Edge roughness analysis in nanoscale for single-molecule localization microscopy images
Author:
Jeong Uidon1ORCID, Go Ga-eun1, Jeong Dokyung1, Lee Dongmin1, Kim Min Jeong1, Kang Minjae1, Kim Namyoon2, Jung Jaehwang2, Kim Wookrae2, Lee Myungjun2, Kim Doory13ORCID
Affiliation:
1. Department of Chemistry , Hanyang University , Seoul 04763 , Republic of Korea 2. MI Equipment R&D Team, Mechatronics Research, Samsung Electronics Co., Ltd. , Hwaseong 18848 , Republic of Korea 3. Research Institute for Convergence of Basic Sciences, Institute of Nano Science and Technology, and Research Institute for Natural Sciences, Hanyang University , Seoul 04763 , Republic of Korea
Abstract
Abstract
The recent advances in super-resolution fluorescence microscopy, including single-molecule localization microscopy (SMLM), has enabled the study of previously inaccessible details, such as the organization of proteins within cellular compartments and even nanostructures in nonbiological nanomaterials, such as the polymers and semiconductors. With such developments, the need for the development of various computational nanostructure analysis methods for SMLM images is also increasing; however, this has been limited to protein cluster analysis. In this study, we developed an edge structure analysis method for pointillistic SMLM images based on the line edge roughness and power spectral density analyses. By investigating the effect of point properties in SMLM images, such as the size, density, and localization precision on the roughness measurement, we successfully demonstrated this analysis method for experimental SMLM images of actual samples, including the semiconductor line patterns, cytoskeletal elements, and cell membranes. This systematic investigation of the effect of each localization rendering parameter on edge roughness measurement provides a range for the optimal rendering parameters that preserve the relevant nanoscale structure of interest. These new methods are expected to expand our understanding of the targets by providing valuable insights into edge nanoscale structures that have not been previously obtained quantitatively.
Funder
Samsung National Research Foundation of Korea
Publisher
Walter de Gruyter GmbH
Subject
Electrical and Electronic Engineering,Atomic and Molecular Physics, and Optics,Electronic, Optical and Magnetic Materials,Biotechnology
Reference40 articles.
1. S. W. Hell and J. Wichmann, “Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy,” Opt. Lett., vol. 19, no. 11, pp. 780–782, 1994. https://doi.org/10.1364/ol.19.000780. 2. M. G. Gustafsson, D. A. Agard, and J. W. Sedat, “Sevenfold improvement of axial resolution in 3D wide-field microscopy using two objective-lenses,” in Three-dimensional microscopy: image acquisition and processing II, vol. 2412, San Jose, CA, USA, SPIE, 1995, pp. 147–156. 3. M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods, vol. 3, no. 10, pp. 793–796, 2006. https://doi.org/10.1038/nmeth929. 4. E. Betzig, et al.., “Imaging intracellular fluorescent proteins at nanometer resolution,” Science, vol. 313, no. 5793, pp. 1642–1645, 2006. https://doi.org/10.1126/science.1127344. 5. J. Lippincott-Schwartz and S. Manley, “Putting super-resolution fluorescence microscopy to work,” Nat. Methods, vol. 6, no. 1, pp. 21–23, 2009. https://doi.org/10.1038/nmeth.f.233.
|
|