Print Email Facebook Twitter Photon count estimation in single-molecule localization microscopy Title Photon count estimation in single-molecule localization microscopy Author Thorsen, R.Ø. (TU Delft ImPhys/Quantitative Imaging) Hulleman, C.N. (TU Delft ImPhys/Quantitative Imaging) Mathias, Hammer (University of Massachusetts Medical School) Grunwald, David (University of Massachusetts Medical School) Stallinga, S. (TU Delft ImPhys/Imaging Physics) Rieger, B. (TU Delft ImPhys/Quantitative Imaging) Department ImPhys/Imaging Physics Date 2018 Abstract Recently, Franke, Sauer and van de Linde introduced a way to estimate the axial position of single-molecules (TRABI). To this end, they compared the detected photon count from a temporal radial-aperture-based intensity estimation to the estimated count from Gaussian point-spread function (PSF) fitting to the data. Empirically they found this photometric ratio to be around 0.7-0.8 close to focus and decreasing away from it. Here, we explain this reported but unexplained discrepancy and furthermore show that the photometric ratio as indicator for axial position is susceptible even to typical optical aberrations. To reference this document use: http://resolver.tudelft.nl/uuid:16f8cad4-6674-4679-82fc-b44bc490bf2b DOI https://doi.org/10.1101/396424 Part of collection Institutional Repository Document type working paper Rights © 2018 R.Ø. Thorsen, C.N. Hulleman, Hammer Mathias, David Grunwald, S. Stallinga, B. Rieger Files PDF 396424.full.pdf 2.34 MB Close viewer /islandora/object/uuid:16f8cad4-6674-4679-82fc-b44bc490bf2b/datastream/OBJ/view