Automated Cell Tracking
Our custom tracking software is capable of tracking thousands of cells simultaneously for multiple days. We primarily use fluorescently tagged Histone 2B to track and segment cells. This also allows us to automatically call mitosis when the condensed chromosomes divide into two daughter nuclei.
Cells can be tracked for multiple generations, allowing us to see how cells from a common ancestor behave similarly or differently.
Here we are tracking hundreds of HeLa cells for more than 40 hours. The cells trajectory is super imposed on the image, and every time a mitosis is detected, a red dot appears.
Automated Image Analysis
We image cells containing between 1 and 4 fluorescent biosensors in 96 well plates. Custom Matlab scripts are used to segment each cell and track their movements between frames. Multiple parameters are recorded for each cell and at each timepoint, including cell size, nuclear content, its proximity to neighbors, cell speed, as well as a quantification of each biosensor.
A ring around the nucleus is used to quantify cytoplasmic fluorescence levels.
Nuclear proteins can be easily quantified using a mask generated from H2B-CFP
Automated segmentation and spot identification
Before: Single-molecule mRNA FISH staining of cells expressing a fluorescent nuclear marker
After: Automated segmentation and spot identification using custom designed software
Live-cell imaging can be combined with fixed-cell analysis. This allows us to generate "time course" data for any method that involves cell fixation, such as immunofluorescence and FISH.