Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software

Liisa Maija Hirvonen; Matthew Barber; Klaus Suhling

doi:10.1016/j.nima.2016.03.024

Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software

Liisa Maija Hirvonen, Matthew Barber, Klaus Suhling

King's College London

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

217 Downloads (Pure)

Abstract

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.

Original language	English
Pages (from-to)	121-125
Journal	Nuclear Instruments & Methods In Physics Research Section A, Accelerators, Spectrometers, Detectors And Associated Equipment
Volume	820
Early online date	10 Mar 2016
DOIs	https://doi.org/10.1016/j.nima.2016.03.024
Publication status	Published - 1 Jun 2016

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2016_Hirvonen_NimAFinal published version, 1.61 MBLicence: CC BY

Cite this

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title = "Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software",

abstract = "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.",

author = "Hirvonen, {Liisa Maija} and Matthew Barber and Klaus Suhling",

year = "2016",

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doi = "10.1016/j.nima.2016.03.024",

language = "English",

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journal = "Nuclear Instruments & Methods In Physics Research Section A, Accelerators, Spectrometers, Detectors And Associated Equipment",

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Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software. / Hirvonen, Liisa Maija; Barber, Matthew; Suhling, Klaus.
In: Nuclear Instruments & Methods In Physics Research Section A, Accelerators, Spectrometers, Detectors And Associated Equipment, Vol. 820, 01.06.2016, p. 121-125.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software

AU - Hirvonen, Liisa Maija

AU - Barber, Matthew

AU - Suhling, Klaus

PY - 2016/6/1

Y1 - 2016/6/1

N2 - 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.

AB - 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.

U2 - 10.1016/j.nima.2016.03.024

DO - 10.1016/j.nima.2016.03.024

M3 - Article

SN - 0168-9002

VL - 820

SP - 121

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JO - Nuclear Instruments & Methods In Physics Research Section A, Accelerators, Spectrometers, Detectors And Associated Equipment

JF - Nuclear Instruments & Methods In Physics Research Section A, Accelerators, Spectrometers, Detectors And Associated Equipment

ER -

Photon counting imaging and centroiding with an electron-bombarded CCD using single molecule localisation software

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