Depth-dependent autofluorescence photobleaching using 325, 473, 633, and 785 nm of porcine ear skin ex vivo

Johannes Schleusener; Jürgen Lademann; Maxim E. Darvin

doi:10.1117/1.JBO.22.9.091503

5 January 2017 Depth-dependent autofluorescence photobleaching using 325, 473, 633, and 785 nm of porcine ear skin ex vivo

Johannes Schleusener, Jürgen Lademann, Maxim E. Darvin

Author Affiliations +

Journal of Biomedical Optics, Vol. 22, Issue 9, 091503 (January 2017). https://doi.org/10.1117/1.JBO.22.9.091503

Abstract

Autofluorescence photobleaching describes the decrease of fluorescence intensity of endogenous fluorophores in biological tissue upon light irradiation. The origin of autofluorescence photobleaching is not fully understood. In the skin, the spatial distribution of various endogenous fluorophores varies within the skin layers. Most endogenous fluorophores are excited in the ultraviolet and short visible wavelength range, and only a few, such as porphyrins (red) and melanin (near-infrared), are excited at longer wavelengths. The excitation wavelength- and depth-dependent irradiation of skin will therefore excite different fluorophores, which will likely influence the photobleaching characteristics. The autofluorescence photobleaching of porcine ear skin has been measured ex vivo using 325, 473, 633, and 785 nm excitation at different skin depths from the surface to the dermis at 150 μm. Confocal Raman microscopes were used to achieve sufficient spatial resolution of the measurements. The autofluorescence area under the curve was measured for 21 consecutive acquisitions of 15 s. In all cases, the photobleaching follows a two-exponential decay function approximated by nonlinear regression. The results show that photobleaching can be applied to improve the signal-to-noise ratio in Raman spectroscopy for all of the applied excitation wavelengths and skin depths.

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Johannes Schleusener, Jürgen Lademann, and Maxim E. Darvin "Depth-dependent autofluorescence photobleaching using 325, 473, 633, and 785 nm of porcine ear skin ex vivo," Journal of Biomedical Optics 22(9), 091503 (5 January 2017). https://doi.org/10.1117/1.JBO.22.9.091503

Received: 9 September 2016; Accepted: 15 November 2016; Published: 5 January 2017

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KEYWORDS

Skin

Luminescence

Raman spectroscopy

In vivo imaging

Ultraviolet radiation

Ear

Confocal microscopy

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