OSA's Digital Library

Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Vol. 39, Iss. 34 — Dec. 1, 2000
  • pp: 6306–6311

Four-dimensional multiphoton microscopy with time-correlated single-photon counting

Andreas Schönle, Markus Glatz, and Stefan W. Hell  »View Author Affiliations


Applied Optics, Vol. 39, Issue 34, pp. 6306-6311 (2000)
http://dx.doi.org/10.1364/AO.39.006306


View Full Text Article

Enhanced HTML    Acrobat PDF (3153 KB)





Browse Journals / Lookup Meetings

Browse by Journal and Year


   


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools

Share
Citations

Abstract

We report on the implementation of fluorescence-lifetime imaging in multiphoton excitation microscopy that uses PC-compatible modules for time-correlated single-photon counting. Four-dimensional data stacks are produced with each pixel featuring fluorescence-decay curves that consist of as many as 4096 bins. Fluorescence lifetime(s) and their amplitude(s) are extracted by statistical methods at each pixel or in arbitrarily defined regions of interest. When employing an avalanche photodiode the width of the temporal response function is 420 ps. Although this response confines the temporal resolution to values greater than several hundreds of picoseconds, the lifetime precision is determined by the signal-to-noise ratio and can be in the range of tens of picosconds. Lifetime changes are visualized in pulsed-laser-deposited fluorescent layers as well as in cyan fluorescent proteins that transfer energy to yellow fluorescent proteins in live mammalian cells.

© 2000 Optical Society of America

OCIS Codes
(110.0110) Imaging systems : Imaging systems
(120.4640) Instrumentation, measurement, and metrology : Optical instruments
(180.2520) Microscopy : Fluorescence microscopy

History
Original Manuscript: July 5, 2000
Revised Manuscript: July 25, 2000
Published: December 1, 2000

Citation
Andreas Schönle, Markus Glatz, and Stefan W. Hell, "Four-dimensional multiphoton microscopy with time-correlated single-photon counting," Appl. Opt. 39, 6306-6311 (2000)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-39-34-6306

You do not have subscription access to this journal. Citation lists with outbound citation links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Log in to access OSA Member Subscription

You do not have subscription access to this journal. Cited by links are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Log in to access OSA Member Subscription

You do not have subscription access to this journal. Figure files are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Log in to access OSA Member Subscription

You do not have subscription access to this journal. Article level metrics are available to subscribers only. You may subscribe either as an OSA member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Log in to access OSA Member Subscription

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited