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Abstract
We recently reported on the use of defocused imaging for single molecule localization microscopy close to the coverslip [Biomed. Opt. Express , 11, 775 (2020) [CrossRef] ]. The original manuscript contains an error in the description of the Cramér-Rao lower bound (CRLB) calculations for SALM/DONALD, which affected the corresponding precision plot in Figure 2(b). We present a corrected description and plot.
© 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
In a recent publication we proposed defocused imaging for single molecule localization microscopy (SMLM) close to the coverslip [1]. The description for calculating CRLB-based precision curves for the photometric approach used in SALM/DONALD provided in the appendix contains an error: For SALM/DONALD, the Fisher matrices for the PSFs of the two detection channels (UAF and TOT) should not contain z-derivatives, i.e., the last row and column in the Fisher matrix of Eq. (7) of the paper should not be included. This reflects the assumption that the z position of the molecule does not affect the signal estimates.
The resulting z-precision curve for SALM/DONALD shows improved precisions compared the plot in Figure 2(b) of our manuscript. The corrected curve is shown in Fig. 1 and compared to the z-precision curves of SALM/DONALD+, off-focus imaging ($\Gamma$ = 500 nm) and results from Monte Carlo (MC) simulations for each method (marked by dots, each dot marks the average of 300 estimates).
Fig. 1. Calculated localization precisions for SALM/DONALD, SALM/DONALD+ and off-focus imaging. The dots mark the results from Monte-Carlo simulations. (signal = 2000 photons, background level = 100 photons per pixel, wavelength = 670 nm, pixel size = 115 nm, image size = 15$\times$15 pixel, NA = 1.7)
Acknowledgements
We would like to thank Jonas Ries for a helpful discussion about CRLB calculations.
References
1. P. Zelger, L. Bodner, L. Velas, G. J. Schütz, and A. Jesacher, “Defocused imaging exploits supercritical-angle fluorescence emission for precise axial single molecule localization microscopy,” Biomed. Opt. Express 11(2), 775–790 (2020). [CrossRef]
