Spinning-disk confocal microscopy: present technology and future trends

Methods Cell Biol. 2014:123:153-75. doi: 10.1016/B978-0-12-420138-5.00009-4.

Abstract

Live-cell imaging requires not only high temporal resolution but also illumination powers low enough to minimize photodamage. Traditional single-point laser scanning confocal microscopy (LSCM) is generally limited by both the relatively slow speed at which it can acquire optical sections by serial raster scanning (a few Hz) and the higher potential for phototoxicity. These limitations have driven the development of rapid, parallel forms of confocal microscopy, the most popular of which is the spinning-disk confocal microscope (SDCM). Here, we briefly introduce the SDCM technique, discuss its strengths and weaknesses against LSCM, and update the reader on some recent developments in SDCM technology that improve its performance and expand its utility for life science research now and in the future.

Keywords: Borealis; Confocal microscopy; EMCCD camera; Fluorescence saturation; High-speed imaging; Multimode fiber; Phototoxicity; Single-mode fiber; Spinning disk; sCMOS camera.

Publication types

  • Review

MeSH terms

  • Animals
  • Humans
  • Image Processing, Computer-Assisted / instrumentation*
  • Image Processing, Computer-Assisted / standards
  • Image Processing, Computer-Assisted / trends
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / standards
  • Microscopy, Confocal / trends
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / standards
  • Microscopy, Fluorescence / trends
  • Quality Improvement
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / standards
  • Single-Cell Analysis / trends