"Microscopy and Image Analysis"
Microscopy and Image Analysis UNIT 4.4 George McNamara,1 Michael Difilippantonio,2 Thomas Ried,3 and Frederick R. Bieber4 1Biomedical Consultant, Baltimore, Maryland 2Division of Cancer Treatment and Diagnosis, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 3Section of Cancer Genomics, Genetics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 4Brigham and Women’s Hospital, Boston, Massachusetts This unit provides an overview of light microscopy, including objectives, light sources, filters, film, and color photography for fluorescence microscopy and fluorescence in situ hybridization (FISH). We believe there are excellent oppor- tunities for cytogeneticists, pathologists, and other biomedical readers, to take advantage of specimen optical clearing techniques and expansion microscopy— we briefly point to these new opportunities. C 2017 by John Wiley & Sons, Inc. Keywords: light microscopy r digital imaging r fluorescence in situ hybridiza- tion r functional genomics How to cite this article: McNamara, G., Difilippantonio, M., Ried, T., & Bieber, F. R. (2017). Microscopy and image analysis. Current Protocols in Human Genetics, 94, 4.4.1–4.4.89. doi: 10.1002/cphg.42 INTRODUCTION by Ishikawa-Ankerhold, Ankerhold, & Drum- This unit provides an overview of light mi- men (2012), and Liu, Ahmed, & Wohland croscopy, including objectives, light sources, (2008). Scanning and transmission electron filters, and imaging for fluorescence mi- microscopy as well as confocal microscopy croscopy and fluorescence in situ hybridiza- and multi-photon excitation microscopy are tion (FISH). We encourage thinking outside not covered in this unit despite their useful- the usual magnification range of 10× to ness as invaluable tools for contemporary stud- 100× objective lenses, by ranging from sin- ies of biological systems; see Diaspro, 2001; gle molecules to whole mice and humans.
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