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Spectroscopic super-resolution fluorescence cell imaging using ultra-small Ge quantum dots

Song, Mingying, Karatutlu, Ali, Ali, Isma, Ersoy, Osman, Zhou, Yun, Yang, Yongxin, Zhang, Yuanpeng, Little, William R., Wheeler, Ann P. and Sapelkin, Andrei V. (2017) Spectroscopic super-resolution fluorescence cell imaging using ultra-small Ge quantum dots Optics Express, 25 (4). pp. 4240-4253.

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We demonstrate a spectroscopic imaging based super-resolution approach by separating the overlapping diffraction spots into several detectors during a single scanning period and taking advantage of the size-dependent emission wavelength in nanoparticles. This approach has been tested using off-the-shelf quantum dots (Invitrogen Qdot) and inhouse novel ultra-small (�3 nm) Ge QDs. Furthermore, we developed a method-specific Gaussian fitting and maximum likelihood estimation based on a Matlab algorithm for fast QD localisation. This methodology results in a three-fold improvement in the number of localised QDs compared to non-spectroscopic images. With the addition of advanced ultra-small Ge probes, the number can be improved even further, giving at least 1.5 times improvement when compared to Qdots. Using a standard scanning confocal microscope we achieved a data acquisition rate of 200 ms per image frame. This is an improvement on single molecule localisation super-resolution microscopy where repeated image capture limits the imaging speed, and the size of fluorescence probes limits the possible theoretical localisation resolution. We show that our spectral deconvolution approach has a potential to deliver data acquisition rates on the ms scale thus providing super-resolution in live systems. © 2017, OSA - The Optical Society. All rights reserved.

Item Type: Article
Divisions : Faculty of Engineering and Physical Sciences > Electronic Engineering
Authors :
Song, Mingying
Karatutlu, Ali
Ali, Isma
Ersoy, Osman
Zhou, Yun
Zhang, Yuanpeng
Little, William R.
Wheeler, Ann P.
Sapelkin, Andrei V.
Date : 16 February 2017
DOI : 10.1364/OE.25.004240
Copyright Disclaimer : Copyright 2017 Optical Express Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.
Uncontrolled Keywords : Data acquisition, Fluorescence, Germanium, Maximum likelihood, Maximum likelihood estimation, Nanocrystals, Optical resolving power, Probes, Diffraction spots, Emission wavelength, Fluorescence probes, Scanning confocal microscopes, Spectral deconvolution, Spectroscopic images, Spectroscopic imaging, Super-resolution microscopy, Semiconductor quantum dots, quantum dot, fluorescence, spectrofluorometry, statistical model, Fluorescence, Likelihood Functions, Quantum Dots, Spectrometry, Fluorescence
Depositing User : Diane Maxfield
Date Deposited : 16 Apr 2019 15:00
Last Modified : 16 Apr 2019 15:00

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