A mercury arc lamp-based multi-color confocal real time imaging system for cellular structure and function

Cell Struct Funct. 2008;33(1):133-41. doi: 10.1247/csf.08015. Epub 2008 Aug 6.

Abstract

Multi-point scanning confocal microscopy using a Nipkow disk enables the acquisition of fluorescent images with high spatial and temporal resolutions. Like other single-point scanning confocal systems that use Galvano meter mirrors, a commercially available Nipkow spinning disk confocal unit, Yokogawa CSU10, requires lasers as the excitation light source. The choice of fluorescent dyes is strongly restricted, however, because only a limited number of laser lines can be introduced into a single confocal system. To overcome this problem, we developed an illumination system in which light from a mercury arc lamp is scrambled to make homogeneous light by passing it through a multi-mode optical fiber. This illumination system provides incoherent light with continuous wavelengths, enabling the observation of a wide range of fluorophores. Using this optical system, we demonstrate both the high-speed imaging (up to 100 Hz) of intracellular Ca(2+) propagation, and the multi-color imaging of Ca(2+) and PKC-gamma dynamics in living cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium / metabolism
  • Cell Nucleus / metabolism
  • Cellular Structures / physiology
  • Cellular Structures / ultrastructure*
  • Computer Systems
  • Cytoplasm / metabolism
  • Fiber Optic Technology
  • Fluorescent Dyes
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Lasers
  • Light
  • Luminescent Proteins / analysis
  • Mercury / chemistry*
  • Microscopy, Confocal / instrumentation
  • Microscopy, Confocal / methods*
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Mitochondria / metabolism
  • Optical Fibers

Substances

  • Fluorescent Dyes
  • Luminescent Proteins
  • Mercury
  • Calcium