Smart-aggregation imaging for single molecule localisation with SPAD cameras

Istvan Gyongy, Amy Davies, Neale A. W. Dutton, Rory R. Duncan, Colin Rickman, Robert K. Henderson, Paul A. Dalgarno

Research output: Contribution to journalArticle

Abstract

Single molecule localisation microscopy (SMLM) has become an essential part of the super-resolution toolbox for probing cellular structure and function. The rapid evolution of these techniques has outstripped detector development and faster, more sensitive cameras are required to further improve localisation certainty. Single-photon avalanche photodiode (SPAD) array cameras offer single-photon sensitivity, very high frame rates and zero readout noise, making them a potentially ideal detector for ultra-fast imaging and SMLM experiments. However, performance traditionally falls behind that of emCCD and sCMOS devices due to lower photon detection efficiency. Here we demonstrate, both experimentally and through simulations, that the sensitivity of a binary SPAD camera in SMLM experiments can be improved significantly by aggregating only frames containing signal, and that this leads to smaller datasets and competitive performance with that of existing detectors. The simulations also indicate that with predicted future advances in SPAD camera technology, SPAD devices will outperform existing scientific cameras when capturing fast temporal dynamics.

LanguageEnglish
Article number37349
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 23 Nov 2016

Fingerprint

Avalanches
Photons
Equipment and Supplies
Cellular Structures
Noise
Single Molecule Imaging
Technology

ASJC Scopus subject areas

  • General

Cite this

@article{0b92940b29144e48b98817f448e2d8cd,
title = "Smart-aggregation imaging for single molecule localisation with SPAD cameras",
abstract = "Single molecule localisation microscopy (SMLM) has become an essential part of the super-resolution toolbox for probing cellular structure and function. The rapid evolution of these techniques has outstripped detector development and faster, more sensitive cameras are required to further improve localisation certainty. Single-photon avalanche photodiode (SPAD) array cameras offer single-photon sensitivity, very high frame rates and zero readout noise, making them a potentially ideal detector for ultra-fast imaging and SMLM experiments. However, performance traditionally falls behind that of emCCD and sCMOS devices due to lower photon detection efficiency. Here we demonstrate, both experimentally and through simulations, that the sensitivity of a binary SPAD camera in SMLM experiments can be improved significantly by aggregating only frames containing signal, and that this leads to smaller datasets and competitive performance with that of existing detectors. The simulations also indicate that with predicted future advances in SPAD camera technology, SPAD devices will outperform existing scientific cameras when capturing fast temporal dynamics.",
author = "Istvan Gyongy and Amy Davies and Dutton, {Neale A. W.} and Duncan, {Rory R.} and Colin Rickman and Henderson, {Robert K.} and Dalgarno, {Paul A.}",
year = "2016",
month = "11",
day = "23",
doi = "10.1038/srep37349",
language = "English",
volume = "6",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

Smart-aggregation imaging for single molecule localisation with SPAD cameras. / Gyongy, Istvan; Davies, Amy; Dutton, Neale A. W.; Duncan, Rory R.; Rickman, Colin; Henderson, Robert K.; Dalgarno, Paul A.

In: Scientific Reports, Vol. 6, 37349, 23.11.2016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Smart-aggregation imaging for single molecule localisation with SPAD cameras

AU - Gyongy, Istvan

AU - Davies, Amy

AU - Dutton, Neale A. W.

AU - Duncan, Rory R.

AU - Rickman, Colin

AU - Henderson, Robert K.

AU - Dalgarno, Paul A.

PY - 2016/11/23

Y1 - 2016/11/23

N2 - Single molecule localisation microscopy (SMLM) has become an essential part of the super-resolution toolbox for probing cellular structure and function. The rapid evolution of these techniques has outstripped detector development and faster, more sensitive cameras are required to further improve localisation certainty. Single-photon avalanche photodiode (SPAD) array cameras offer single-photon sensitivity, very high frame rates and zero readout noise, making them a potentially ideal detector for ultra-fast imaging and SMLM experiments. However, performance traditionally falls behind that of emCCD and sCMOS devices due to lower photon detection efficiency. Here we demonstrate, both experimentally and through simulations, that the sensitivity of a binary SPAD camera in SMLM experiments can be improved significantly by aggregating only frames containing signal, and that this leads to smaller datasets and competitive performance with that of existing detectors. The simulations also indicate that with predicted future advances in SPAD camera technology, SPAD devices will outperform existing scientific cameras when capturing fast temporal dynamics.

AB - Single molecule localisation microscopy (SMLM) has become an essential part of the super-resolution toolbox for probing cellular structure and function. The rapid evolution of these techniques has outstripped detector development and faster, more sensitive cameras are required to further improve localisation certainty. Single-photon avalanche photodiode (SPAD) array cameras offer single-photon sensitivity, very high frame rates and zero readout noise, making them a potentially ideal detector for ultra-fast imaging and SMLM experiments. However, performance traditionally falls behind that of emCCD and sCMOS devices due to lower photon detection efficiency. Here we demonstrate, both experimentally and through simulations, that the sensitivity of a binary SPAD camera in SMLM experiments can be improved significantly by aggregating only frames containing signal, and that this leads to smaller datasets and competitive performance with that of existing detectors. The simulations also indicate that with predicted future advances in SPAD camera technology, SPAD devices will outperform existing scientific cameras when capturing fast temporal dynamics.

UR - http://www.scopus.com/inward/record.url?scp=84997514574&partnerID=8YFLogxK

U2 - 10.1038/srep37349

DO - 10.1038/srep37349

M3 - Article

VL - 6

JO - Scientific Reports

T2 - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 37349

ER -