Visualizing single-molecule diffusion in mesoporous materials.

Periodic mesoporous materials formed through the cooperative self-assembly of surfactants and framework building blocks can assume a variety of structures, and their widely tuneable properties make them attractive hosts for numerous applications. Because the molecular movement in the pore system is the most important and defining characteristic of porous materials, it is of interest to learn about this behaviour as a function of local structure. Generally, individual fluorescent dye molecules can be used as molecular beacons with which to explore the structure of--and the dynamics within--these porous hosts, and single-molecule fluorescence techniques provide detailed insights into the dynamics of various processes, ranging from biology to heterogeneous catalysis. However, optical microscopy methods cannot directly image the mesoporous structure of the host system accommodating the diffusing molecules, whereas transmission electron microscopy provides detailed images of the porous structure, but no dynamic information. It has therefore not been possible to 'see' how molecules diffuse in a real nanoscale pore structure. Here we present a combination of electron microscopic mapping and optical single-molecule tracking experiments to reveal how a single luminescent dye molecule travels through linear or strongly curved sections of a mesoporous channel system. In our approach we directly correlate porous structures detected by transmission electron microscopy with the diffusion dynamics of single molecules detected by optical microscopy. This opens up new ways of understanding the interactions of host and guest.

Fabrication of metallized nanopores in silicon nitride membranes for single-molecule sensing.

The fabrication and characterization of a metallized nanopore structure for the sensing of single molecules is described. Pores of varying diameters (>10 nm) are patterned into free-standing silicon nitride membranes by electron-beam lithography and reactive ion etching. Structural characterization by transmission electron microscopy (TEM) and tomography reveals a conical pore shape with a 40 degrees aperture. Metal films of Ti/Au are vapor deposited and the pore shape and shrinking are studied as a function of evaporated film thickness. TEM tomography analysis confirms metalization of the inner pore walls as well as conservation of the conical pore shape. In electrical measurements of the transpore current in aqueous electrolyte solution, the pores feature very low noise. The applicability of the metallized pores for stochastic sensing is demonstrated in real-time translocation experiments of single lambda-DNA molecules. We observe exceptionally long-lasting current blockades with a fine structure of distinct current levels, suggesting an attractive interaction between the DNA and the PEGylated metallic pore walls.

Multiple core-shell functionalized colloidal mesoporous silica nanoparticles.

The selective functionalization of the inner and outer surfaces of colloidal mesoporous silica (CMS) nanoparticles with different trialkoxysilanes, following a newly developed delayed co-condensation approach, results in bifunctional CMS. Complementary CMS nanoparticles were prepared with two different functional groups located either on the outer shell or in the inner core of the particle. The identification and localization of the functional groups was achieved by means of different techniques including zeta potential, nitrogen sorption measurements, and fluorescence spectroscopy. This last technique was applied to fluorescein isothiocyanate (FITC)-labeled CMS featuring aminopropyl functional groups on the periphery or the internal pore surface of the particles. Fluorescence quenching experiments were carried out with dodecanethiolate-stabilized gold nanoparticles having a diameter greater than the pore size of the CMS. It could be shown that fluorescence quenching occurs only when the FITC is positioned on the outer surface of the CMS nanoparticles, whereas no quenching was observed for FITC located in the inner core of the nanoparticle. These results clearly confirm the controlled localization of the aminopropyl groups in the nanometer space of the CMS particles. Our approach thus offers the opportunity to synthesize, in a novel multistep co-condensation strategy, various bifunctional mesoporous nanoparticles with controlled localization of different functional groups in the inner core or on the outer shell of the nanoparticle.

Discrete tomography of demanding samples based on a modified SIRT algorithm.

The 3D structure of three particularly challenging samples was reconstructed by electron tomography. Due to sample limitations resulting in a large missing wedge and large tilt increments respectively the 3D structure could not be reconstructed by standard iterative algorithms; even a recently developed discrete algorithm failed until the input parameters for discrete reconstruction were improved. These challenges were addressed by adding a mask in each step of the preceding standard iterative reconstruction, setting all voxels known to be vacuum as zero, thus improving the segmentation and the 3D starting model. The position of these vacuum voxels is obtained from TEM images or other measurement data.