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Intravital multiphoton photoconversion with a cell membrane dye.

Raphaël Turcotte1, Juwell W Wu1, Charles P Lin1

  • 1Wellman Center for Photomedicine and Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, CPZN 8238, 185 Cambridge Street, Boston, MA 02114, USA.

Journal of Biophotonics
|July 20, 2016
PubMed
Summary

Di-8-ANEPPS, a membrane dye, exhibits multiphoton photoconversion under near-infrared laser pulses. This spectral shift from red to blue allows for long-term in vivo cell marking and tracking.

Keywords:
Photoconversionbone marrowcell trackingintravital microscopymultiphotontwo-photon fluorescence microscopy

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Area of Science:

  • Biophysics
  • Cell Biology
  • Optical Imaging

Background:

  • Photoconversion enables dynamic tracking of cellular structures.
  • Di-8-ANEPPS is a widely used membrane dye.

Purpose of the Study:

  • To investigate the photoconversion properties of Di-8-ANEPPS.
  • To assess its utility for in vivo cell tracking.

Main Methods:

  • Near-infrared femtosecond laser pulses were used for illumination.
  • Multiphoton photoconversion was analyzed by power dependence (ρpc∝Iexc2.27).
  • Spectral shifts were measured in vitro and in vivo.

Main Results:

  • Di-8-ANEPPS demonstrated multiphoton photoconversion with a supralinear power dependence.
  • The emission spectrum shifted from red to blue post-photoconversion.
  • A 65-fold increase in blue to red ratio was observed in vitro.
  • The spectral shift remained stable in vivo for hours.

Conclusions:

  • Di-8-ANEPPS is suitable for multiphoton photoconversion.
  • Its stable spectral shift facilitates long-term intravital cell marking and tracking.