Microscopy and Image Analysis

George McNamara1, Michael Difilippantonio2, Thomas Ried2, Frederick R. Bieber3

1 Biomedical Consultant, Baltimore, Maryland, 2 National Cancer Institute, National Institutes of Health, Bethesda, 3 Brigham and Women's Hospital, Boston, Massachusetts
Publication Name:  Current Protocols in Human Genetics
Unit Number:  Unit 4.4
DOI:  10.1002/cphg.42
Online Posting Date:  July, 2017
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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 opportunities 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. © 2017 by John Wiley & Sons, Inc.

Keywords: light microscopy; digital imaging; fluorescence in situ hybridization; functional genomics

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Table of Contents

  • Introduction
  • Nano, Micro, And Macro Scales
  • Historical Foundations of Microscopy
  • Microscopy in Modern Human Genetics
  • Types of Microscopy
  • Microscope Objectives and Eyepiece Lenses
  • Fluorescence Microscopy
  • Light Sources
  • From Fluorescent Dyes, Qdots, and Polymers to Fluorescent Proteins
  • Image Acquisition
  • Conclusions
  • Supporting Materials
  • Acknowledgments
  • Conflict of Interest Statement
  • Figures
  • Tables
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Key References
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  See supplemental file. An extensive review related to the construction and use of fluorescently tagged proteins for monitoring and measuring proteins and protein‐protein interactions in living cells.
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  Fluorescent biosensor applications for live cell imaging of many biological processes.
Internet Resources
  Contributed by DPB/DGE Codex; see W. B. Frommer entry for molecular sensors.
  Contributed by Addgene; fluorescent protein guide: Biosensors.
  Contributed by George McNamara.
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