Development and implementation of multiplexed cell-based imaging assays.
Fluorescence microscopy, image analysis, and automated screening technologies are some of the most powerful tools enabling cell biologists to investigate complex signaling pathways and compound or siRNA effects on cellular function in individual cells. Researchers can now use multiple fluorescent probes to quantify effects on intracellular molecular events, measure phenotypic changes, and provide contextual information about cellular pathways not discernible by traditional single-parameter, end point experiments. This chapter focuses on fluorescent labeling techniques and methods for designing image-based assays, multiplexed readouts, and image analysis routines. Case studies are presented describing the use of cell-based imaging assays for monitoring cell proliferation, cell cycle stage, and apoptosis.
Howell
Bonnie J
BJ
Department of Cancer Research, Merck Research Laboratories, West Point, PA, USA.
Lee
Seungtaek
S
Sepp-Lorenzino
Laura
L
D000818:Animals; D017209:Apoptosis; D001681:Biological Assay / Q000379:methods*; D002453:Cell Cycle; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005456:Fluorescent Dyes / Q000494:pharmacology; D006657:Histones / Q000737:chemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D020287:In Situ Nick-End Labeling; D008856:Microscopy, Fluorescence / Q000379:methods*; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction
D005456:Fluorescent Dyes; D006657:Histones; D034741:RNA, Small Interfering