Basics of Digital Microscopy

Callen T. Wallace1, Morgan Jessup1, Tytus Bernas2, Karina A. Peña1, Michael J. Calderon1, Patricia A. Loughran1

1 University of Pittsburgh—Center for Biologic Imaging, Pittsburgh, Pennsylvania, 2 Purdue University, West Lafayette, Indiana
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 12.2
DOI:  10.1002/cpcy.31
Online Posting Date:  January, 2018
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Abstract

Modern digital microscopy combines the equipment of classical light microscopy with a computerized imaging system. The technique comprises image formation by optics, image registration by a camera, and saving of image data in a computer file. This chapter describes limitations that are particular to each of these processes, including optical resolution, efficiency of image registration, characteristics of image file formats, and data management. Further suggestions are given which serve, in turn, to help construct a set of guidelines aimed at optimization of digital microscopic imaging. © 2018 by John Wiley & Sons, Inc.

Keywords: image file format; quantization; resolution; sampling; signal‐to‐noise ratio; compression

     
 
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Table of Contents

  • Optical Resolution
  • Digital Registration of an Optical Image
  • Storage of Digital Image Data
  • Conclusions
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Figures

Videos

Literature Cited

Literature Cited
  Bernas, T. (2005). Basics of digital microscopy. Current Protocols in Cytometry, 31, 12.2:12.2.1–12.2.14. doi: 10.1002/0471142956.cy1202s31.
  Gonzalez, R. C., & Woods, R. F. (2002). Digital image processing (2nd ed.). Upper Saddle River, NJ.: Prentice Hall.
  Houston, W. V. (1926). The fine structure and wave‐length of the Balmer Lines. The Astrophysical Journal, 64, 81. doi: 10.1086/142989.
  Joubert, J., & Sharma, D. (2011). Light microscopy digital imaging. Current Protocols in Cytometry, 58, 2.3:2.3.1–2.3.11. doi: 10.1002/0471142956.cy0203s58.
  Rayleigh, J. W. S. (1879). Investigations in Optics with special reference to the Spectroscope. Philosophical Magazine, 8, 261–274. doi: 10.1080/14786447908639684.
  Sparrow, C. M. (1916). On spectroscopic resolving power. The Astrophysical Journal, 44, 76. doi: 10.1086/142271.
  Sysko, L. R., & Davis, M. A. (2010). From image to data using common image‐processing techniques. Current Protocols in Cytometry, 54, 12.21:12.21.1–12.21.17. doi: 10.1002/0471142956.cy1221s54
  Young, I. T. (2001). Calibration: Sampling density and spatial resolution. Current Protocols in Cytometry, 5, 2.6:2.6.1–2.6.15. doi: 10.1002/0471142956.cy0206s05.
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