EnderScope: a low-cost 3D printer-based scanning microscope for microplastic detection.

Low-cost and scalable technologies that allow people to measure microplastics in their local environment could facilitate a greater understanding of the global problem of marine microplastic pollution. A typical way to measure marine microplastic pollution involves imaging filtered seawater samples stained with a fluorescent dye to aid in the detection of microplastics. Although traditional fluorescence microscopy allows these particles to be manually counted and detected, this is a resource- and labour-intensive task. Here, we describe a novel, low-cost microscope for automated scanning and detection of microplastics in filtered seawater samples-the EnderScope. This microscope is based on the mechanics of a low-cost 3D printer (Creality Ender 3). The hotend of the printer is replaced with an optics module, allowing for the reliable and calibrated motion system of the 3D printer to be used for automated scanning over a large area (>20 × 20 cm). The EnderScope is capable of both reflected light and fluorescence imaging. In both configurations, we aimed to make the design as simple and cost-effective as possible, for example, by using low-cost LEDs for illumination and lighting gels as emission filters. We believe this tool is a cost-effective solution for microplastic measurement. This article is part of the Theo Murphy meeting issue 'Open, reproducible hardware for microscopy'.

Highly contractile 3D tissue engineered skeletal muscles from human iPSCs reveal similarities with primary myoblast-derived tissues.

Skeletal muscle research is transitioning toward 3D tissue engineered in vitro models reproducing muscle's native architecture and supporting measurement of functionality. Human induced pluripotent stem cells (hiPSCs) offer high yields of cells for differentiation. It has been difficult to differentiate high-quality, pure 3D muscle tissues from hiPSCs that show contractile properties comparable to primary myoblast-derived tissues. Here, we present a transgene-free method for the generation of purified, expandable myogenic progenitors (MPs) from hiPSCs grown under feeder-free conditions. We defined a protocol with optimal hydrogel and medium conditions that allowed production of highly contractile 3D tissue engineered skeletal muscles with forces similar to primary myoblast-derived tissues. Gene expression and proteomic analysis between hiPSC-derived and primary myoblast-derived 3D tissues revealed a similar expression profile of proteins involved in myogenic differentiation and sarcomere function. The protocol should be generally applicable for the study of personalized human skeletal muscle tissue in health and disease.

On the influence of protein aggregate sizes for the formation of solid and hollow protein microparticles.

Microparticles can function as carriers of e.g. pharmaceuticals and food ingredients. Hollow microparticles can enhance the capacitance due to their large interior void. For preparing microparticles, polymers have been assembled into spherical structures through the use of porous CaCO3 templates, followed by polymer cross-linking and selective template removal. However, this often results in the formation of microparticles with a solid core. Here we use proteins with different aggregate size distributions (<10 nm or >100 nm) to either form solid or hollow microparticles. Proteins were mixed with CaCl2 and Na2CO3 solutions, which from CaCO3 microcrystals (with 20-60 nm pores) with encapsulated proteins. Here it will be shown that small protein aggregates uniformly distributed into the CaCO3 templates. However, larger protein aggregates accumulated at the template edges. Au3+ ions were then added, which oxidize and cross-link proteins and are reduced to form gold nanoparticles (AuNPs). After removal of the templates, the small proteins formed solid microparticles and the larger protein aggregates hollow microparticles. This method of fabrication of solid and hollow protein microparticles, with embedded AuNPs, could be used for generating biomaterials with a broader range of applications, such as hosting molecules and multimodal imaging due to the presence of the AuNPs.

HistoEnder: A 3D printer-based histological slide autostainer that retains 3D printer functions.

Automated microscope slide stainers are usually very expensive and unless the laboratory performs heavy histological work it is difficult to justify buying a 2000-10000€ machine. As a result, histology and pathology labs around the world lose thousands of working hours for following procedures that could be easily automated. Herein, we propose a simple modification of an open-source 3D printer, the Creality Ender-3, into an automated microscope slide autostainer, the HistoEnder. The HistoEnder is cheap (less than 200€), modular, and easy to set up, with only two 3D-printed parts needed. Additionally, the 3D printer retains its full functionality, and it can be reverted back into 3D printing in less than 1 min. The g-code associated with the procedure is extremely simple, and can be written by anyone. The HistoEnder can also be used in chemistry and material science laboratories for automating surface modifications and dip coating.

