Live-cell Imaging and Analysis of Germline Stem Cell Mitosis in .

Model organisms offer the opportunity to decipher the dynamic and complex behavior of stem cells in their native environment; however, imaging stem cells remains technically challenging. germline stem cells (GSCs) are distinctly accessible for live imaging but relatively few studies have taken advantage of this potential. Here we provide our protocol for mounting and live imaging dividing GSCs, as well as analysis tools to facilitate the processing of large datasets.

CentTracker: a trainable, machine-learning-based tool for large-scale analyses of germline stem cell mitosis.

Investigating the complex interactions between stem cells and their native environment requires an efficient means to image them in situ. germline stem cells (GSCs) are distinctly accessible for intravital imaging; however, long-term image acquisition and analysis of dividing GSCs can be technically challenging. Here we present a systematic investigation into the technical factors impacting GSC physiology during live imaging and provide an optimized method for monitoring GSC mitosis under minimally disruptive conditions.