Research Papers: Imaging

Nonlinear microscopy of chitin and chitinous structures: a case study of two cave-dwelling insects

[+] Author Affiliations
Mihailo D. Rabasović

University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Zemun, Belgrade, Serbia

Dejan V. Pantelić

University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Zemun, Belgrade, Serbia

Branislav M. Jelenković

University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Zemun, Belgrade, Serbia

Srećko B. Ćurčić

University of Belgrade—Faculty of Biology, Institute of Zoology, Studentski Trg 16, 11000 Belgrade, Serbia

Maja S. Rabasović

University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Zemun, Belgrade, Serbia

Maja D. Vrbica

University of Belgrade—Faculty of Biology, Institute of Zoology, Studentski Trg 16, 11000 Belgrade, Serbia

Vladimir M. Lazović

University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Zemun, Belgrade, Serbia

Božidar P. M. Ćurčić

University of Belgrade—Faculty of Biology, Institute of Zoology, Studentski Trg 16, 11000 Belgrade, Serbia

Aleksandar J. Krmpot

University of Belgrade, Institute of Physics, Pregrevica 118, 11080 Zemun, Belgrade, Serbia

J. Biomed. Opt. 20(1), 016010 (Jan 09, 2015). doi:10.1117/1.JBO.20.1.016010
History: Received September 23, 2014; Accepted December 9, 2014
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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.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Mihailo D. Rabasović ; Dejan V. Pantelić ; Branislav M. Jelenković ; Srećko B. Ćurčić ; Maja S. Rabasović, et al.
"Nonlinear microscopy of chitin and chitinous structures: a case study of two cave-dwelling insects", J. Biomed. Opt. 20(1), 016010 (Jan 09, 2015). ; http://dx.doi.org/10.1117/1.JBO.20.1.016010


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