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Anti-Stokes fluorescence from endogenously formed protoporphyrin IX - Implications for clinical multiphoton diagnostics.

Journal article
Authors Despoina Kantere
Stina Guldbrand
John Paoli
Mattias Goksör
Dag Hanstorp
Ann-Marie Wennberg
Maria Smedh
Marica B Ericson
Published in Journal of biophotonics
Volume 6
Issue 5
Pages 409–415
ISSN 1864-0648
Publication year 2013
Published at Institute of Clinical Sciences, Department of Dermatology and Venereology
Department of Physics (GU)
Core Facilities, Centre for Cellular Imaging
Pages 409–415
Language en
Subject categories Dermatology and Venereal Diseases


Multiphoton imaging based on two-photon excitation is making its way into the clinics, particularly for skin cancer diagnostics. It has been suggested that endogenously formed protoporphyrin IX (PpIX) induced by aminolevulinic acid or methylaminolevulinate can be applied to improve tumor contrast, in connection to imaging of tissue autofluorescence. However, previous reports are limited to cell studies and data from tissue are scarce. No report shows conclusive evidence that endogenously formed PpIX increases tumor contrast when performing multiphoton imaging in the clinical situation. We here demonstrate by spectral analysis that two-photon excitation of endogenously formed PpIX does not provide additional contrast in superficial basal cell carcinomas. In fact, the PpIX signal is overshadowed by the autofluorescent background. The results show that PpIX should be excited at a wavelength giving rise to one-photon anti-Stokes fluorescence, to overcome the autofluorescent background. Thus, this study reports on a plausible method, which can be implemented for clinical investigations on endogenously formed PpIX using multiphoton microscopy (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

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