title: Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics.
abstract: 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. Immunoblotting, flow cytometry and fixed cell experiments are often used to study how cells respond to anti-cancer drugs. These methods are important, but they have several shortcomings. 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. The microscope is particularly well suited to image live cells. 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. 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. Using variations of the approach, cells can be monitored for weeks. With the employment of genetically encoded fluorescent biosensors numerous processes, pathways and responses can be followed. We show examples that include tracking and quantification of cell growth and cell cycle progression, chromosome dynamics, DNA damage, and cell death. We also discuss variations of the technique and its flexibility, and highlight some common pitfalls.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder.
authors.forename: Russell T
authors.initials: RT
authors.lastname: Burke
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder; James.Orth@colorado.edu.
authors.forename: James D
authors.initials: JD
authors.lastname: Orth
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D058948:Cell Tracking; D006801:Humans; D008853:Microscopy; D059010:Single-Cell Analysis; D059008:Time-Lapse Imaging
chemical_list: D000970:Antineoplastic Agents