Abstract
Confocal fluorescence microendoscopy provides high-resolution cellular-level imaging via a minimally invasive procedure, but requires fast scanning to achieve real-time imaging in vivo. Ideal confocal imaging performance is obtained with a point scanning system, but the scan rates required for in vivo biomedical imaging can be difficult to achieve. By scanning a line of illumination in one direction in conjunction with a stationary confocal slit aperture, very high image acquisition speeds can be achieved, but at the cost of a reduction in image quality. Here, the design, implementation, and experimental verification of a custom multi-point aperture modification to a line-scanning multi-spectral confocal microendoscope is presented. This new design improves the axial resolution of a line-scan system while maintaining high imaging rates. In addition, compared to the line-scanning configuration, previously reported simulations predicted that the multi-point aperture geometry greatly reduces the effects of tissue scatter on image quality. Experimental results confirming this prediction are presented.
Highlights
Confocal microscopy is an established technique for imaging thick biological specimens
Confocal microscopes operate by scanning an illumination pattern across an object
The aperture reduces the light throughput from out of focus planes in the sample, which is the basis for the optical sectioning property of confocal imaging systems
Summary
Confocal microscopy is an established technique for imaging thick biological specimens. The return signal is de-scanned and imaged to a conjugate plane that contains an aperture corresponding to the scanned illumination pattern. The aperture reduces the light throughput from out of focus planes in the sample, which is the basis for the optical sectioning property of confocal imaging systems. Confocal microscopes have been adapted for in vivo use by integrating them into portable instruments called confocal microendoscopes (or confocal endomicroscopes). Such systems are one of a class of techniques described as “optical biopsy” [1,2,3,4,5,6,7] that enable non-destructive in situ evaluation of tissue for real-time disease diagnosis
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