ORCID Profile
0000-0002-1260-8262
Current Organisations
UNSW Sydney
,
NetTargets, Inc.
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Publisher: Springer Science and Business Media LLC
Date: 03-02-2022
DOI: 10.1038/S41467-022-28219-6
Abstract: Two-photon direct laser writing is an additive fabrication process that utilizes two-photon absorption of tightly focused femtosecond laser pulses to implement spatially controlled polymerization of a liquid-phase photoresist. Two-photon direct laser writing is capable of nanofabricating arbitrary three-dimensional structures with nanometer accuracy. Here, we explore direct laser writing for high-resolution optical microscopy by fabricating unique 3D optical fiducials for single-molecule tracking and 3D single-molecule localization microscopy. By having control over the position and three-dimensional architecture of the fiducials, we improve axial discrimination and demonstrate isotropic subnanometer 3D focusing ( .8 nm) over tens of micrometers using a standard inverted microscope. We perform 3D single-molecule acquisitions over cellular volumes, unsupervised data acquisition and live-cell single-particle tracking with nanometer accuracy.
Publisher: American Association for the Advancement of Science (AAAS)
Date: 17-04-2020
Abstract: Feedback SMLM achieves ultrahigh localization precision and in situ distance measurements on the biological relevant scales.
Publisher: Bio-Protocol, LLC
Date: 2021
Publisher: Elsevier BV
Date: 10-2020
Publisher: SPIE
Date: 29-04-2017
DOI: 10.1117/12.2275706
Publisher: American Chemical Society (ACS)
Date: 04-03-2020
DOI: 10.1021/JACS.0C00411
Publisher: Springer Science and Business Media LLC
Date: 02-12-2021
DOI: 10.1038/S41596-020-00426-9
Abstract: A key part of any super-resolution technique involves accurately correcting for mechanical motion of the s le and setup during acquisition. If left uncorrected, drift degrades the resolution of the final reconstructed image and can introduce unwanted artifacts. Here, we describe how to implement active stabilization, thereby reducing drift to ~1 nm across all three dimensions. In this protocol, we show how to implement our method on custom and standard microscopy hardware. We detail the construction of a separate illumination and detection path, dedicated exclusively to acquiring the diffraction pattern of fiducials deposited on the imaging slide. We also show how to focus lock and adjust the focus in arbitrary nanometer step size increments. Our real-time focus locking is based on kHz calculations performed using the graphics processing unit. The fast calculations allow for rapid repositioning of the s le, which reduces drift below the photon-limited localization precision. Our approach allows for a single-molecule and/or super-resolution image acquisition free from movement artifacts and eliminates the need for complex algorithms or hardware installations. The method is also useful for long acquisitions which span over hours or days, such as multicolor super resolution. Installation does not require specialist knowledge and can be implemented in standard biological laboratories. The full protocol can be implemented within ~2 weeks.
No related grants have been discovered for Jongho Baek.