Entering the Portal: Understanding the Digital Image Recorded Through a Microscope

  • Kristin L. Hazelwood
  • Scott G. Olenych
  • John D. Griffin
  • Judith A. Cathcart
  • Michael W. Davidson
Chapter
Part of the Principles and Practice book series (PRINCIPLES)

The primary considerations in imaging living cells in the microscope with a digital camera are detector sensitivity (signal-to-noise), the required speed of image acquisition, and specimen viability. The relatively high light intensities and long exposure times that are typically employed in recording images of fixed cells and tissues (where photobleaching is the major consideration) must be strictly avoided when working with living cells. In virtually all cases, live-cell microscopy represents a compromise between achieving the best possible image quality and preserving the health of the cells. Rather than unnecessarily oversampling time points and exposing the cells to excessive levels of illumination, the spatial and temporal resolutions set by the experiment should be limited to match the goals of the investigation. This chapter describes the fundamentals of digital image acquisition, spatial resolution, contrast, brightness, bit depth, dynamic range, and CCD architecture, as well as performance measures, image display and storage, and imaging modes in optical microscopy.

Keywords

Fluorescence Resonance Energy Transfer Modulation Transfer Function Differential Interference Contrast High Spatial Frequency Fluorescence Recovery After Photobleaching 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer-Verlag Berlin Heidelberg 2007

Authors and Affiliations

  • Kristin L. Hazelwood
    • 1
  • Scott G. Olenych
    • 1
  • John D. Griffin
    • 1
  • Judith A. Cathcart
    • 1
  • Michael W. Davidson
    • 1
  1. 1.Optical Microscopy, National High Magnetic Field LaboratoryThe Florida State UniversityTallahasseeUSA

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