High-Throughput Imaging of Bacillus subtilis.

Bacillus subtilis is a widely used model bacterium to study cellular processes and development. The availability of an arrayed mutant library gave us the opportunity to cytologically analyze every mutant and screen for new genes involved in cell shape determination, cell division, and chromosome segregation. Here we describe a high-throughput method to image arrayed B. subtilis mutant libraries using wide-field fluorescence microscopy. We provide a detailed description of growing the arrayed strain collection, preparing slides containing agarose pedestals, setting up the microscopy procedure, acquiring images, and analyzing the images.

Best practices and tools for reporting reproducible fluorescence microscopy methods.

Although fluorescence microscopy is ubiquitous in biomedical research, microscopy methods reporting is inconsistent and perhaps undervalued. We emphasize the importance of appropriate microscopy methods reporting and seek to educate researchers about how microscopy metadata impact data interpretation. We provide comprehensive guidelines and resources to enable accurate reporting for the most common fluorescence light microscopy modalities. We aim to improve microscopy reporting, thus improving the quality, rigor and reproducibility of image-based science.