Photophysics of fluorescence, and applications of specialized F-techniques, are well reviewed by Ishikawa-Ankerhold, Ankerhold, & Drummen ( 2012), and Liu, Ahmed, & Wohland ( 2008). We discuss how functional genomics can contribute to cytogenetics and vice versa. Computerized image analysis systems currently used in clinical cytogenetics are also discussed, and connected with the larger trend of digital slide scanning in pathology and cytology. Genome engineering with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9, transcription activation-like effector nucleases (TALENs), and zinc finger nucleases requires both on-target validation and off-target safety checks.
Single molecule RNA FISH is now a research tool, and has opportunities in the clinic. Traditionally, clinical FISH was chromosome and single or dual gene DNA FISH. We encourage thinking outside the usual magnification range of 10× to 100× objective lenses, by ranging from single molecules to whole mice and humans.
This unit provides an overview of light microscopy, including objectives, light sources, filters, and imaging for fluorescence microscopy and fluorescence in situ hybridization (FISH).
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