A 3D Printer in the Lab: Not Only a Toy.

Although 3D printers are becoming more common in households, they are still under-represented in many laboratories worldwide and regarded as toys rather than as laboratory equipment. This short review wants to change this conservative point of view. This mini-review focuses on fused deposition modeling printers and what happens after acquiring your first 3D printer. In short, these printers melt plastic filament and deposit it layer by layer to create the final object. They are getting cheaper and easier to use, and nowadays it is not difficult to find good 3D printers for less than €500. At such a price, a 3D printer is one, if not the most, versatile piece of equipment you can have in a laboratory.

Hollow protein microparticles formed through cross-linking by an Au3+ initiated redox reaction.

Hollow microparticles (MPs) are of great relevance in the materials industry for a wide range of applications, such as catalysis, coatings, and delivery of theranostics. Here, we report the formation of hollow MPs through the assembly of lipoproteins in CaCO3 templates. Proteins interact in the pores of CaCO3 templates through attractive hydrophobic forces and form dense edges of hollow MPs. To further cross-link the proteins, Au3+ was added to initiate a redox reaction, where proteins were oxidized forming inter- and intramolecular covalent bonds, while Au3+ was reduced and gold nanoparticles (AuNPs) were formed. The obtained protein-based hollow MPs have a diameter of 6 µm and the AuNPs are embedded on their surface. Through this research, we suggest a new route to design biobased Au-protein hollow MPs in simple steps, which can allow new possibilities for carrying functional molecules and bioimaging.

T-Cup: A Cheap, Rapid, and Simple Home Device for Isothermal Nucleic Acid Amplification.

During the SARS-CoV2 pandemic, it has become clear that centralized testing suffers from multiple bottlenecks. Logistics, number of machines, and people available to run the diagnostic tests are limited. A solution to those bottlenecks would be a fully decentralized system, where people can test themselves at home and only report back the outcome of the test in a centralized database. Here a noninstrumental device capable of achieving isothermal conditions useful for detecting the SARS-CoV2 RNA using loop mediated amplification (LAMP) tests is presented. This device, compared to others reported in literature or present on the market, is cheap, easy to produce and use, and has little impact on the environment. Using a simple aluminum coffee capsule, a phase change material, and a 3D printed holder, this device, when placed in boiling water, is able to maintain a temperature of 65 °C for 25 min, required for running the LAMP reaction. In principle, this device can be applied to any LAMP reaction, and hence employed for many different applications, and can be deployed in large quantities in short amount of time.

The Field Guide to 3D Printing in Optical Microscopy for Life Sciences.

The maker movement has reached the optics labs, empowering researchers to create and modify microscope designs and imaging accessories. 3D printing has a disruptive impact on the field, improving accessibility to fabrication technologies in additive manufacturing. This approach is particularly useful for rapid, low-cost prototyping, allowing unprecedented levels of productivity and accessibility. From inexpensive microscopes for education such as the FlyPi to the highly complex robotic microscope OpenFlexure, 3D printing is paving the way for the democratization of technology, promoting collaborative environments between researchers, as 3D designs are easily shared. This holds the unique possibility of extending the open-access concept from knowledge to technology, allowing researchers everywhere to use and extend model structures. Here, it is presented a review of additive manufacturing applications in optical microscopy for life sciences, guiding the user through this new and exciting technology and providing a starting point to anyone willing to employ this versatile and powerful new tool.

Au3+-Induced gel network formation of proteins.

