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- W2018614124 abstract "EMCCDs are widely used in super-resolution and single molecule imaging methods. These techniques rely on photoelectron counts for particle detection and calculating localization precision of single molecules, making accurate calibration of EMCCD camera counts to photoelectrons unit necessary. Calibration methods used in literature include shot noise based calibration and manufacturer-provided formulas, but there is no general consensus and the accuracy of these methods has yet to be verified. First, we directly measure the light response of an EMCCD using a photon counting approach. We use a greatly attenuated laser to illuminate a few pixels on an EMCCD and compare the EMCCD counts to the readout of a photon counting photomultiplier tube placed in the same position. Second, we process uniformly illuminated EMCCD images to isolate shot noise as the dominant noise course to create an adjusted mean-variance method of calibrating EMCCDs. We verify this adjusted mean-variance method using our photon counting approach. Third, we compare these results to the theoretical values from the EMCCD calibration methods used in literature. To test our modified calibration method, we examine single molecule localization data and compared the data to existing models." @default.
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- W2018614124 date "2013-01-01" @default.
- W2018614124 modified "2023-10-16" @default.
- W2018614124 title "Accurate EMCCD Photoelectron Calibration for Single Molecule Imaging Techniques" @default.
- W2018614124 doi "https://doi.org/10.1016/j.bpj.2012.11.3678" @default.
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