, all at Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Detecting Proteins With Light In Solution and In Cells PROJECTOVERVIEWPROJECTOVERVIEW YYYY Proteins are much smaller (~10nm) than one-half the wavelength of visible light (~200nm), which is the necessary size for direct imaging. Solution: Use light effects such as evanescent waves to induce amplification through surface plasmon resonance (SPR), fluorescence, and FRET. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Nanoprobe Operation PROJECTOVERVIEWPROJECTOVERVIEW Confine excitation to sub-wavelength aperture using multiple reflections (thick Ag film [100nm] for UV confinement). Use only evanescent excitation to reduce background signal and calculated focal volume. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Nanoprobe Setup PROJECTOVERVIEWPROJECTOVERVIEW UV light Needle-like fiber Collect/Count Photons Quantify Concentration Y <150 nm Cell under inverted microscope Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Nanoprobe Advantages PROJECTOVERVIEWPROJECTOVERVIEW Controlled light delivery to specified location Intra-membrane and intracellular measurements Reported M intracellular detection with immuno-based detection scheme Single photon resolution using PMT Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Antibody Functionalization of Silica Surface PROJECTOVERVIEWPROJECTOVERVIEW Phase contrast and fluorescent contrast images of control fiber (left) and FITC labeled antibody functionalized fiber (right). The fluorescence in the control is due to a defect in the fiber, as seen in the phase image. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Preliminary Cell Viability PROJECTOVERVIEWPROJECTOVERVIEW Trypan blue experiments with four probe diameters. After several cells were probed with the same tip, there was an observed residue buildup on the tip. After this event, subsequent probes resulted in membrane adherence to the probe, thereby rupturing cellular integrity. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Alternative Detection Schemes PROJECTOVERVIEWPROJECTOVERVIEW Is Fluorescent Labeling the only Option??? Alternative to labeling: Surfaced Enhanced Raman Scattering (SERS) nanoprobes. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Evanescent Waves Induce SPR PROJECTOVERVIEWPROJECTOVERVIEW TIR light at some critical angle can induce this collective electron oscillation through the coupling of an evanescent wave’s energy. Surface Plasmon Resonance (or Polariton) is a collective oscillation of the charge at the surface of a thin metal film. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Surface Enhanced Raman of Zinc Oxide Powder PROJECTOVERVIEWPROJECTOVERVIEW Raman signal is indicated in red in the inset. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH Surface Enhanced Raman of Zinc Oxide Powder PROJECTOVERVIEWPROJECTOVERVIEW Raman signal indicated in blue in the inset. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.

School of Biomedical Engineering, Science & Health Systems V 1.0 SD [040227] NANOSCALE IMAGING AND SENSING RESEARCH SurfaceEnhanced Raman for Phosphate Buffer Saline Solution PROJECTOVERVIEWPROJECTOVERVIEW Raman signal from the tip of the gold rod (in the absence of gold colloid) is indicated in red in the inset. Raman signal from the tip of the gold rod (in the presence of gold colloid) is indicated in blue in the inset. Faculty: Elisabeth Papazoglou, PhD, Kambiz Pourrezaei, PhD, Drexel University.