Mahdi Mazaheri, Kiarash Kasaian, David Albrecht, Jan Renger, Tobias Utikal, Cornelia Holler, Vahid Sandoghdar
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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.
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.