Andrew Pelling | bioRxiv Science

bioRxiv2016

Utilizing Social Media and Video Games to Control #DIY Microscopes

Open-source lab equipment is becoming more widespread with the popularization of fabrication tools such as 3d-printers, laser cutters, CNC machines, open source microcontrollers and open source software. Although many pieces of common laboratory equipment have been developed, software control of these items is sometimes lacking. Specifically, control software that can be easily implemented and enable user-input and control over multiple platforms (PC, smartphone, web, etc.). The aim of this proof-of-principle study was to develop and implement software for the control of a low-cost, 3d-printed microscope. Here, we present two approaches, which enable microscope control by exploiting the functionality of the social media platform Twitter or player actions inside of the videogame Minecraft. The microscope was constructed from a modified web-camera and implemented on a Raspberry Pi computer. Four aspects of microscope control were tested, including single image capture, focus control and time-lapse imaging. The Twitter-embodiment enabled users to send \"tweets\" directly to the microscope. Image data acquired by the microscope was then returned to the user through a Twitter reply and stored permanently on the photo-sharing platform Flickr, along with any relevant metadata. Local control of the microscope was also implemented by utilizing the video game Minecraft, in situations where Internet connectivity is not present or stable. A virtual laboratory was constructed inside the Minecraft world and player actions inside the laboratory were linked to specific microscope functions. Here, we present the methodology and results of these experiments and discuss possible limitations and future extensions of this work.

Biophysics↗

bioRxiv2016

Measuring Mechanodynamics using an Unsupported Epithelial Monolayer Grown at an Air-Water Interface

Actomyosin contraction and relaxation in a monolayer is a fundamental biophysical process in development and homeostasis. Current methods used to characterize the mechanodynamics of monolayers often involve cells grown on solid supports such as glass or gels. The results of these studies are fundamentally influenced by these supporting structures. Here, we describe a new methodology for measuring the mechanodynamics of epithelial monolayers by culturing cells at an air-liquid interface. These model monolayers are grown in the absence of any supporting structures allowing us to remove cell-substrate effects. This methods potential was evaluated by observing and quantifying the generation and release of internal stresses upon actomyosin contraction (320{+/-}50Pa) and relaxation (190{+/-}40Pa) in response to chemical treatments. This is in contrast to the results observed in monolayers grown on solid substrates (glass and gels) where movement was drastically muted. Tracking the displacement of cell nuclei, cell edges and cluster perimeter allowed us to quantify the strain dynamics in the monolayer indicating the reliability of this method. New insights were also revealed with this approach. Although unsupported monolayers exhibited clear major and minor strain axes, they were not correlated to the general alignment of cell nuclei. The situation was dramatically different when the monolayers were grown on soft gels and hard glass substrates. It was observed that both gels and glass substrates led to the promotion of long-range alignment of cell nuclei. In addition, the strain orientation was correlated to nuclear alignment on the soft deformable gels. This new approach provides us with a picture of basal actomyosin mechanodynamics in a simplified system allowing us to infer how the presence of a substrate impacts actomyosin contractility and long-range multi-cellular organization and dynamics. This new methodology will also enable many new questions to be asked about the molecular regulation of the mechanodynamics of unsupported monolayers.

Biophysics↗