Confocal fluorescence microscope with dual-axis architecture and biaxial postobjective scanning

J Biomed Opt. 2004 Jul-Aug;9(4):735-42. doi: 10.1117/1.1760760.

Abstract

We present a novel confocal microscope that has dual-axis architecture and biaxial postobjective scanning for the collection of fluorescence images from biological specimens. This design uses two low-numerical-aperture lenses to achieve high axial resolution and long working distance, and the scanning mirror located distal to the lenses rotates along the orthogonal axes to produce arc-surface images over a large field of view (FOV). With fiber optic coupling, this microscope can potentially be scaled down to millimeter dimensions via microelectromechanical systems (MEMS) technology. We demonstrate a benchtop prototype with a spatial resolution < or =4.4 microm that collects fluorescence images with a high SNR and a good contrast ratio from specimens expressing GFP. Furthermore, the scanning mechanism produces only small differences in aberrations over the image FOV. These results demonstrate proof of concept of the dual-axis confocal architecture for in vivo molecular and cellular imaging.

Publication types

  • Evaluation Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cerebellum / cytology
  • Drosophila / cytology*
  • Drosophila / embryology*
  • Equipment Design
  • Equipment Failure Analysis
  • Feasibility Studies
  • Image Enhancement / instrumentation*
  • Mice
  • Microscopy, Confocal / instrumentation*
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / instrumentation*
  • Microscopy, Fluorescence / methods
  • Muscle, Skeletal / cytology*
  • Neurons / cytology*