Microfluidics: From Crystallization to Serial Time-Resolved Crystallography

Capturing protein structural dynamics in real-time has tremendous potential in elucidating biological functions and providing information for structure-based drug design. While time-resolved structure determination has long been considered inaccessible for a vast majority of protein targets, serial methods for crystallography have remarkable potential in facilitating such analyses. Here, we review the impact of microfluidic technologies on protein crystal growth and X-ray diffraction analysis. In particular, we focus on applications of microfluidics for use in serial crystallography experiments for the time-resolved determination of protein structural dynamics

Near-field interference map due to a dipolar emission near the edge of a monocrystalline gold platelet

Point source excitation and point detection in the near-field provides new perspective to study the near-field optical phenomena of plasmonic nanostructures. Using the automated dual-tip scanning near-field optical microscope (SNOM), we have measured the optical near-field response of a dipolar emission near the edge of a monocrystalline gold platelet. The image dipole method was used to analytically calculate the interference pattern due to surface plasmon polaritons excited at the position of aperture tip and those reflected from edges of the gold platelet. The near-field enhancement was observed on the edges of the gold platelet. Our results verify that automated dual-tip SNOM is an intriguing technique for quantum plasmonic studies where deterministic coupling of quantum emitters and the detection of the near-field enhancement are of great interest

RpkA, a Highly Conserved GPCR with a Lipid Kinase Domain, Has a Role in Phagocytosis and Anti-Bacterial Defense

RpkA (Receptor phosphatidylinositol kinase A) is an unusual seven-helix transmembrane protein of Dictyostelium discoideum with a G protein coupled receptor (GPCR) signature and a C-terminal lipid kinase domain (GPCR-PIPK) predicted as a phosphatidylinositol-4-phosphate 5-kinase. RpkA-homologs are present in all so far sequenced Dictyostelidae as well as in several other lower eukaryotes like the oomycete Phytophthora, and in the Legionella host Acanthamoeba castellani. Here we show by immunofluorescence that RpkA localizes to endosomal membranes and is specifically recruited to phagosomes. RpkA interacts with the phagosomal protein complex V-ATPase as proteins of this complex co-precipitate with RpkA-GFP as well as with the GST-tagged PIPK domain of RpkA. Loss of RpkA leads to a defect in phagocytosis as measured by yeast particle uptake. The uptake of the pathogenic bacterium Legionella pneumophila was however unaltered whereas its intra-cellular replication was significantly enhanced in rpkA-. The difference between wild type and rpkA- was even more prominent when L. hackeliae was used. When we investigated the reason for the enhanced susceptibility for L. pneumophila of rpkA- we could not detect a difference in endosomal pH but rpkA- showed depletion of phosphoinositides (PIP and PIP2) when we compared metabolically labeled phosphoinositides from wild type and rpkA-. Furthermore rpkA- exhibited reduced nitrogen starvation tolerance, an indicator for a reduced autophagy rate. Our results indicate that RpkA is a component of the defense system of D. discoideum as well as other lower eukaryotes

Learning at large conferences:from the 'sage on the stage' to contemporary models of learning

AimTo explore and evaluate the affordances of a flipped classroom model applied to a research paper session within the professional development opportunity of a large conference setting.MethodAuthors were invited to present their research papers in a flipped classroom presentation format at two large, multi-national conferences. Before the session, authors and moderators met online to clarify features of the session, and preparation of the material. The research material was then posted online before the conference, to allow access by meeting attendees. During the sessions, moderators encouraged the audience to actively participate. An evaluation form was collected from the audience at the end of each session.ResultsParticipants found the session valuable, and appreciated the opportunity to engage in a meaningful dialogue with colleagues. However, the majority of the audience did not access the materials in advance. Lack of time, or technology-related issues were mentioned as potential challenges to such format.ConclusionIn the context of a large conference, a 'flipped session' format can facilitate active learning and a participatory culture of inquiry. However, to change the nature of how individuals learn collaboratively at large conferences means a change in the culture of continuous professional learning