Multimodal on-chip nanoscopy and quantitative phase imaging reveals the nanoscale morphology of liver sinusoidal endothelial cells.

Autor:	Butola A; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway.; Bio-photonics and Green Photonics Laboratory, Department of Physics, Indian Institute of Technology Delhi, New Delhi 110016, India., Coucheron DA; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway., Szafranska K; Faculty of Health Sciences, Department of Medical Biology, Vascular Biology Research Group, UiT The Arctic University of Norway, Tromsø 9037, Norway., Ahmad A; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway., Mao H; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway., Tinguely JC; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway., McCourt P; Faculty of Health Sciences, Department of Medical Biology, Vascular Biology Research Group, UiT The Arctic University of Norway, Tromsø 9037, Norway., Senthilkumaran P; Bio-photonics and Green Photonics Laboratory, Department of Physics, Indian Institute of Technology Delhi, New Delhi 110016, India., Mehta DS; Bio-photonics and Green Photonics Laboratory, Department of Physics, Indian Institute of Technology Delhi, New Delhi 110016, India., Agarwal K; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway., Ahluwalia BS; Department of Physics and Technology, Universitetet i Tromsø (UiT) The Arctic University of Norway, 9037 Tromsø, Norway; balpreet.singh.ahluwalia@uit.no.; Department of Clinical Science, Intervention and Technology, Karolinska Institutet, 17177 Stockholm, Sweden.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2021 Nov 23; Vol. 118 (47).
DOI:	10.1073/pnas.2115323118
Abstrakt:	Visualization of three-dimensional (3D) morphological changes in the subcellular structures of a biological specimen is a major challenge in life science. Here, we present an integrated chip-based optical nanoscopy combined with quantitative phase microscopy (QPM) to obtain 3D morphology of liver sinusoidal endothelial cells (LSEC). LSEC have unique morphology with small nanopores (50-300 nm in diameter) in the plasma membrane, called fenestrations. The fenestrations are grouped in discrete clusters, which are around 100 to 200 nm thick. Thus, imaging and quantification of fenestrations and sieve plate thickness require resolution and sensitivity of sub-100 nm along both the lateral and the axial directions, respectively. In chip-based nanoscopy, the optical waveguides are used both for hosting and illuminating the sample. The fluorescence signal is captured by an upright microscope, which is converted into a Linnik-type interferometer to sequentially acquire both superresolved images and phase information of the sample. The multimodal microscope provided an estimate of the fenestration diameter of 119 ± 53 nm and average thickness of the sieve plates of 136.6 ± 42.4 nm, assuming the constant refractive index of cell membrane to be 1.38. Further, LSEC were treated with cytochalasin B to demonstrate the possibility of precise detection in the cell height. The mean phase value of the fenestrated area in normal and treated cells was found to be 161 ± 50 mrad and 109 ± 49 mrad, respectively. The proposed multimodal technique offers nanoscale visualization of both the lateral size and the thickness map, which would be of broader interest in the fields of cell biology and bioimaging. Competing Interests: Competing interest statement: B.S.A. has applied for patent GB1606268.9 for chip-based optical nanoscopy. B.S.A. is a cofounder of the company Chip NanoImaging AS, which commercializes on-chip superresolution microscopy systems. (Copyright © 2021 the Author(s). Published by PNAS.)
Databáze:	MEDLINE
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