3D printing is gaining traction in research and development as a way to quickly, cheaply, and easily manufacture polydimethylsiloxane (PDMS) molds. The most commonly used method is resin printing, which is relatively expensive and requires specialized printers. This study shows that polylactic acid (PLA) filament printing is a cheaper, more readily available alternative to resin printing, that does not inhibit the curing of PDMS. As a proof of concept, a PLA mold for PDMS-based wells was designed, and 3D printed. We introduce an effective method to smooth the printed PLA mold, based on chloroform vapor treatment. After this chemical post-processing step, the smoothened mold was used to cast a ring of PDMS prepolymer. The PDMS ring was attached to a glass coverslip after oxygen plasma treatment. The PDMS–glass well showed no leakage and was well suited to its intended use. When used for cell culturing, monocyte-derived dendritic cells (moDCs) showed no morphological anomalies, as tested by confocal microscopy, nor did they show an increase in cytokines, as tested using ELISA. This underlines the versatility and strength of PLA filament printing and exemplifies how it can be valuable to a researcher’s toolset.

The first step in infection of influenza A virus is contact with the host cell membrane, with which it later fuses. The composition of the target bilayer exerts a complex influence on both fusion efficiency and time. Here, an in vitro , single-particle approach is used to study this effect. Using total internal reflection fluorescence (TIRF) microscopy and a microfluidic flow cell, the hemifusion of single virions is visualized. Hemifusion efficiency and kinetics are studied while altering target bilayer cholesterol content and sialic-acid donor. Cholesterol ratios tested were 0%, 10%, 20%, and 40%. Sialic-acid donors GD1a and GYPA were used. Both cholesterol ratio and sialic-acid donors proved to have a significant effect on hemifusion efficiency. Furthermore, comparison between GD1a and GYPA conditions shows that the cholesterol dependence of the hemifusion time is severely affected by the sialic-acid donor. Only GD1a shows a clear increasing trend in hemifusion efficiency and time with increasing cholesterol concentration of the target bilayer with maximum rates for GD1A and 40% cholesterol. Overall our results show that sialic acid donor and target bilayer composition should be carefully chosen, depending on the desired hemifusion time and efficiency in the experiment.

Fusogenicity of CPE and CPK peptides in liposomes is, at least partially, dependent on the size of the particle. Under certain conditions (small liposomes), CPK alone is enough to trigger membrane fusion in both bulk and single-particle studies.

Microplastics have become ubiquitous in the environment and are increasingly found in a variety of matrices, including in animal feed. At the same time, the presence of plastics from packaging materials in animal feed is still prohibited in the European Union. This is a major barrier preventing certain organic waste streams from being repurposed as feed. However, issues associated with the presence of plastic particles in feed for ‘conventional’ livestock such as poultry and pigs (e.g. accumulation in stomach/other tissues) may not pose a problem to reared insects such as black soldier fly larvae (BSFL). In the hypothetical absence of such transfer, insects reared on feed contaminated with plastics could potentially be repurposed as feed. Such reuse would be a major opportunity for a more circular food chain. The aim of this exploratory study was to develop a method to determine the potential transfer of microplastics from the substrate to biomass of reared BSFL. Tested plastics consisted of low-density polyethylene (LDPE) of three different sizes, and polyethylene terephthalate (PET); to mimic the types of synthetic polymers commonly used for food packaging. The results showed no significant effects of dietary exposure of tested (micro-)plastics on larval yield and survival. We benchmarked fluorescence microscopy through Nile Red staining against a newly developed technique utilizing polarised light optical microscopy to identify plastic particles in BSFL. Tested LDPE particles were virtually absent from the post-exposure larvae. PET was recovered from the larvae, although it is unclear what the exact mechanism for this transfer was. More research is needed to validate these experimental findings with plastics of different types and particle sizes.