Share Email Print

Journal of Biomedical Optics

Superior performance with sCMOS over EMCCD in super-resolution optical fluctuation imaging
Author(s): Xuanze Chen; Zhiping Zeng; Rongqin Li; Boxin Xue; Peng Xi; Yujie Sun
Format Member Price Non-Member Price
PDF $20.00 $25.00
cover GOOD NEWS! Your organization subscribes to the SPIE Digital Library. You may be able to download this paper for free. Check Access

Paper Abstract

Super-resolution optical fluctuation imaging (SOFI) is a fast and low-cost live-cell optical nanoscopy for extracting subdiffraction information from the statistics of fluorescence intensity fluctuation. As SOFI is based on the fluctuation statistics, rather than the detection of single molecules, it poses unique requirements for imaging detectors, which still lack a systematic evaluation. Here, we analyze the influences of pixel sizes, frame rates, noise levels, and different gains in SOFI with simulations and experimental tests. Our analysis shows that the smaller pixel size and faster readout speed of scientific-grade complementary metal oxide semiconductor (sCMOS) enables SOFI to achieve high spatiotemporal resolution with a large field-of-view, which is especially beneficial for live-cell super-resolution imaging. Overall, as the performance of SOFI is relatively insensitive to the signal-to-noise ratio (SNR), the gain in pixel size and readout speed exceeds the loss in SNR, indicating sCMOS is superior to electron multiplying charge coupled device in context to SOFI in many cases. Super-resolution imaging of cellular microtubule structures with high-order SOFI is experimentally demonstrated at large field-of-view, taking advantage of the large pixel number and fast frame rate of sCMOS cameras.

Paper Details

Date Published: 9 June 2016
PDF: 10 pages
J. Biomed. Opt. 21(6) 066007 doi: 10.1117/1.JBO.21.6.066007
Published in: Journal of Biomedical Optics Volume 21, Issue 6
Show Author Affiliations
Xuanze Chen, Peking Univ. (China)
Zhiping Zeng, Peking Univ. (China)
Rongqin Li, Peking Univ. (China)
Boxin Xue, Peking Univ. (China)
Peng Xi, Peking Univ. (China)
Yujie Sun, Peking Univ. (China)

© SPIE. Terms of Use
Back to Top