By Haney, Steven A
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Current opinion in Microbiology, 2003, 6(3): 302–309. 16. NCCLS, In Situ Hybridization (FISH) Methods for Medical Genetics; Approved Guideline. (NCCLS documentMM7-A (ISBN 1-56238-524-0)). 17. , et al. Phototoxicity of the fluorescent membrane dyes PKH2 and PKH26 on the human hematopoietic KG1a progenitor cell line. Cytometry, 1999, 36(4): 312–318. 18. Pietraszewska-Bogiel, A. W. Gadella, FRET microscopy: From principle to routine technology in cell biology, 2011. 19. , Fluorescence lifetime imaging–techniques and applications.
In such a case, a scientist should not be dissuaded from proposing a high content experiment, but needs to collaborate with a microscopist. The roles of HCS instrumentation specialist, image and data analyst can be combined in drug screening groups. A high throughput screening (HTS) group in a pharmaceutical company typically manages many instruments, and works with biologists to adapt assays for HTS, but this can come at the expense of flexibility, as rapid progression through many screens can limit the time that can be spent on cell lines or imaging challenges for a particular project.
The breadth of options in labeling cells carries significant implications for the options in analyzing the data generated in an HCS assay. This includes integrating several events as a single assay through multiplexing, and capturing multiple measurements of the cellular architecture in a phenotypic profiling study. These topics will be covered in depth in later chapters, but the foundation for all of these studies lie in the ability to identify the components of a cell specifically and robustly.
Introduction To High Content Screening by Haney, Steven A