Nanosheet wrapping-assisted coverslip-free imaging for looking deeper into a tissue at high resolution


Autoři: Hong Zhang aff001;  Kenji Yarinome aff003;  Ryosuke Kawakami aff004;  Kohei Otomo aff004;  Tomomi Nemoto aff004;  Yosuke Okamura aff001
Působiště autorů: Department of Applied Chemistry, School of Engineering, Tokai University, Kanagawa, Japan aff001;  Micro/Nano Technology Center, Tokai University, Kanagawa, Japan aff002;  Course of Applied Science, Graduate School of Engineering, Tokai University, Kanagawa, Japan aff003;  Research Institute for Electronic Science, Hokkaido University, Sapporo, Japan aff004;  Graduate School of Information Science and Technology, Hokkaido University, Sapporo, Japan aff005;  Department of Molecular Medicine for Pathogenesis, Ehime University Graduate School of Medicine, Ehime, Japan aff006;  Exploratory Research Center on Life and Living Systems, National Institute of Natural Sciences, Aichi, Japan aff007;  National Institute for Physiological Sciences, Aichi, Japan aff008;  The Graduate University for Advanced Studies (SOKENDAI), Aichi, Japan aff009
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.pone.0227650

Souhrn

In order to achieve deep tissue imaging, a number of optical clearing agents have been developed. However, in a conventional microscopy setup, an objective lens can only be moved until it is in contact with a coverslip, which restricts the maximum focusing depth into a cleared tissue specimen. Until now, it is still a fact that the working distance of a high magnification objective lens with a high numerical aperture is always about 100 μm. In this study, a polymer thin film (also called as nanosheet) composed of fluoropolymer with a thickness of 130 nm, less than one-thousandth that of a 170 μm thick coverslip, is employed to replace the coverslip. Owing to its excellent characteristics, such as high optical transparency, mechanical robustness, chemical resistance, and water retention ability, nanosheet is uniquely capable of providing a coverslip-free imaging. By wrapping the tissue specimen with a nanosheet, an extra distance of 170 μm for the movement of objective lens is obtained. Results show an equivalently high resolution imaging can be obtained if a homogenous refractive index between immersion liquid and mounting media is adjusted. This method will facilitate a variety of imaging tasks with off-the-shelf high magnification objectives.

Klíčová slova:

Fluorescence imaging – Gel electrophoresis – Imaging techniques – Neuroimaging – Oils – Optical lenses – Thin films – Nanowires


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