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Adhering interacting cells to two opposing coverslips allows super-resolution imaging of cell-cell interfaces.

ABSTRACT: Cell-cell interfaces convey mechanical and chemical information in multicellular systems. Microscopy has revealed intricate structure of such interfaces, yet typically with limited resolution due to diffraction and unfavourable orthogonal orientation of the interface to the coverslip. We present a simple and robust way to align cell-cell interfaces in parallel to the coverslip by adhering the interacting cells to two opposing coverslips. We demonstrate high-quality diffraction-limited and super-resolution imaging of interfaces (immune-synapses) between fixed and live CD8⁺ T-cells and either antigen presenting cells or melanoma cells. Imaging methods include bright-field, confocal, STED, dSTORM, SOFI, SRRF and large-scale tiled images. The low background, lack of aberrations and enhanced spatial stability of our method relative to existing cell-trapping techniques allow use of these methods. We expect that the simplicity and wide-compatibility of our approach will allow its wide dissemination for super-resolving the intricate structure and molecular organization in a variety of cell-cell interfaces.

SUBMITTER: Sajman J

PROVIDER: S-EPMC8016881 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature

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Adhering interacting cells to two opposing coverslips allows super-resolution imaging of cell-cell interfaces.

Sajman Julia J Razvag Yair Y Schidorsky Shachar S Kinrot Seon S Hermon Kobi K Yakovian Oren O Sherman Eilon E

Communications biology 20210401 1

Cell-cell interfaces convey mechanical and chemical information in multicellular systems. Microscopy has revealed intricate structure of such interfaces, yet typically with limited resolution due to diffraction and unfavourable orthogonal orientation of the interface to the coverslip. We present a simple and robust way to align cell-cell interfaces in parallel to the coverslip by adhering the interacting cells to two opposing coverslips. We demonstrate high-quality diffraction-limited and super- ...[more]

PMID: 33795833

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Project description:Super-resolution (SR) techniques are used in PET imaging to generate a high-resolution image by combining multiple low-resolution images that have been acquired from different points of view (POV). In this article, the authors propose a novel implementation of the SR technique whereby the required multiple low-resolution images are generated by shifting the reconstruction pixel grid during the image reconstruction process rather than being acquired from different POVs. The objective of this article is to compare the performances of the two SR implementations using theoretical and experimental studies. A mathematical framework is first provided to support the hypothesis that the two SR implementations have similar performance in current PET/CT scanners that use block detectors. Based on this framework, a simulation study, a point source study, and a NEMA/IEC phantom study were conducted to compare the performance of these two SR implementations with respect to contrast, resolution, noise, and SNR. For reference purposes, a comparison with a native reconstruction (NR) image using a high-resolution pixel grid was also performed. The mathematical framework showed that the two SR implementations are expected to achieve similar contrast and resolution but different noise contents. These results were confirmed by the simulation and experimental studies. The simulation study showed that the two SR implementations have an average contrast difference of 2.3%, while the point source study showed that their average differences in contrast and resolution were 0.5% and 1.2%, respectively. Comparisons between the SR and NR images for the point source study showed that the NR image exhibited averages of 30% and 8% lower contrast and resolution, respectively. The NEMA/IEC phantom study showed that the three images (two SR and NR) exhibited different noise structures. The SNR of the new SR implementation was, on average, 21.5% lower than the original implementation largely due to an increase in background noise, while the NR image had averages of 18.5% and 8% lower SNR and contrast, respectively, versus the two SR images. The new SR implementation can potentially replace the original SR approach in current PET scanners that use block detectors while maintaining similar contrast and resolution, but at a relatively lower SNR. A major advantage of the new SR implementation is its shorter overall scan duration which results in an increase in scanner throughput and a reduction in patient motion.

Dataset Information

Adhering interacting cells to two opposing coverslips allows super-resolution imaging of cell-cell interfaces.

Publications

Adhering interacting cells to two opposing coverslips allows super-resolution imaging of cell-cell interfaces.

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