The formation of protein gel networks in aqueous systems is a result of protein intermolecular interactions after an energy input, like heating. In this research, we report that a redox reaction between Au3+ ions and proteins can also lead to the formation of a protein gel network. Amino acids, like cysteine and tyrosine, get oxidized and form covalent bonds with neighboring protein molecules, while Au3+ ions get reduced to Au+ and Au0, nucleate and form gold nanoparticles. The protein gel network formation occurs within 2 h at room temperature and can be tuned by varying Au3+/protein ratio and accelerated by increasing the incubation temperature. The proposed Au3+-induced gel network formation was applied to different proteins, like egg yolk high-density lipoprotein, bovine serum albumin and whey protein. This research opens new insights for the investigation of the metal-protein interactions and may aid in the design of novel hybrid-soft nanocomposite materials.

Experiments@home.

The coronavirus pandemic forced a rapid adoption of online learning. What can be done for teaching the practical elements of subjects like chemistry? Have we learned anything that we would keep outside of lockdown restrictions?

Hooked on mushrooms: Preparation and mechanics of a bioinspired soft probabilistic fastener.

Probabilistic fasteners are known to provide strong attachment onto their respective surfaces. Examples are Velcro® and the "3M dual lock" system. However, these systems typically only function using specific counter surfaces and are often destructive to other surfaces such as fabrics. Moreover, the design parameters to optimize their functionality are not obvious. Here, we present a surface patterned with soft micrometric features inspired by the mushroom shape showing a nondestructive mechanical interlocking and thus attachment to fabrics. We provide a scalable experimental approach to prepare these surfaces and quantify the attachment strength with rheometric and video-based analysis. In these "probabilistic fasteners," we find that higher feature densities result in higher attachment force; however, the individual feature strength is higher on a low feature density surface. We interpret our results via a load-sharing principle common in fiber bundle models. Our work provides new handles for tuning the mechanical attachment properties of soft patterned surfaces that can be used in various applications including soft robotics.

Ender3 3D printer kit transformed into open, programmable syringe pump set.

A cheap, open source 3D printer (Creality Ender 3) is transformed into an Open Hardware, programmable syringe pump set. Only 3 parts need to be purchased outside of the printer kit. All other parts are either in the Ender 3 kit, or can be 3D printed. No prior knowledge in electronics or programming languages is required. The pumps are controlled by the 3D printer firmware and motherboard and programmed in simple G-code text files. The total cost of a three pumps setup is ∼€170. The pumps are capable of reaching stable flows down to 5 µL/min using cheap, disposable 10 mL syringes. Higher flow speeds are also achievable, in the order of mL/min.

Response of metal-coordination-based polyelectrolyte complex micelles to added ligands and metals.

Polyelectrolyte complex based micelles have attracted significant attention due to their potential regarding bio-applications. Although the morphology and functions have been studied extensively, dynamic properties, particularly component exchange with other surrounding molecules, have remained elusive to date. Here, we show how micelles based on metal-ligand coordination complex coacervate-core micelles (M-C3Ms) respond to addition of extra ligand and metal ions. The micelles are prepared from a polycationic-neutral diblock copolymer and an anionic coordination polyelectrolyte, which is obtained by coordination between metal ions (lanthanides Ln3+ and Zn2+) and a bis-ligand (LEO) containing two dipicolinic acid (DPA) groups connected by a tetra-ethylene oxide spacer (4EO). Our findings show that the bis-ligand LEO is essential for the growth of coordination polymers and consequently the formation of micelles, leading to equilibrium structures with the same micellar composition and structure independent of the order of mixing. In other words, adding single DPA has no effect on the formed M-C3Ms. As for metal exchange, we find that added Zn2+ can replace some of the Ln3+ from Ln-C3Ms, leading to a hybrid coordination structure with both Ln3+ and Zn2+. We find that component exchange occurs in these coordination polyelectrolyte micelles, but it is more favorable in the direction of replacing the weak binding components with strong ones. Hence, the designed M-C3Ms based on the strong binding components, such as Ln-C3Ms, shall be relatively stable in biological surroundings, paving the way for the application of such particles as bio-imaging probes.

Gold and silver dichroic nanocomposite in the quest for 3D printing the Lycurgus cup.

