This PDF file contains the front matter associated with SPIE Proceedings Volume 8815, including the Title Page, Copyright Information, Table of Contents, and the Conference Committee listing.

Radiation-damaged nanodiamonds (NDs) are ideal optical contrast agents for photoacoustic (PA) imaging in biological tissues due to their good biocompatibility and high optical absorbance in the near-infrared (NIR) range. Acid treated NDs are oxidized to form carboxyl groups on the surface, functionalized with polyethylene glycol (PEG) and human epidermal growth factor receptor 2 (HER2) targeting ligand for breast cancer tumor imaging. Because of the specific binding of the ligand conjugated NDs to the HER2-overexpressing murine breast cancer cells (4T1.2 neu), the tumor tissues are significantly delineated from the surrounding normal tissue at wavelength of 820 nm under the PA imaging modality. Moreover, HER2 targeted NDs (HER2-PEG-NDs) result in higher accumulation in HER2 positive breast tumors as compared to non-targeted NDs after intravenous injection (i.v.). Longer retention time of HER-PEG-NDs is observed in HER2 overexpressing tumor model than that in negative tumor model (4T1.2). This demonstrates that targeting moiety conjugated NDs have great potential for the sensitive detection of cancer tumors and provide an attractive delivery strategy for anti-cancer drugs.

Super-resolution microscopy often employs asynchronous localizations of many single fluorescent emitters achieving resolution below the diffraction limit. This family of techniques typically uses statistical switching of emitters between dark and bright fluorescent states. Here we investigate how imaging repeated activations cycles of the same emitter influences the achieved image resolution. Furthermore, we ask the questions how long such a typical bright emitting state should be and is there an optimal number of switching events if the measurement time is fixed. We find that longer measurement times and hereby imaging more activation cycles is always beneficial for the attained image resolution. In the case of a fixed measurement time it turns out that there is a trade-off between the number of cycles and the product of localization density and uncertainty.

The precision in localizing a molecule is ultimately determined by the number of detected photons, which is in turn limited by photobleaching. Currently, fluorophores can be routinely localized to a few tens of nanometers at room temperature. In this work we demonstrate localization precision better than 3 Angstrom by substantial improvement of the molecular photostability at cryogenic temperatures. We discuss the challenges, solutions and promise of our methodology for high-performance co-localization and super-resolution microscopy.

Super-resolution microscopy has emerged as a powerful and non-invasive tool for the study of molecular processes both in vitro, but also as they occur in live cells. Here we present the application of direct stochastic optical reconstruction microscopy (dSTORM), a super-resolution technique based on single molecule localization, to determine the morphology of protein aggregates and of small extra- and intracellular structures. The technique reveals details down to 20 nm providing information on scales much smaller than the wavelength of the probing light. We use dSTORM in the study of amyloid fibril self-assembly processes associated with neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases. We show that the aggregation process can be followed kinetically and observe the emergence of amyloid structures in time as they occur in vitro. As an all optical technique, there is translation potential from studies in vitro to in vivo applications.

This study reports on the use of surface enhanced Raman scattering (SERS) as a non-destructive tool for detection and localisation of Porphyrin-Gold nanoparticles (GNP) conjugates at the subcellular level. Conjugates of the hydrophobic photosensitizer meso-Tetraphenylporphyrin (TPP) and GNPs were synthesized. The TPP-GNPs were characterized by by ultraviolet—visible absorption spectroscopy, fluorescence spectroscopy and transmission electron microscopy. TPPGNPs with a mean diameter of 12 nm were introduced into SW480 human colon adenocarcinoma cells. Single point SERS was applied in conjunction with fluorescence microscopy to localize the exogenous materials within the cells. Our results indicate that the TPP-GNP nanomaterials are distributed within cells in the cytoplasm. Overall our results indicate that Raman spectroscopy has the potential to be a high-throughput tool to localise nanoparticles in the subcellular environment.

