Biology
Latest
Optogenetic control of Nodal signaling patterns
Embryos issue instructions to their cells in the form of patterns of signaling activity. Within these patterns, the distribution of signaling in time and space directs the fate of embryonic cells. Tools to perturb developmental signaling with high resolution in space and time can help reveal how these patterns are decoded to make appropriate fate decisions. In this talk, I will present new optogenetic reagents and an experimental pipeline for creating designer Nodal signaling patterns in live zebrafish embryos. Our improved optoNodal reagents eliminate dark activity and improve response kinetics, without sacrificing dynamic range. We adapted an ultra-widefield microscopy platform for parallel light patterning in up to 36 embryos and demonstrated precise spatial control over Nodal signaling activity and downstream gene expression. Using this system, we demonstrate that patterned Nodal activation can initiate specification and internalization movements of endodermal precursors. Further, we used patterned illumination to generate synthetic signaling patterns in Nodal signaling mutants, rescuing several characteristic developmental defects. This study establishes an experimental toolkit for systematic exploration of Nodal signaling patterns in live embryos.
PiSpy: An Affordable, Accessible, and Flexible Imaging Platform for the Automated Observation of Organismal Biology and Behavior
A great deal of understanding can be gleaned from direct observation of organismal growth, development, and behavior. However, direct observation can be time consuming and influence the organism through unintentional stimuli. Additionally, video capturing equipment can often be prohibitively expensive, difficult to modify to one’s specific needs, and may come with unnecessary features. Here, we describe the PiSpy, a low-cost, automated video acquisition platform that uses a Raspberry Pi computer and camera to record video or images at specified time intervals or when externally triggered. All settings and controls, such as programmable light cycling, are accessible to users with no programming experience through an easy-to-use graphical user interface. Importantly, the entire PiSpy system can be assembled for less than $100 using laser-cut and 3D-printed components. We demonstrate the broad applications and flexibility of the PiSpy across a range of model and non-model organisms. Designs, instructions, and code can be accessed through an online repository, where a global community of PiSpy users can also contribute their own unique customizations and help grow the community of open-source research solutions.
Addgene AAV data hub
The Addgene AAV Data Hub was launched to help scientists share data and protocols obtained from AAV experiments. Our longterm goal is to provide scientists with a resource to help guide AAV selection and use by providing data from individual labs on AAV performance.
An open-source experimental framework for automation of cell biology experiments
Modern biological methods often require a large number of experiments to be conducted. For example, dissecting molecular pathways involved in a variety of biological processes in neurons and non-excitable cells requires high-throughput compound library or RNAi screens. Another example requiring large datasets - modern data analysis methods such as deep learning. These have been successfully applied to a number of biological and medical questions. In this talk we will describe an open-source platform allowing such experiments to be automated. The platform consists of an XY stage, perfusion system and an epifluorescent microscope with autofocusing. It is extremely easy to build and can be used for different experimental paradigms, ranging from immunolabeling and routine characterisation of large numbers of cell lines to high-throughput imaging of fluorescent reporters.
Biology coverage
4 items