They are employed in applications to study receptor-ligand binding in plate-based assays 2, single-molecule fluorescent microscopy analyses 3, and flow cytometry 4, as well as in applications requiring cell-type specific targeting such as theranostics 5, 6, 7 and in vivo imaging 8, 9. This protocol for preparation of nanoparticle probes relies solely on commercially available reagents and common equipment, breaking down the barriers to use nanoparticles in biological experiments.įluorescently labeled antibody and aptamer probes are used to visualize and quantify biological molecules in the fields of biology, chemistry, and biomedicine 1. Nanoparticles functionalized with DNA aptamers targeting histidine tags and VEGF protein had high affinity (EC 50s ranging from 3 to 12 nM) and specificity, and were more stable than conventional labels. A conjugation-annealing handle and DNA aptamer probe were attached to the azide-PEG nanoparticle surface either through reaction of pre-annealed handle and probe or through a stepwise reaction of the nanoparticles with the handle followed by aptamer annealing. Particle passivation was achieved by covalent attachment of amine-PEG-azide to carboxylated particles, neutralizing the surface charge from − 43 to − 15 mV. Fluorescent polystyrene nanoparticles, Thermo Fisher Scientific FluoSpheres, were used in these proof-of-principle studies. This work introduces a straightforward approach for preparation of fluorescent nanoparticle probes using commercially available reagents and common laboratory equipment. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their enhanced brightness, stability and multivalency however, challenges in functionalization and characterization have impeded their use. He is Product Manager for Malvern Panalytical light scattering and micro-calorimetry platforms and Pall Fortébio product lines.Fluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. He works for Alfatest since 2015, focusing on the molecular interactions and nanoparticle/protein characterization instrumentation. Andrea Pigozzo is graduated in Environmental Sciences at University Ca’ Foscari of Venice. New Zetasizer ULTRA live demonstration (replicate)Īndrea Pigozzo - Alfatest srl – Distributore Malvern Panalytical per l’Italiaĭr. See the whole nanoparticle sample with MADLS ®: particle characterization you can trust and easy, calibration-free number concentration measurements (replicate) Reliable DLS size data quicker with Adaptive Correlation (replicate) See the whole nanoparticle sample with MADLS ®: particle characterization you can trust and easy, calibration-free number concentration measurements Reliable DLS size data quicker with Adaptive Correlation
PROGRAMME: 2 replicate sessions at 9.15 and 11.00 The new Zetasizer ULTRA will be presented on site during a live demonstration. The workshop will be dedicated both to measurement principle and applicative examples to illustrate the benefits of Adaptive correlation and MADLS ® respect to classical DLS. The brand new Zetasizer ULTRA from Malvern Panalytical featuring these 2 techniques will be officially presented for the first time in Italy during this worshop. The workshop will be dedicated to the brand new and revolutionary Adaptive correlation and multi-angle Dynamic Light scattering MADLS ® techniques for more complete particle size distributions and calibration-free concentration analysis. Light scattering is a primary technique for this purpose, and Malvern Panalytical is the leader provider of light scattering instruments for nanoparticle characterization. The ultimate technology for nanoparticle size and number concentration analysisįaculty of Civil and Industrial EngineeringĬharacterizing size and concentration is a key step in the study and development of dispersed nanosystems and for the control of the final product properties and quality.