Application of Imaging-Based Assays in Microplate Formats for High-Content Screening

  • Adam I. Fogel
  • Scott E. Martin
  • Samuel A. HassonEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1439)


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.

Key words

High-content screening High-content analysis Automated microscopy RNAi siRNA Functional genomics Imaging assay Assay development Image analysis 



We would like to thank Madhu Lal-Nag for assistance with image retrieval. This work was supported by the Intramural Research Program of the NIH, NINDS and the Trans-NIH RNAi initiative.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Adam I. Fogel
    • 2
  • Scott E. Martin
    • 3
  • Samuel A. Hasson
    • 1
    • 4
    Email author
  1. 1.National Center for Advancing Translational SciencesNational Institutes of HealthRockvilleUSA
  2. 2.BiogenCambridgeUSA
  3. 3.Department of Discovery Oncology Genentech Inc.South San FranciscoUSA
  4. 4.Pfizer, Inc.CambridgeUSA

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