Basics of Digital Microscopy

Tytus Bernas1

1 Purdue University, West Lafayette, Indiana
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 12.2
DOI:  10.1002/0471142956.cy1202s31
Online Posting Date:  February, 2005
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Abstract

Digital microscopy combines the equipment of classic light microscopy with a computerized imaging system, i.e., a digital camera. This 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, and characteristics of image file formats. Knowledge with regard to these limitations serves, in turn, to help construct a set of guidelines aimed at optimization of digital microscopic imaging.

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

     
 
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Table of Contents

  • Optical Imaging
  • 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
   Gonzalez, R.C. and Woods, R.F. 2002. Digital Image Processing, 2nd ed. Prentice Hall, Upper Saddle River, N.J.
   Holmes, T.J. 1992. Blind deconvolution of quantum‐limited incoherent imagery: Maximum‐likelihood approach. J. Opt. Soc. Amer. A‐9:1052‐1061.
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   Holmes, T.J. and Liu, Y.‐H. 1991. Acceleration of maximum‐likelihood image restoration for fluorescence microscopy and other incoherent imagery. J. Opt. Soc. Amer. A‐8:893‐907.
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   Reichman, J. 2000. Handbook of Optical Filters for Fluorescence Microscopy. Chroma Technology Corp., Rockingham, Vt.
   Sparrow, C. M. 1916. On spectroscopic resolving power. Astrophys. J. 44:76.
   Stelzer, E.H.K. 1997. Contrast, resolution, pixelation, dynamic range and signal‐to‐noise ratio: Fundamental limits of resolution in fluorescence light microscopy. J. Microsc. 189:15‐24.
   Van Dekker, A.J. and Bos, A. 1997. Resolution: A survey. J. Opt. Soc. Amer. A‐14:547‐557.
   Young, I.T. 1996. Quantitative microscopy. IEEE Eng. Med. Biol. 15:59‐66.
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