King's College London

Research portal

Picosecond wide-field time-correlated single photon counting fluorescence microscopy with a delay line anode detector

Research output: Contribution to journalArticle

Liisa Maija Hirvonen, Wolfgang Becker, James Milnes, Thomas Conneely, Stefan Smietana, Alix Marie Le Marois, Ottmar Jagutzki, Klaus Suhling

Original languageEnglish
Article number071101
Number of pages4
JournalAPPLIED PHYSICS LETTERS
Volume109
Issue number7
DOIs
Accepted/In press3 Aug 2016
Published15 Aug 2016

Documents

  • Picosecond wide-field time_HIRVONEN_Accepted 3Aug2016_GOLD VoR

    Picosecond_wide_field_time_HIRVONEN_Accepted3August2016_GOLD_VoR.pdf, 918 KB, application/pdf

    Uploaded date:13 Sep 2016

    Version:Final published version

    Licence:CC BY

    © 2016 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

King's Authors

Abstract

We perform wide-field time-correlated single photon counting-based fluorescence lifetime imaging (FLIM) with a crossed delay line anode image intensifier, where the pulse propagation time yields the photon position. This microchannel plate-based detector was read out with conventional fast timing electronics and mounted on a fluorescence microscope with total internal reflection (TIR) illumination. The picosecond time resolution of this detection system combines low illumination intensity of microwatts with wide-field data collection. This is ideal for fluorescence lifetime imaging of cell membranes using TIR. We show that fluorescence lifetime images of living HeLa cells stained with membrane dye di-4-ANEPPDHQ exhibit a reduced lifetime near the coverslip in TIR compared to epifluorescence FLIM.

Download statistics

No data available

View graph of relations

© 2020 King's College London | Strand | London WC2R 2LS | England | United Kingdom | Tel +44 (0)20 7836 5454