Correcting Artifacts in Single Molecule Localization Microscopy Analysis Arising from Pixel Quantum Efficiency Differences in sCMOS Cameras
| Autor: | Hazen P. Babcock, Fang Huang, Colenso M. Speer |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Fluorescence-lifetime imaging microscopy Offset (computer science) lcsh:Medicine 02 engineering and technology Poisson distribution Article Fluorescence imaging Optical imaging 03 medical and health sciences symbols.namesake Mice Thalamus Microscopy Quantum Dots Image Processing Computer-Assisted Animals Poisson Distribution Super-resolution microscopy lcsh:Science Physics Multidisciplinary Pixel lcsh:R Equipment Design 021001 nanoscience & nanotechnology Single Molecule Imaging 030104 developmental biology Microscopy Fluorescence Semiconductors Calibration symbols Quantum efficiency lcsh:Q 0210 nano-technology Biological imaging Artifacts Algorithm Algorithms |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-10 (2019) |
| ISSN: | 2045-2322 |
| Popis: | Optimal analysis of single molecule localization microscopy (SMLM) data acquired with a scientific Complementary Metal-Oxide-Semiconductor (sCMOS) camera relies on statistical compensation for its pixel-dependent gain, offset and readout noise. In this work we show that it is also necessary to compensate for differences in the relative quantum efficiency (RQE) of each pixel. We found differences in RQE on the order of 4% in our tested sCMOS sensors. These differences were large enough to have a noticeable effect on analysis algorithm results, as seen both in simulations and biological imaging data. We discuss how the RQE differences manifest themselves in the analysis results and present the modifications to the Poisson maximum likelihood estimation (MLE) sCMOS analysis algorithm that are needed to correct for the RQE differences. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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