2022-12-06 ワシントン大学セントルイス校
宇宙望遠鏡と同様に、半径方向および方位角方向に偏光した多視点反射鏡（raMVR）顕微鏡は、できるだけ多くの光を集めることに依存している。しかし、その光は遠くのものを見るために使うのではなく、タンパク質や細胞膜に付着した小さな蛍光分子のさまざまな特徴を見分けるために使うのである。
raMVR顕微鏡は、波長板と呼ばれる偏光光学系とピラミッド型のミラーを使って光を8つのチャンネルに分離し、それぞれのチャンネルが分子の位置と向きの異なる部分を表している。
＜関連情報＞
- https://source.wustl.edu/2022/12/telescope-inspired-microscope-sees-molecules-in-6d/
- https://www.nature.com/articles/s41566-022-01116-6
マルチビューリフレクター顕微鏡による等方性分解能の6次元単一分子イメージング Six-dimensional single-molecule imaging with isotropic resolution using a multi-view reflector microscope
Oumeng Zhang,Zijian Guo,Yuanyuan He,Tingting Wu,Michael D. Vahey & Matthew D. Lew
Nature Photonics Published;05 December 2022
DOI:https://doi.org/10.1038/s41566-022-01116-6
Abstract
Imaging of both the positions and orientations of single fluorophores, termed single-molecule orientation-localization microscopy, is a powerful tool for the study of biochemical processes. However, the limited photon budget associated with single-molecule fluorescence makes high-dimensional imaging with isotropic, nanoscale spatial resolution a formidable challenge. Here we realize a radially and azimuthally polarized multi-view reflector (raMVR) microscope for the imaging of the three-dimensional (3D) positions and 3D orientations of single molecules, with precisions of 10.9 nm and 2.0° over a 1.5-μm depth range. The raMVR microscope achieves 6D super-resolution imaging of Nile red molecules transiently bound to lipid-coated spheres, accurately resolving their spherical morphology, despite refractive-index mismatch. By observing the rotational dynamics of Nile red, raMVR images also resolve the infiltration of lipid membranes by amyloid-beta oligomers without covalent labelling. Finally, we demonstrate 6D imaging of cell membranes, where the orientations of specific fluorophores reveal heterogeneity in membrane fluidity. With its nearly isotropic 3D spatial resolution and orientation measurement precision, we expect the raMVR microscope to enable 6D imaging of molecular dynamics within biological and chemical systems with exceptional detail.
