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 scanni...

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Bibliographic Details
Published in:Journal of Biomedical Optics 2004-07, Vol.9 (4), p.735-742
Main Authors: Wang, Thomas D, Contag, Christopher H, Mandella, Michael J, Chan, Ning Y, Kino, Gordon S
Format: Article
Language:English
Subjects:
Animals
Cerebellum - cytology
confocal
Drosophila - cytology
Drosophila - embryology
Equipment Design
Equipment Failure Analysis
Feasibility Studies
fluorescence
Image Enhancement - instrumentation
imaging
Mice
microscope
Microscopy, Confocal - instrumentation
Microscopy, Confocal - methods
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
molecular biology
Muscle, Skeletal - cytology
Neurons - cytology
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Description
Summary: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 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 molecular and cellular imaging. ©
ISSN:1083-3668
1560-2281
DOI:10.1117/1.1760760