Simultaneous Rayleigh/Mie and Raman/Fluorescence Characterization of Molecularly Functionalized Colloids by Correlative Single-Particle Real-Time Imaging in Suspension.

Many applications of nano- and microparticles require molecular functionalization. Assessing the heterogeneity of a colloidal sample in terms of its molecular functionalization is highly desirable but not accessible by conventional ensemble experiments. Retrieving this information necessitates single-particle experiments which simultaneously detect both functionalized and nonfunctionalized particles via two separate imaging channels. In this contribution, we present an optical setup for performing correlative single-particle imaging using laser light-sheet illumination: the first detection channel records elastic light scattering (Rayleigh/Mie), while the second channel detects inelastic light scattering (Raman) or fluorescence. The instrument is tested with Raman reporter-functionalized SERS-active metal nanoparticles (core/satellite silver nanoparticles, dimers and monomers of gold nanoparticles) and fluorophore-functionalized colloids (fluorescent polymer microparticles, dye-labeled protein on gold nanoparticles).

Extracellular functional noncoding nucleic acid bioaptamers and angiotropin RNP ribokines in vascularization and self-tolerance.

Endogenous extracellular and circulating functional small noncoding nucleic acids (ncNAs; <200 nucleotides) and complexes with proteins (ribonucleoproteins; RNPs) make up varying biolibraries of molecular imprints of cellular histories. They are nascently formed upon cellular activation by extrinsic (environmental) factors, including mitogens, cell-mediated immune memory reactions (Landsteiner-Chase-Lawrence transfer factors), and metabolic (hypoxia) and (physical) shear stress forces. Those factors are conventional models for epigenetic (non-Mendelian) vascular remodeling variations directed rather to proteinaceous gene expression and regulation than genomic DNA sequence changes. Structurally defined ncNAs are described as small hairpin nc-shRNA bioaptamers in interaction with proteins forming functional (Cu,Ca,Na,K)-metalloregulated complexes (CuRNP; angiotropins). As nonmitogenic angiomorphogen cytokines (ribokines), they may reprogram confluent quiescent (contact-inhibited) endothelial cell types to migratory, phagokinetically active phenotypes in the morphogenesis of tolerated neovascular patterns. Their functions in organized and mess-chaotic vascular patterns were investigated with regard to master gene, information, epigenetic, vascular, and tumor factors. Some ncNAs feature three-dimensional codes (3D episcripts) for distinct protein conformer phases. They are suggested as being specific recognition types, the estimated repertoires of which are superior in diversity and specificity to conventional immune (glyco-)proteins. For episcription of phenotype variations, they may address and integrate information flow on molecular shapes to protein-mediated nucleic acid processing and [post-]translational modification mechanisms in ncNA-, redox, and metalloregulated conformation phase pathway-locked loops (CPLL). Several vascular and cancer epigenetic regulator proteins are shown to be entangled by sharing helix-nucleating structural (proteomic) domains for interaction with functional nc-shRNA, termed K/RxxxH (K/R3H, -xK/RxxxHx(7-9)h/xx(7-9)h/xx(5-20)K/Rx-). This would suggest a tolerated mess-chaotic tumor vascularization as a bioaptamer disorder in ncNA-switched proteinaceous genetic and epigenetic processes.