Live single cell imaging assays in glass microwells produced by laser-induced deep etching.

Miniaturization of cell culture substrates enables controlled analysis of living cells in confined micro-scale environments. This is particularly suitable for imaging individual cells over time, as they can be monitored without escaping the imaging field-of-view (FoV). Glass materials are ideal for most microscopy applications. However, with current methods used in life sciences, glass microfabrication is limited in terms of either freedom of design, quality, or throughput. In this work, we introduce laser-induced deep etching (LIDE) as a method for producing glass microwell arrays for live single cell imaging assays. We demonstrate novel microwell arrays with deep, high-aspect ratio wells that have rounded, dimpled or flat bottom profiles in either single-layer or double-layer glass chips. The microwells are evaluated for microscopy-based analysis of long-term cell culture, clonal expansion, laterally organized cell seeding, subcellular mechanics during migration and immune cell cytotoxicity assays of both adherent and suspension cells. It is shown that all types of microwells can support viable cell cultures and imaging with single cell resolution, and we highlight specific benefits of each microwell design for different applications. We believe that high-quality glass microwell arrays enabled by LIDE provide a great option for high-content and high-resolution imaging-based live cell assays with a broad range of potential applications within life sciences.

End-group functionalization of poly(3-hexylthiophene) as an efficient route to photosensitize nanocrystalline TiO2 films for photovoltaic applications.

Bulk heterojunction (BHJ) and dye-sensitized solar cells (DSSCs) have seen increased popularity over recent years and each technology has experienced tremendous improvements in power conversion efficiencies (PCEs), reaching 8 and 12%, respectively. The two technologies have been on independent improvement pathways, and this work establishes a link between them by using the archetypical hole conductor (poly-3-hexylthiophene, P3HT) in BHJs as a sensitizer on TiO(2) for DSSC applications. Three polymers were synthesized and examined as potential TiO(2) sensitizers in DSSCs under AM1.5 solar radiation. Using Grignard metathesis, regioregular P3HT was synthesized then functionalized with either one or two cyanoacrylic acid linker moieties to bind to the TiO(2) surface. End-group modification resulted in minimal changes to the optical and electronic properties as compared to pristine P3HT. Cyclic voltammetry (CV) experiments at anodic potentials of adsorbed sensitizer quantified the amount of alkylthiophene adsorbed on the TiO(2), whereas under reductive sweeps, cyanoacrylic acid end-group binding was determined. CVs of each polymer indicated that loading was drastically different as compared to pristine P3HT with the lowest loading on TiO(2) and monofunctionalized P3HT exhibited the highest loading. The DSSCs showed power conversion efficiencies (PCEs) of 0.1%, 0.2 and 2.2% for the polymer-sensitized TiO(2) of the unfunctionalized, monofunctionalized and difunctionalized polymers, respectively. DSSCs were then subjected to electrochemical impedance spectroscopy (EIS) in the dark and under monochromatic light radiation. The large variance in performance for the functionalized-P3HT sensitizers is attributed to differences in the adsorption modes of sensitizer on the TiO(2) surface, which in the difunctionalized case limits electrolyte recombination and favors forward charge transfer reactions.

Enzymatic ring opening of an iron corrole by plant-type heme oxygenases: unexpected substrate and protein selectivities.

Heme oxygenases are widely distributed enzymes involved in the oxidative cleavage of the heme macrocycle that yields the open-chain tetrapyrrole biliverdin IX, CO, and iron. For the first time, two regioisomeric iron corroles [α-CH- and γ-CH-Fe(cor)] have been utilized as artificial substrate and cofactor analogues to mammalian, plant, cyanobacterial, and bacterial heme oxygenases. The non-natural enzymatic cleavage of γ-CH-Fe(cor), catalyzed by plant-type heme oxygenases from Arabidopsis thaliana and Synechocystis sp., happens selectively at the unexpected bipyrrolic position and yields a biomimetic biliverdin-like product. The reaction is selective for this corrole regioisomer and for plant-type heme oxygenases and is the first report of an enzymatic corrole ring opening.

Metal-rich organometallics.

The scope of organometallics has recently been expanded considerably, when researchers have started to employ these compounds in the context of materials chemistry. Traditionally, organometallic complexes have been and continue to be important tools for many (catalytic) chemical transformations. This perspective highlights how metal-rich organometallics, that is compounds with more than one type of metal, can play an important role in the advancement of new value-added functional materials.