Microsecond wide-field TCSPC microscopy based on an ultra-fast CMOS camera

Liisa M. Hirvonen, Zdeněk Petrášek, Andrew Beeby, Klaus Suhling

Research output: Chapter in Book/Conference paperConference paper

4 Citations (Scopus)

Abstract

Ultra-fast frame rate CMOS cameras, combined with a photon counting image intensifier, can be used for microsecond resolution wide-field time-correlated single photon counting (TCSPC) microscopy. A sequence of frames is recorded after an excitation pulse, and the number and location of photons in each frame is determined. This process is repeated until enough photons are recorded for a photon arrival time histogram in the pixels of the image. This approach combines low, nanowatt excitation power with single-photon detection sensitivity and arrival timing in many pixels simultaneously, short acquisition times in the order of seconds and allows lifetime mapping with a time resolution of ∼1 microsecond. Moreover, we also show that the phosphor decay can be exploited to time the photon arrival well below the exposure time of the camera. This approach yields better time resolution and larger images than direct imaging of photon events. We show that both methods are ideal for lifetime imaging of transition metal compounds in living cells within a few seconds.

Original languageEnglish
Title of host publicationMultiphoton Microscopy in the Biomedical Sciences XV
EditorsKarsten Konig, Peter T. C. So, Ammasi Periasamy
Place of PublicationUSA
PublisherSPIE-INT SOC OPTICAL ENGINEERING
Volume9329
ISBN (Electronic)9781628414196
DOIs
Publication statusPublished - 1 Jan 2015
Externally publishedYes
EventMultiphoton Microscopy in the Biomedical Sciences XV - San Francisco, United States
Duration: 8 Feb 201510 Feb 2015

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume9329
ISSN (Print)1605-7422

Conference

ConferenceMultiphoton Microscopy in the Biomedical Sciences XV
CountryUnited States
CitySan Francisco
Period8/02/1510/02/15

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  • Cite this

    Hirvonen, L. M., Petrášek, Z., Beeby, A., & Suhling, K. (2015). Microsecond wide-field TCSPC microscopy based on an ultra-fast CMOS camera. In K. Konig, P. T. C. So, & A. Periasamy (Eds.), Multiphoton Microscopy in the Biomedical Sciences XV (Vol. 9329). [932939] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 9329). SPIE-INT SOC OPTICAL ENGINEERING. https://doi.org/10.1117/12.2178567