A line scanning confocal fluorescent microscope using a CMOS rolling shutter as an adjustable aperture

J Microsc. 2012 Sep;247(3):269-76. doi: 10.1111/j.1365-2818.2012.03642.x.

Abstract

Traditional confocal microscopy uses a physical aperture barrier to prevent out-of-focus light from reaching the detector. The physical nature of a conventional aperture limits control over the system confocality. We describe a new line scanning confocal microscope that eliminates a need for a physical aperture by employing a software-controllable rolling shutter on a CMOS camera. A confocal image is obtained by synchronizing motion of the rolling shutter and the laser line scanning over a sample. Confocal resolution of this microscope is adjustable in real time and independently established for each fluorescence channel by changing the rolling shutter width. This technology has been implemented in the IN Cell Analyzer 6000 system by GE Healthcare.

MeSH terms

  • Animals
  • CHO Cells
  • Cell Nucleus / chemistry
  • Cricetinae
  • Fluorescence
  • Image Processing, Computer-Assisted / instrumentation*
  • Image Processing, Computer-Assisted / methods
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods
  • Microscopy, Confocal / instrumentation*
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Reproducibility of Results
  • Sensitivity and Specificity
  • User-Computer Interface*