Portal de Periódicos da CAPES

Strain Library Imaging Protocol for high-throughput, automated single-cell microscopy of large bacterial collections arrayed on multiwell plates

2017; Nature Portfolio; Volume: 12; Issue: 2 Linguagem: Inglês

10.1038/nprot.2016.181

1754-2189

Handuo Shi, Alexandre Colavin, Timothy K. Lee, Kerwyn Casey Huang,

Gene Regulatory Network Analysis

SLIP is a high-throughput, automated microscopy workflow for large strain collections. Bacterial cultures are transferred to large agar pads using replicator pins, and thousands of images are automatically acquired for single-cell quantification. Single-cell microscopy is a powerful tool for studying gene functions using strain libraries, but it suffers from throughput limitations. Here we describe the Strain Library Imaging Protocol (SLIP), which is a high-throughput, automated microscopy workflow for large strain collections that requires minimal user involvement. SLIP involves transferring arrayed bacterial cultures from multiwell plates onto large agar pads using inexpensive replicator pins and automatically imaging the resulting single cells. The acquired images are subsequently reviewed and analyzed by custom MATLAB scripts that segment single-cell contours and extract quantitative metrics. SLIP yields rich data sets on cell morphology and gene expression that illustrate the function of certain genes and the connections among strains in a library. For a library arrayed on 96-well plates, image acquisition can be completed within 4 min per plate.