In-vivo two-photon imaging of the honey bee antennal lobe

Due to the honey bee's importance as a simple neural model, there is a great need for new functional imaging modalities. Herein we report on the use of two-photon microscopy for in-vivo functional and morphological imaging of the honey bee's olfactory system focusing on its primary centers, the antennal lobes (ALs). Our imaging platform allows for simultaneously obtaining both morphological measurements of the AL and in-vivo calcium recording of neural activities. By applying external odor stimuli to the bee's antennas, we were able to record the characteristic odor response maps. Compared to previous works where conventional fluorescence microscopy is used, our approach offers all the typical advantages of multi-photon imaging, providing substantial enhancement in both spatial and temporal resolutions while minimizing photo-damages and autofluorescence contribution with a four-fold improvement in the functional signal. Moreover, the multi-photon associated extended penetration depth allows for functional imaging within profound glomeruli.Comment: 3 pages, 3 figure

From Functional Group Ensembles to Single Molecules: Scanning Force Microscopy of Supramolecular and Polymeric Systems

Surfaces in supramolecular and polymeric systems were characterized by scanning force microscopy (SFM) using probe tips functionalized with self-assembled monolayers (SAMs). This approach allows one to control the forces between tip and surface by immobilizing suitable molecules, which expose selected functional groups, onto gold-coated probes. The objective of this Thesis work was the extension of SFM with SAM-functionalized probes (so-called "chemical force microscopy", CFM) to technologically relevant surfacetreated polymers and elastomers with the ultimate aim of laterally resolved detection of functional group distributions on a sub-100 nm level. In measurements of interaction forces between a few or even individual molecules in supramolecular systems the transition from studying continuum to studying non-continuum properties was also attempted. In situ measurements of reaction kinetics using "inverted CFM" on a scale of 10 - 100 molecules were achieved. Furthermore, interactions between ensembles of functional groups and individual molecules were studied and provided fundamental insight into functional group distributions on polymer surfaces as well as rupture forces of individual host-guest complexes

Labyrinthine Island Growth during Pd/Ru(0001) Heteroepitaxy

Using low energy electron microscopy we observe that Pd deposited on Ru only attaches to small sections of the atomic step edges surrounding Pd islands. This causes a novel epitaxial growth mode in which islands advance in a snakelike motion, giving rise to labyrinthine patterns. Based on density functional theory together with scanning tunneling microscopy and low energy electron microscopy we propose that this growth mode is caused by a surface alloy forming around growing islands. This alloy gradually reduces step attachment rates, resulting in an instability that favors adatom attachment at fast advancing step sections

Real-Time analysis and visualization for single-molecule based super-resolution microscopy

Accurate multidimensional localization of isolated fluorescent emitters is a time consuming process in single-molecule based super-resolution microscopy. We demonstrate a functional method for real-time reconstruction with automatic feedback control, without compromising the localization accuracy. Compatible with high frame rates of EM-CCD cameras, it relies on a wavelet segmentation algorithm, together with a mix of CPU/GPU implementation. A combination with Gaussian fitting allows direct access to 3D localization. Automatic feedback control ensures optimal molecule density throughout the acquisition process. With this method, we significantly improve the efficiency and feasibility of localization-based super-resolution microscopy

Common-path multimodal optical microscopy

We have developed a common-path multimodal optical microscopy system that is capable of using a single optical source and a single camera to image amplitude, phase, and fluorescence features of a biological specimen. This is achieved by varying either contrast enhancement filters at the Fourier plane and/or neutral density/fluorescence filters in front of the CCD camera. The feasibility of the technique is demonstrated by obtaining brightfield, fluorescence, phase-contrast, spatially filtered, brightfield + fluorescence, phase +fluorescence, and edge-enhanced+fluorescence images of the same Drosophila embryo without the need for image registration and fusion. This comprehensive microscope has the capability of providing both structural and functional information and may be used for applications such as studying live-cell dynamics and in high throughput microscopy and automated microscopy

Graphene on Rh(111): STM and AFM studies

The electronic and crystallographic structure of the graphene/Rh(111) moir\'e lattice is studied via combination of density-functional theory calculations and scanning tunneling and atomic force microscopy (STM and AFM). Whereas the principal contrast between hills and valleys observed in STM does not depend on the sign of applied bias voltage, the contrast in atomically resolved AFM images strongly depends on the frequency shift of the oscillating AFM tip. The obtained results demonstrate the perspectives of application atomic force microscopy/spectroscopy for the probing of the chemical contrast at the surface.Comment: manuscript and supplementary information; submitted to Appl. Phys. Lett. on 01.03.201