Application of Imaging-Based Assays in Microplate Formats for High-Content Screening.
The use of multiparametric microscopy-based screens with automated analysis has enabled the large-scale study of biological phenomena that are currently not measurable by any other method. Collectively referred to as high-content screening (HCS), or high-content analysis (HCA), these methods rely on an expanding array of imaging hardware and software automation. Coupled with an ever-growing amount of diverse chemical matter and functional genomic tools, HCS has helped open the door to a new frontier of understanding cell biology through phenotype-driven screening. With the ability to interrogate biology on a cell-by-cell basis in highly parallel microplate-based platforms, the utility of HCS continues to grow as advancements are made in acquisition speed, model system complexity, data management, and analysis systems. This chapter uses an example of screening for genetic factors regulating mitochondrial quality control to exemplify the practical considerations in developing and executing high-content campaigns.
Biogen, Cambridge, MA, 02142, USA.
Adam I
AI
Fogel
Department of Discovery Oncology Genentech Inc., South San Francisco, CA, 94080, USA.
Scott E
SE
Martin
National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA. Samuel.Hasson@pfizer.com.|Pfizer, Inc., 610 Main Street, Cambridge, MA, 02144, USA. Samuel.Hasson@pfizer.com.
Samuel A
SA
Hasson
D000818:Animals; D023281:Genomics / Q000379:methods*; D057809:HEK293 Cells; D006367:HeLa Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008928:Mitochondria / Q000235:genetics; D061848:Optical Imaging / Q000379:methods*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012984:Software
D034741:RNA, Small Interfering
Assay development; Automated microscopy; Functional genomics; High-content analysis; High-content screening; Image analysis; Imaging assay; RNAi; siRNA