The Lycurgus cup is an ancient glass artefact that shows dichroism as it looks green when a white light is reflected on it and a red colouring appears when a white light is transmitted through it. This peculiar dichroic effect is due to silver and gold nanoparticles present in the glass. In this research we show the synthesis of dichroic silver nanoparticles and their embedding in a 3D printable nanocomposite. The addition of gold nanoparticles to the silver nanoparticle composite, gave a 3D printable nanocomposite with the same dichroism effect of the Lycurgus cup.

COvalent monolayer patterns in Microfluidics by PLasma etching Open Technology - COMPLOT.

Plasma microcontact patterning (PµCP) and replica molding were combined to make PDMS/glass microfluidic devices with ß-cyclodextrin (ß-CD) patterns attached covalently on the glass surface inside microchannels. The supramolecular reactivity, reusability and association constant of ß-CD with Cy5-Ad2 was tested by analyzing signal-to-noise ratios of patterns vs. spacing with fluorescence microscopy.

Covalently bound monolayer patterns obtained by plasma etching on glass surfaces.

Micropatterns of ß-cyclodextrin (ß-CD) monolayers on glass are obtained by using a plasma etching approach with polydimethylsiloxane (PDMS) stamps. This simple and versatile approach provides a promising alternative to current techniques for creating patterns of covalently bound molecules. It is also possible to fabricate sub-10 µm sized features.

Gold nanoparticles embedded in a polymer as a 3D-printable dichroic nanocomposite material.

Background: Nanotechnology, even if unknowingly, has been used for millennia. The occurrence of shiny colors in pottery and glass made hundreds and thousand of years ago is due to the presence of nanoparticles in the fabrication of such ornaments. In the last decade, 3D printing has revolutionized fabrication and manufacturing processes, making it easier to produce, in a simple and fast way, 3D objects. Results: In this paper we show how to fabricate a 3D-printable nanocomposite composed of dichroic gold nanoparticles and a 3D-printable polymer. The minute amount of gold nanoparticles used for obtaining the dichroic effect does not influence the mechanical properties of the polymer nor its printability. Thus, the nanocomposite can be easily 3D-printed using a standard 3D printer and shows a purple color in transmission and a brownish color in reflection. Conclusion: This methodology can be used not only by artists, but also for studying the optical properties of nanoparticles or, for example, for the 3D fabrication of optical filters.

Cyclodextrin-based complex coacervate core micelles with tuneable supramolecular host-guest, metal-to-ligand and charge interactions.

Micelles have been recognized as versatile platforms for different biomedical applications, from bioimaging to drug delivery. Complex coacervate core micelles present great advantages compared to traditional micelles, however controlling the number of charges per core-unit and the stability is still a challenge. We here present cyclodextrin-based complex coacervate core micelles where the charge per core-unit can be straightforwardly tuned by cyclodextrin host-guest interactions. By varying the ratio between two adamantane guest molecules, 1-adamantanecarboxylic acid and 1,3-adamantanediacetic acid, the charge of the monomeric core-units can be finely tuned from 6- to 9-. By adding an adamantane bislinker, monomeric core-units can be combined together in dimeric and polymeric structures, increasing the micelles' stability. The orthogonal supramolecular host-guest and coordination-chemistry allows for well-controlled cyclodextrin-based complex coacervate core micelles that offer a versatile platform for designing future, e.g., responsive systems.

Loop-mediated isothermal amplification (LAMP) shield for Arduino DNA detection.

OBJECTIVE: Loop-mediated isothermal amplification (LAMP) of DNA is gaining relevance as a method to detect nucleic acids, as it is easier, faster, and more powerful than conventional Polymerase Chain Reaction. However, LAMP is still mostly used in laboratory settings, because of the lack of a cheap and easy, one-button device that can perform LAMP experiments. RESULTS: Here we show how to build and program an Arduino shield for a LAMP and detection of DNA. The here described Arduino Shield is cheap, easy to assemble, to program and use, it is battery operated and the detection of DNA is done by naked-eye so that it can be used in field.