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Proceedings Paper

Two-photon scanning systems for clinical high resolution in vivo tissue imaging
Author(s): K. König; J. Müller; M. Höfer; C. Müller; M. Weinigel; R. Bückle; P. Elsner; M. Kaatz; B. Messerschmidt
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Paper Abstract

The femtosecond laser multiphoton tomograph DermaInspect as well as high NA two-photon GRIN microendoscopes for in vivo tomography of human skin have been used to detect malignant melanoma as well as to study the diffusion and intradermal accumulation of topically applied cosmetical and pharmaceutical components. So far, more than 500 patients and volunteers in Europe, Australia, and Asia have been investigated with this unique tomograph. Near infrared 80 MHz picojoule femtosecond laser pulses were employed to excite endogenous fluorophores such as NAD(P)H, flavoproteins, melanin, and elastin as well as fluorescent components of a variety of ointments via a twophoton excitation process. In addition, collagen has been imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Individual intratissue cells, mitochondria, melanosomes, and the morphology of the nuclei as well as extracellular matrix elements could be clearly visualized due to molecular imaging and the calculation of fluorescence lifetime images. Nanoparticles and intratissue drugs have been detected non-invasively, in situ and over a period of up to 3 months. In addition, hydration effects and UV effects were studied by monitoring modifications of cellular morphology and autofluorescence. The system was used to observe the diffusion through the stratum corneum and the accumulation and release of functionalized nanoparticles along hair shafts and epidermal ridges. The DermaInspect been also employed to gain information on skin age and wound healing in patients with ulcers. Novel developments include a galvo/piezo-scan driven flexible articulated arm as well as a piezoscan-driven flexible high-NA GRIN microendoscope with photonic crystal fiber.

Paper Details

Date Published: 12 March 2008
PDF: 6 pages
Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 686014 (12 March 2008); doi: 10.1117/12.762986
Show Author Affiliations
K. König, JenLab GmbH (Germany)
Univ. of the Saarland (Germany)
J. Müller, JenLab GmbH (Germany)
M. Höfer, JenLab GmbH (Germany)
C. Müller, JenLab GmbH (Germany)
M. Weinigel, JenLab GmbH (Germany)
R. Bückle, JenLab GmbH (Germany)
P. Elsner, Univ. Jena (Germany)
M. Kaatz, Univ. Jena (Germany)
B. Messerschmidt, Grintech GmbH (Germany)

Published in SPIE Proceedings Vol. 6860:
Multiphoton Microscopy in the Biomedical Sciences VIII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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