Confocal fluorescence microscope with dual-axis architecture and biaxial postobjective scanning.
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.
Wang
Thomas D
TD
Stanford University School of Medicine, Stanford, California 94305, USA. tdwang@stanford.edu
Contag
Christopher H
CH
Mandella
Michael J
MJ
Chan
Ning Y
NY
Kino
Gordon S
GS
D000818:Animals; D002531:Cerebellum / Q000166:cytology; D004330:Drosophila / Q000166:cytology* / Q000196:embryology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D007089:Image Enhancement / Q000295:instrumentation*; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D018482:Muscle, Skeletal / Q000166:cytology*; D009474:Neurons / Q000166:cytology*