iSCAT 显微镜和具有定制空间相干性的粒子追踪技术

IF 8.4 1区 物理与天体物理 Q1 OPTICS
Optica Pub Date : 2024-06-21 DOI:10.1364/optica.523788
Mahdi Mazaheri, Kiarash Kasaian, David Albrecht, Jan Renger, Tobias Utikal, Cornelia Holler, Vahid Sandoghdar
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引用次数: 0

摘要

干涉散射(iSCAT)显微镜在检测孤立纳米粒子和分子并对其成像的无标记光学方法中表现出无与伦比的性能。然而,当对生物细胞等复杂结构成像时,来自样品不同位置的散射场的叠加会导致斑点状背景,这对破译精细特征构成了巨大挑战。在这里,我们展示了通过控制照明的空间相干性,可以在不牺牲灵敏度的情况下消除虚假斑点。我们通过在照明路径中放置一个旋转扩散器、一个可调透镜和一个光圈来演示这种方法。我们报告了在保持衍射极限分辨率的同时,以 25 kHz 的高帧频和 100µm×100µm 的大视场进行成像的情况。我们通过对单个 COS-7 细胞中的 1000 多个囊泡进行三维(3D)跟踪,以及对内质网(ER)网络的动态成像,展示了这些功能的优势。我们的方法为使用宽视场 iSCAT 显微镜将无标记成像、灵敏检测和三维高速跟踪相结合打开了大门。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
iSCAT microscopy and particle tracking with tailored spatial coherence
Interferometric scattering (iSCAT) microscopy has demonstrated unparalleled performance among label-free optical methods for detecting and imaging isolated nanoparticles and molecules. However, when imaging complex structures such as biological cells, the superposition of the scattering fields from different locations of the sample leads to a speckle-like background, posing a significant challenge in deciphering fine features. Here, we show that by controlling the spatial coherence of the illumination, one can eliminate the spurious speckle without sacrificing sensitivity. We demonstrate this approach by positioning a rotating diffuser coupled with an adjustable lens and an iris in the illumination path. We report on imaging at a high frame rate of 25 kHz and across a large field of view of 100µm×100µm, while maintaining diffraction-limited resolution. We showcase the advantages of these features by three-dimensional (3D) tracking over 1000 vesicles in a single COS-7 cell and by imaging the dynamics of the endoplasmic reticulum (ER) network. Our approach opens the door to the combination of label-free imaging, sensitive detection, and 3D high-speed tracking using wide-field iSCAT microscopy.
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来源期刊
Optica
Optica OPTICS-
CiteScore
19.70
自引率
2.90%
发文量
191
审稿时长
2 months
期刊介绍: Optica is an open access, online-only journal published monthly by Optica Publishing Group. It is dedicated to the rapid dissemination of high-impact peer-reviewed research in the field of optics and photonics. The journal provides a forum for theoretical or experimental, fundamental or applied research to be swiftly accessed by the international community. Optica is abstracted and indexed in Chemical Abstracts Service, Current Contents/Physical, Chemical & Earth Sciences, and Science Citation Index Expanded.
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