Single-shot optical sectioning microscopy based on structured illumination

2022 ◽  
Author(s):  
Zhiqiang Fu ◽  
Jialong Chen ◽  
Gan Liu ◽  
Shih-Chi CHEN
Keyword(s):  
Single Shot ◽  
Structured Illumination ◽  
Optical Sectioning

scholarly journals Artifact-free whole-slide imaging with structured illumination microscopy and Bayesian image reconstruction

GigaScience ◽  
2020 ◽  
Vol 9 (4) ◽  
Author(s):  
Karl A Johnson ◽  
Guy M Hagen
Keyword(s):  
Super Resolution ◽  
Structured Illumination ◽  
Optical Sectioning ◽  
Structured Illumination Microscopy ◽  
High Quality ◽  
Related Data ◽  
Reconstruction Methods ◽  
Set Up ◽  
Bayesian Image Reconstruction ◽  
Hematoxylin Eosin

Abstract Background Structured illumination microscopy (SIM) is a method that can be used to image biological samples and can achieve both optical sectioning and super-resolution effects. Optimization of the imaging set-up and data-processing methods results in high-quality images without artifacts due to mosaicking or due to the use of SIM methods. Reconstruction methods based on Bayesian estimation can be used to produce images with a resolution beyond that dictated by the optical system. Findings Five complete datasets are presented including large panoramic SIM images of human tissues in pathophysiological conditions. Cancers of the prostate, skin, ovary, and breast, as well as tuberculosis of the lung, were imaged using SIM. The samples are available commercially and are standard histological preparations stained with hematoxylin-eosin. Conclusion The use of fluorescence microscopy is increasing in histopathology. There is a need for methods that reduce artifacts caused by the use of image-stitching methods or optical sectioning methods such as SIM. Stitched SIM images produce results that may be useful for intraoperative histology. Releasing high-quality, full-slide images and related data will aid researchers in furthering the field of fluorescent histopathology.

scholarly journals Differential structured illumination microendoscopy for in vivo imaging of molecular contrast agents

2016 ◽  
Vol 113 (39) ◽  
pp. 10769-10773 ◽  
Author(s):  
Pelham Keahey ◽  
Preetha Ramalingam ◽  
Kathleen Schmeler ◽  
Rebecca R. Richards-Kortum
Keyword(s):  
Contrast Agents ◽  
In Vivo Imaging ◽  
Fiber Optic ◽  
Adenocarcinoma In Situ ◽  
Structured Illumination ◽  
Optical Sectioning ◽  
Optical Elements ◽  
Fiber Optic System ◽  
Molecular Contrast

Fiber optic microendoscopy has shown promise for visualization of molecular contrast agents used to study disease in vivo. However, fiber optic microendoscopes have limited optical sectioning capability, and image contrast is limited by out-of-focus light generated in highly scattering tissue. Optical sectioning techniques have been used in microendoscopes to remove out-of-focus light but reduce imaging speed or rely on bulky optical elements that prevent in vivo imaging. Here, we present differential structured illumination microendoscopy (DSIMe), a fiber optic system that can perform structured illumination in real time for optical sectioning without any opto-mechanical components attached to the distal tip of the fiber bundle. We demonstrate the use of DSIMe during in vivo fluorescence imaging in patients undergoing surgery for cervical adenocarcinoma in situ. Images acquired using DSIMe show greater contrast than standard microendoscopy, improving the ability to detect cellular atypia associated with neoplasia.

scholarly journals Fast, Accurate Polarization and Polarity Imaging with Polarized Structured Illumination

2019 ◽  
Author(s):  
Karl Zhanghao ◽  
Wenhui Liu ◽  
Meiqi Li ◽  
Xingye Chen ◽  
Chunyan Shan ◽  
...  
Keyword(s):  
Live Cells ◽  
Polarization Modulation ◽  
Optical Transfer Function ◽  
Wide Field ◽  
Structured Illumination ◽  
Optical Sectioning ◽  
Polarization Imaging ◽  
Order Of Magnitude ◽  
Ratiometric Imaging ◽  
Optical Transfer

AbstractThe orientation and wobbling behavior of the fluorescent dipoles are of great significance in revealing the structure and state of cells. Due to the poor optical sectioning capability of wide-field microscopy, the polarization modulation signals are susceptible to the neighboring fluorophores. The missing cone of wide field optical transfer function induces vast out-of-focus background, resulting in biased polarization orientation and decrease polarization factor. Here, we apply polarized structured illumination to achieve polarization modulation imaging with optical sectioning, and simultaneously measure the lipid polarity with two-color ratiometric imaging. Our results demonstrate a significant increase in measurement accuracy of not only the dipole orientations but also the wobbling behavior of the ensemble dipole. Compared to the conventional confocal polarization imaging, our method obtains an order-of-magnitude faster imaging speed, capturing the fast dynamics of subcellular structures in live cells.

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