Replacing a Century Old Technique - Modern Spectroscopy Can Supplant Gram Staining.

Rapid and accurate Gram differentiation is paramount as the first step of pathogen identification and antibiotics administration. However, the current method requires additional reagents, is time-consuming, and is operator dependent. Here we show the principle of tip enhanced Raman spectroscopy (TERS) can differentiate between Gram negative and positive species, by detecting the changes in tip-enhancement in the Raman scattering from the bacteria's lipid-bilayer membrane, which specifically enhances Gram negative bacteria.

Deposition and Characterization of Roughened Surfaces.

Phase separation occurs whenever a solvent leaves a solution of strongly incompatible polymers. This can happen in bulk and in films. Films can be tailored as substrates for multiple applications such as solar cells, surface catalysis, and antireflection coatings. In this study, polystyrene (PS) was dissolved with polyvinyl acetate (PVAc) in different ratios using chlorobenzene as the solvent. Thin films of different ratios of PS and PVAc were deposited on glass via spin coating. The deposited films were investigated for their morphology, strain, surface area, and Raman scattering. The incompatibility between the two polymers leads to the growth of roughened PVAc islands supported by the PS matrix. A down shift in the Raman PVAc signal was observed in the combined film as compared with a 100% PVAc film, which was attributed to the high strain of PVAc that grew as tips. As the PVAc concentration in the polymer blend increases, the porous regions in the film expand and the amount and height of PVAc tips increase as well, up to the point where the pores merge to create a uniform surface. The optimal ratio for the deposition of a uniformly roughened surface is 75% PVAc and 25% PS. For demonstrating a possible application, we applied the partially roughened surface as a substrate for surface-enhanced Raman scattering and demonstrated at least 500% increase in the signal intensity measured in roughened areas. This is explained by the rod effect from the PVAc tips.

Exploring chromatin organization mechanisms through its dynamic properties.

The organization of the genome in the nucleus is believed to be crucial for different cellular functions. It is known that chromosomes fold into distinct territories, but little is known about the mechanisms that maintain these territories. To explore these mechanisms, we used various live-cell imaging methods, including single particle tracking to characterize the diffusion properties of different genomic regions in live cells. Chromatin diffusion is found to be slow and anomalous; in vast contrast, depletion of lamin A protein significantly increases chromatin motion, and the diffusion pattern of chromatin transforms from slow anomalous to fast normal. More than this, depletion of lamin A protein also affects the dynamics of nuclear bodies. Our findings indicate that chromatin motion is mediated by lamin A and we suggest that constrained chromatin mobility allows to maintain chromosome territories. Thus, the discovery of this function of nucleoplasmic lamin A proteins sheds light on the maintenance mechanism of chromosome territories in the interphase nucleus, which ensures the proper function of the genome.