Abstract
.Fiber bundle fluorescence endomicroscopy is an effective method for in vivo imaging of biological tissue samples. Line-scanning confocal laser endomicroscopy realizes confocal imaging at a much higher frame rate compared to the point scanning system, but with reduced optical sectioning. To address this problem, we describe a fiber bundle endomicroscopy system that utilizes the HiLo technique to enhance the optical sectioning while still maintaining high image acquisition rates. Confocal HiLo endomicroscopy is achieved by synchronizing the scanning hybrid-illumination laser line with the rolling shutter of a CMOS camera. An evident improvement of axial sectioning is achieved as compared to the line-scanning confocal endomicroscopy without the HiLo technique. Comparisons are also made with epifluorescence endomicroscopy with and without HiLo. The optical sectioning enhancement is demonstrated on lens tissue as well as porcine kidney tissue.
Highlights
Optical fiber bundles represent an important element for implementing endomicroscopy for flexible maneuvering along a curved lumen and providing microscopic imaging of a living tissue.[1,2,3] Fiber bundles have been used in various single-photon imaging modalities, such as epifluorescence endomicroscopy and confocal fluorescence endomicroscopy.[1,2,3,4,5] The epifluorescence endomicroscopy acquires signal from all pixels in parallel to provide the highest acquisition speeds
We present an implementation of a high-speed fiber bundle fluorescence endomicroscope that demonstrates enhanced optical sectioning by combining the benefits of HiLo technique and confocal slit detection
The results demonstrated that the optical sectioning ability of the HiLo-confocal endomicroscopy is largely improved when compared to other configurations
Summary
Optical fiber bundles represent an important element for implementing endomicroscopy for flexible maneuvering along a curved lumen and providing microscopic imaging of a living tissue.[1,2,3] Fiber bundles have been used in various single-photon imaging modalities, such as epifluorescence endomicroscopy and confocal fluorescence endomicroscopy.[1,2,3,4,5] The epifluorescence endomicroscopy acquires signal from all pixels in parallel to provide the highest acquisition speeds. Confocal fluorescence endomicroscopy provides an effective means of eliminating signals from out-of-focus light by focusing laser illumination to a single point and the use of a small pinhole in an optical plane conjugate.[3,6] It allows optical sectioning of cells and tissue and has found a wide range of clinical applications, including in vivo imaging of the urinary tract as well as the human airway,[7,8] and diagnosis of colorectal cancer.[9]
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