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Proceedings Article

Multi-color quantum dot stochastic optical reconstruction microscopy (qSTORM)

[+] Author Affiliations
Kayvan F. Tehrani, Jianquan Xu, Peter A. Kner

The Univ. of Georgia (United States)

Proc. SPIE 9331, Single Molecule Spectroscopy and Superresolution Imaging VIII, 93310C (March 9, 2015); doi:10.1117/12.2077293
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From Conference Volume 9331

  • Single Molecule Spectroscopy and Superresolution Imaging VIII
  • Jörg Enderlein; Ingo Gregor; Zygmunt K. Gryczynski; Rainer Erdmann; Felix Koberling
  • San Francisco, California, United States | February 07, 2015

abstract

Although Single Molecule Localization (SML) techniques have pushed the resolution of fluorescence microscopy beyond the diffraction limit, the accuracy of SML has been limited by the brightness of the fluorophores. The introduction of Quantum Dots (QD) for SML promises to overcome this barrier, and the QD Blueing technique provides a novel approach to SML microscopy. QDs have a higher quantum yield and absorption cross-section, making them brighter, thereby providing a higher accuracy of localization. The blueing technique is also faster and more quantitative than other SML techniques such as dSTORM. The initial bleaching step required by dSTORM is not necessary and each QD is imaged only once as its emission spectrum moves through the blueing window in contrast to dSTORM where the same molecule might be imaged multiple times. Single color QD Blueing has been demonstrated. However in biological imaging, multi-color imaging is essential for understanding the samples under study. Here we introduce two color superresolution microscopy using QD Blueing on biological samples. We demonstrate simultaneous imaging of microtubules and mitochondria in HepG2 cells with a localization accuracy of 40nm. We further show how QD Blueing can be optimized through the control of the sample mounting medium. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Citation

Kayvan F. Tehrani ; Jianquan Xu and Peter A. Kner
" Multi-color quantum dot stochastic optical reconstruction microscopy (qSTORM) ", Proc. SPIE 9331, Single Molecule Spectroscopy and Superresolution Imaging VIII, 93310C (March 9, 2015); doi:10.1117/12.2077293; http://dx.doi.org/10.1117/12.2077293


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