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Optics Letters

Optics Letters


  • Editor: Xi-Cheng Zhang
  • Vol. 39, Iss. 15 — Aug. 1, 2014
  • pp: 4482–4485

Steady state anisotropy two-photon microscopy resolves multiple, spectrally similar fluorophores, enabling in vivo multilabel imaging

J. Matthew Dubach, Claudio Vinegoni, and Ralph Weissleder  »View Author Affiliations

Optics Letters, Vol. 39, Issue 15, pp. 4482-4485 (2014)

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The use of spectrally distinguishable fluorescent dyes enables imaging of multiple targets. However, in two-photon microscopy, the number of fluorescent labels with distinct emission spectra that can be effectively excited and resolved is constrained by the confined tuning range of the excitation laser and the broad and overlapping nature of fluorophore two-photon absorption spectra. This limitation effectively reduces the number of available imaging channels. Here, we demonstrate that two-photon steady state anisotropy imaging (2PSSA) offers the capability to resolve otherwise unresolvable fluorescent tracers both in live cells and in mouse tumor models. This approach expands the number of biological targets that can be imaged simultaneously, increasing the total amount of information that can be obtained through imaging.

© 2014 Optical Society of America

OCIS Codes
(110.0180) Imaging systems : Microscopy
(170.0110) Medical optics and biotechnology : Imaging systems
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:
Medical Optics and Biotechnology

Original Manuscript: May 5, 2014
Revised Manuscript: June 17, 2014
Manuscript Accepted: June 18, 2014
Published: July 25, 2014

Virtual Issues
Vol. 9, Iss. 10 Virtual Journal for Biomedical Optics

J. Matthew Dubach, Claudio Vinegoni, and Ralph Weissleder, "Steady state anisotropy two-photon microscopy resolves multiple, spectrally similar fluorophores, enabling in vivo multilabel imaging," Opt. Lett. 39, 4482-4485 (2014)

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