scholarly journals Content-Aware Image Restoration: Pushing the Limits of Fluorescence Microscopy

2017 ◽  
Author(s):  
Martin Weigert ◽  
Uwe Schmidt ◽  
Tobias Boothe ◽  
Andreas Müller ◽  
Alexandr Dibrov ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Image Restoration ◽  
Light Exposure ◽  
Axial Direction ◽  
Isotropic Resolution ◽  
Biological Phenomena ◽  
Trade Offs ◽  
Under Sampling ◽  
Key Driver ◽  
Photon Exposure

Fluorescence microscopy is a key driver of discoveries in the life-sciences, with observable phenomena being limited by the optics of the microscope, the chemistry of the fluorophores, and the maximum photon exposure tolerated by the sample. These limits necessitate trade-offs between imaging speed, spatial resolution, light exposure, and imaging depth. In this work we show how image restoration based on deep learning extends the range of biological phenomena observable by microscopy. On seven concrete examples we demonstrate how microscopy images can be restored even if 60-fold fewer photons are used during acquisition, how near isotropic resolution can be achieved with up to 10-fold under-sampling along the axial direction, and how tubular and granular structures smaller than the diffraction limit can be resolved at 20-times higher frame-rates compared to state-of-the-art methods. All developed image restoration methods are freely available as open source software in Python, Fiji, and Knime.

scholarly journals A versatile compressed sensing scheme for faster and less phototoxic 3D fluorescence microscopy

2017 ◽  
Author(s):  
Maxime Woringer ◽  
Xavier Darzacq ◽  
Christophe Zimmer ◽  
Mustafa Mir
Keyword(s):  
Fluorescence Microscopy ◽  
Compressed Sensing ◽  
Light Exposure ◽  
Three Dimensional ◽  
Light Sheet ◽  
Computational Imaging ◽  
Acquisition Time ◽  
Trade Offs ◽  
Fold Reduction ◽  
3D Fluorescence Microscopy

AbstractThree-dimensional fluorescence microscopy based on Nyquist sampling of focal planes faces harsh trade-offs between acquisition time, light exposure, and signal-to-noise. We propose a 3D compressed sensing approach that uses temporal modulation of the excitation intensity during axial stage sweeping and can be adapted to fluorescence microscopes without hardware modification. We describe implementations on a lattice light sheet microscope and an epifluorescence microscope, and show that images of beads and biological samples can be reconstructed with a 5-10 fold reduction of light exposure and acquisition time. Our scheme opens a new door towards faster and less damaging 3D fluorescence microscopy.OCIS codes: (110.1758) Computational imaging; (170.2520) Fluorescence microscopy; (170.6900) Three-dimensional microscopy.

scholarly journals Advanced Static and Dynamic Fluorescence Microscopy Techniques to Investigate Drug Delivery Systems

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 861
Author(s):  
Jacopo Cardellini ◽  
Arianna Balestri ◽  
Costanza Montis ◽  
Debora Berti
Keyword(s):  
Drug Delivery ◽  
Fluorescence Microscopy ◽  
Drug Delivery Systems ◽  
Laser Scanning ◽  
Super Resolution ◽  
Laser Scanning Confocal Microscopy ◽  
Delivery Systems ◽  
Scanning Confocal Microscopy ◽  
Biological Phenomena

In the past decade(s), fluorescence microscopy and laser scanning confocal microscopy (LSCM) have been widely employed to investigate biological and biomimetic systems for pharmaceutical applications, to determine the localization of drugs in tissues or entire organisms or the extent of their cellular uptake (in vitro). However, the diffraction limit of light, which limits the resolution to hundreds of nanometers, has for long time restricted the extent and quality of information and insight achievable through these techniques. The advent of super-resolution microscopic techniques, recognized with the 2014 Nobel prize in Chemistry, revolutionized the field thanks to the possibility to achieve nanometric resolution, i.e., the typical scale length of chemical and biological phenomena. Since then, fluorescence microscopy-related techniques have acquired renewed interest for the scientific community, both from the perspective of instrument/techniques development and from the perspective of the advanced scientific applications. In this contribution we will review the application of these techniques to the field of drug delivery, discussing how the latest advancements of static and dynamic methodologies have tremendously expanded the experimental opportunities for the characterization of drug delivery systems and for the understanding of their behaviour in biologically relevant environments.

scholarly journals Fourier ring correlation simplifies image restoration in fluorescence microscopy

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Sami Koho ◽  
Giorgio Tortarolo ◽  
Marco Castello ◽  
Takahiro Deguchi ◽  
Alberto Diaspro ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Image Restoration

scholarly journals Combined carbon photon and hydrogel therapy mediates the synergistic repair of full-thickness skin wounds

2020 ◽  
Vol 48 (8) ◽  
pp. 030006052093532
Author(s):  
Fan Yang ◽  
Xiuling Zhou ◽  
Sitong Chen ◽  
Qiuju Li ◽  
Ronghang Li ◽  
...  
Keyword(s):  
Wound Healing ◽  
Light Exposure ◽  
Tissue Formation ◽  
Full Thickness ◽  
Granulation Tissue Formation ◽  
Treatment Groups ◽  
Wound Model ◽  
Thickness Skin ◽  
Treatment Of Wounds ◽  
Photon Exposure

Objective This study investigated the synergistic repair effects of Prontosan hydrogel and carbon photon therapy in a rat full-thickness wound model. Methods The wavelength distribution of the photon source was determined. Dehydration of the Prontosan hydrogel and fibroblast viability were analyzed following exposure to different durations of light exposure at different distances from the source. Indexes of wound healing in a full-thickness rat wound model were then determined in groups ( n = 8 each) subjected to either no treatment, Prontosan treatment only, carbon photon therapy only, or a combination of the two treatments (synergistic group). Results Carbon photon exposure for 15 minutes at a distance of 20 cm from the wound was found to be optimal. Wound healing occurred faster in the synergistic group compared with the control and single-treatment groups. Growth factor secretion, granulation tissue formation, inflammation regulation, collagen deposition, and neovascularization were all higher in the synergistic group. Conclusions Prontosan hydrogel combined with carbon photon therapy may provide an optimal environment for wound healing and serve as a novel physical approach to the treatment of wounds. However, the number of animals included in this study was relatively small and a larger study is required to confirm these findings.

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