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A famous example from the late David Rogers at Vanderbilt University in the 1950s shows a human neutrophil in a blood smear chasing a Staphylococcus aureus bacterium and eventually the process of phagocytosis 4. From the earliest microscopes, enthusiasts and scientists have observed and studied single cells and organisms, their behaviors, and development 1-3. Live-cell microscopy and longitudinal tracking of single cells is not a new technique.

We also discuss variations of the technique and its flexibility, and highlight some common pitfalls. We show examples that include tracking and quantification of cell growth and cell cycle progression, chromosome dynamics, DNA damage, and cell death. With the employment of genetically encoded fluorescent biosensors numerous processes, pathways and responses can be followed. Using variations of the approach, cells can be monitored for weeks. We describe an approach in detail that allows for the continuous time-lapse imaging of cells during the drug response for essentially as long as desired, typically up to 96 hr. Advancements in technology enable us to routinely image cells at a resolution that enables not only cell tracking, but also the observation of a variety of cellular responses. The microscope is particularly well suited to image live cells. Variability in drug responses between cancer and normal cells, and between cells of different cancer origin, and transient and rare responses are difficult to understand using population averaging assays and without being able to directly track and analyze them longitudinally. These methods are important, but they have several shortcomings. Immunoblotting, flow cytometry and fixed cell experiments are often used to study how cells respond to anti-cancer drugs. The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing factor in the overall outcome.

How to slow down adobe illustrator draws time lapse video