Simple differential digital confocal aperture to improve axial response of line-scanning confocal microendoscopes

Tang, Yubo; Kortum, Alex; Vohra, Imran S.; Carns, Jennifer; Anandasabapathy, Sharmila; Richards‐Kortum, Rebecca

doi:10.1364/ol.44.004519

“…These improvements often limit their application scenarios and prevent their widespread application in biomedical research. Unlike previous difference methods [30][31][32] using multi-scanning or multi-detector, our method constructs the different PSFs integrating TDI technology and redundant information on multi-line detection in a single scan.…”

Section: Discussionmentioning

confidence: 99%

Differential synthetic illumination based on multi-line detection for resolution and contrast enhancement of line confocal microscopy

Yuan¹,

Wang²,

Li³

et al. 2023

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Line confocal (LC) microscopy is a fast 3D imaging technique, but its asymmetric detection slit limits resolution and optical sectioning. To address this, we propose the differential synthetic illumination (DSI) method based on multi-line detection to enhance the spatial resolution and optical sectioning capability of the LC system. The DSI method allows the imaging process to simultaneously accomplish on a single camera, which ensures the rapidity and stability of the imaging process. DSI-LC improves X- and Z-axis resolution by 1.28 and 1.26 times, respectively, and optical sectioning by 2.6 times compared to LC. Furthermore, the spatially resolved power and contrast are also demonstrated by imaging pollen, microtubule, and the fiber of the GFP fluorescence-labeled mouse brain. Finally, Video-rate imaging of zebrafish larval heart beating in a 665.6×332.8μm² field-of-view is achieved. DSI-LC provides a promising approach for 3D large-scale and functional imaging in vivo with improved resolution, contrast, and robustness.

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“…To address this issue, we introduce a difference method that has been employed in various imaging modes to improve spatial resolution and imaging contrast due to its simplicity [26][27][28][29]. Recently, the difference methods had been used to improve LC performance, but these methods used multiple scans or multiple detectors to obtain different point spread functions (PFS) [30][31][32]. The multi-scanning acquisition would reduce imaging speed.…”

Section: Introductionmentioning

confidence: 99%

Differential synthetic illumination based on multi-line detection for resolution and contrast enhancement of line confocal microscopy

Yuan¹,

Wang²,

Li³

et al. 2023

Preprint

0

View full text Add to dashboard Cite

Line confocal (LC) microscopy is a fast 3D imaging technique, but its asymmetric detection slit limits resolution and optical sectioning. To address this, we propose the differential synthetic illumination (DSI) method based on multi-line detection to enhance the spatial resolution and optical sectioning capability of the LC system. The DSI method allows the imaging process to simultaneously accomplish on a single camera, which ensures the rapidity and stability of the imaging process. DSI-LC improves X- and Z-axis resolution by 1.28 and 1.26 times, respectively, and optical sectioning by 2.6 times compared to LC. Furthermore, the spatially resolved power and contrast are also demonstrated by imaging pollen, microtubule, and the fiber of the GFP fluorescence-labeled mouse brain. Finally, Video-rate imaging of zebrafish larval heart beating in a 665.6×332.8μm² field-of-view is achieved. DSI-LC provides a promising approach for 3D large-scale and functional imaging in vivo with improved resolution, contrast, and robustness.

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“…A two-camera approach was recently proposed, which avoids these inter-frame motion artefacts. 19 Rather than acquiring the two images sequentially, the beam is split between two rolling shutter cameras, one with triggering timing adjusted to provide an aligned-slit (the confocal image) and one providing an offset-slit (the residual image). The authors used an LED source to generate the excitation line, with the scanning line created by displaying a series of patterns on a digital micromirror device (DMD), a programmable array of mirrors.…”

Section: Introductionmentioning

confidence: 99%

Reduced motion artifacts and speed improvements in enhanced line-scanning fiber bundle endomicroscopy

Thrapp

¹

,

Hughes

²

2021

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Significance: Confocal laser scanning enables optical sectioning in fiber bundle endomicroscopy but limits the frame rate. To be able to better explore tissue morphology, it is useful to stitch sequentially acquired frames into a mosaic. However, low frame rates limit the maximum probe translation speed. Line-scanning (LS) confocal endomicroscopy provides higher frame rates, but residual out-of-focus light degrades images. Subtraction-based approaches can suppress this residue at the expense of introducing motion artifacts.Aim: To generate high-frame-rate endomicroscopy images with improved optical sectioning, we develop a high-speed subtraction method that only requires the acquisition of a single camera frame.Approach: The rolling shutter of a CMOS camera acts as both the aligned and offset detector slits required for subtraction-based sectioning enhancement. Two images of the bundle are formed on different regions of the camera, allowing both images to be acquired simultaneously.Results: We confirm improved optical sectioning compared to conventional LS, particularly far from focus, and show that motion artifacts are not introduced. We demonstrate high-speed mosaicing at frame rates of up to 240 Hz. Conclusion:High-speed acquisition of optically sectioned images using the new subtraction based-approach leads to improved mosaicing at high frame rates.

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Simple differential digital confocal aperture to improve axial response of line-scanning confocal microendoscopes

Cited by 9 publications

References 14 publications

Differential synthetic illumination based on multi-line detection for resolution and contrast enhancement of line confocal microscopy

Differential synthetic illumination based on multi-line detection for resolution and contrast enhancement of line confocal microscopy

Differential synthetic illumination based on multi-line detection for resolution and contrast enhancement of line confocal microscopy

Reduced motion artifacts and speed improvements in enhanced line-scanning fiber bundle endomicroscopy

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