Tip-enhanced Raman spectroscopy (TERS) offers one of the best techniques for analysis and imaging of molecule structures at nanoscale spatial resolution. An important issue in TERS is to improve the detection sensitivity of inherently weak Raman scattering so as to observe varieties of materials. For enhancement of the Raman signal, fully metallized tips are utilized in TERS, which enhance signals through plasmon oscillation at the tip apex. However, length of metal along the tip axis is on the order of a few to a few tens of micrometers, which means the plasmon resonant wavelength is much longer than the wavelength of the visible light used in TERS. From that point, if the tip has a metallic nanostructure on the apex, it would give better enhancement in the visible range compared with fully metallized tips. In this research, we employed photoreduction as a new fabrication method to grow a metallic nanostructure at the tip apex. We found a particular property of photoreduction that it occurs selectively at sharp corners, such as the tip apex of silicon cantilevers. Through this property, we succeeded in growing silver nanoparticles selectively at the tip apex. One of the advantages of the photoreduction is that the size of metal nanostructures is well controlled by optimizing various parameters. We controlled the size of silver nanoparticles from 100 to 400 nm by changing the laser exposure time. Furthermore, we obtained an order of magnitude higher enhancement from our fabricated tip compared with fully metalized tips through TERS measurements.

We have developed a NSOM technique that can map both the near optical field and the optical force using an atomic force microscope. This technique could be very useful for characterizing MEMs/NEMs devices, plasmonic nanoantennas, nano-photonic devices and biologically active substrates. Unlike conventional NSOM techniques that rely on an aperture fabricated on the end of an AFM tip to collect the optical signal this apertureless technique uses a lockin amplifier locked to the AFM tip vibrational frequency, to correlate the amplitude modulation of the back reflected optical signal to the strength of the optical field. And since we are not limited by the fabrication of an aperture the spatial resolution of the map is limited only by the size of a sharp AFM tip which for metallic coated tips can have a radius of curvature of 10 to 20 nm. For optical force mapping the incident laser is modulated and the lock-in amplifier is used to correlate the amplitude modulation of the vibrating AFM tip to strength of the optical gradient force. And in this way one can get a very accurate mapping of both the optical force and the optical field for any substrate of interest as long as it can be back illuminated. Lastly with an electrically monolithic substrate it is possible to correlate the amplitude modulation of the tunneling current to the optical field and obtain a spatial mapping that has a resolution of an STM, about 1 nm or maybe less.

Apertureless scattering-type Scanning Near-field Optical Microscopy (s-SNOM) has been used to study the electromagnetic response of infrared antennas below the diffraction limit. The ability to simultaneously resolve the phase and amplitude of the evanescent field relies on the implementation of several experimentally established background suppression techniques. We model the interaction of the probe with a patch antenna using the Finite Element Method (FEM). Green's theorem is used to predict the far-field, cross-polarized scattering and to construct the homodyne amplified signal. This approach allows study of important experimental phenomena, specifically the effects of the reference strength, demodulation harmonic, and detector location.

To establish an efficient way to utilize a gap mode plasmon in flocculates of MNPs, under external and ATR configurations, we controlled interaction between adsorbed species and metal nanostructures. We have successfully formed flocculates of AgNPs using electrostatic interaction between dissociated PMBA (-COO^-), protonated PATP (-NH³⁺) and counter ions (Mⁿ⁺, X^-), as well as van der Waals force between neutral PMBAs (-COOH) and PATP (-NH₂) on AgNPs. Detailed adsorbed state of PMBA and PATP as well as trapped counter ions were characterized using enormous SERS enhancement in flocculation-SERS. In a gap mode under an external geometry, most of thiol molecules on Ag films immobilized AgNPs through van der Waals force and electrostatic interaction. They showed similar Raman enhancement of 10⁸-10⁹, in accordance with those predicted by FDTD calculations. Only thiols with tert-methyl group did not immobilize any AgNPs due to steric hindrance. In a gap mode under ATR configuration, additional enhancement was obtained by a coupling of PSP and a gap mode.