Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software.
Photon event centroiding in photon counting imaging and single-molecule localisation in super-resolution fluorescence microscopy share many traits. Although photon event centroiding has traditionally been performed with simple single-iteration algorithms, we recently reported that iterative fitting algorithms originally developed for single-molecule localisation fluorescence microscopy work very well when applied to centroiding photon events imaged with an MCP-intensified CMOS camera. Here, we have applied these algorithms for centroiding of photon events from an electron-bombarded CCD (EBCCD). We find that centroiding algorithms based on iterative fitting of the photon events yield excellent results and allow fitting of overlapping photon events, a feature not reported before and an important aspect to facilitate an increased count rate and shorter acquisition times.
Department of Physics, Kings College London, Strand, London WC2R 2LS, UK.
Liisa M
LM
Hirvonen
Department of Physics, Kings College London, Strand, London WC2R 2LS, UK.
Matthew J
MJ
Barber
Department of Physics, Kings College London, Strand, London WC2R 2LS, UK.
Klaus
K
Suhling
Electron-bombarded CCD; Single photon counting; Single-molecule localisation