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Journal of Biomedical Optics • Open Access

Nonlinear microscopy of chitin and chitinous structures: a case study of two cave-dwelling insects
Author(s): Mihailo D. Rabasović; Dejan V. Pantelić; Branislav M. Jelenković; Srećko B. Ćurčić; Maja S. Rabasović; Maja D. Vrbica; Vladimir M. Lazović; Božidar P. M. Ćurčić; Aleksandar J. Krmpot

Paper Abstract

We performed a study of the nonlinear optical properties of chemically purified chitin and insect cuticle using two-photon excited autofluorescence (TPEF) and second-harmonic generation (SHG) microscopy. Excitation spectrum, fluorescence time, polarization sensitivity, and bleaching speed were measured. We have found that the maximum autofluorescence signal requires an excitation wavelength below 850 nm. At longer wavelengths, we were able to penetrate more than 150-μm deep into the sample through the chitinous structures. The excitation power was kept below 10 mW (at the sample) in order to diminish bleaching. The SHG from the purified chitin was confirmed by spectral- and time-resolved measurements. Two cave-dwelling, depigmented, insect species were analyzed and three-dimensional images of the cuticular structures were obtained.

Paper Details

Date Published: 9 January 2015
PDF: 10 pages
J. Biomed. Opt. 20(1) 016010 doi: 10.1117/1.JBO.20.1.016010
Published in: Journal of Biomedical Optics Volume 20, Issue 1
Show Author Affiliations
Mihailo D. Rabasović, Univ. of Belgrade (Serbia)
Dejan V. Pantelić, Univ. of Belgrade (Serbia)
Branislav M. Jelenković, Univ. of Belgrade (Serbia)
Srećko B. Ćurčić, Univ. of Belgrade (Serbia)
Maja S. Rabasović, Univ. of Belgrade (Serbia)
Maja D. Vrbica, Univ. of Belgrade (Serbia)
Vladimir M. Lazović, Univ. of Belgrade (Serbia)
Božidar P. M. Ćurčić, Univ. of Belgrade (Serbia)
Aleksandar J. Krmpot, Univ. of Belgrade (Serbia)

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