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Clearing pigmented insect cuticle to investigate small insects' organs in situ using confocal laser-scanning microscopy (CLSM)

Clearing pigmented insect cuticle to investigate small insects' organs in situ using confocal laser-scanning microscopy (CLSM)

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Prüfsumme: MD5:bb0d0f978a934b1b73534dcc4676f506

SMOLLA, Marco, Markus RUCHTY, Manuel NAGEL, Christoph J. KLEINEIDAM, 2014. Clearing pigmented insect cuticle to investigate small insects' organs in situ using confocal laser-scanning microscopy (CLSM). In: Arthropod Structure & Development. 43(2), pp. 175-181. ISSN 1467-8039. eISSN 1873-5495. Available under: doi: 10.1016/j.asd.2013.12.006

@article{Smolla2014-03Clear-27890, title={Clearing pigmented insect cuticle to investigate small insects' organs in situ using confocal laser-scanning microscopy (CLSM)}, year={2014}, doi={10.1016/j.asd.2013.12.006}, number={2}, volume={43}, issn={1467-8039}, journal={Arthropod Structure & Development}, pages={175--181}, author={Smolla, Marco and Ruchty, Markus and Nagel, Manuel and Kleineidam, Christoph J.} }

2014-05-21T10:10:00Z Ruchty, Markus Nagel, Manuel Various microscopic techniques allow investigating structures from submicron to millimeter range, however, this is only possible if the structures of interest are not covered by pigmented cuticle. Here, we present a protocol that combines clearing of pigmented cuticle while preserving both, hard and soft tissues. The resulting transparent cuticle allows confocal laser-scanning microscopy (CLSM), which yields high-resolution images of e.g. the brain, glands, muscles and fine cuticular structures. Using a fluorescent dye, even single labeled neurons can be visualized and resolved up to an imaging depth of 150 μm through the cleared cuticle. Hydrogen-peroxide, which was used to clear the cuticle, does not preclude immunocytochemical techniques, shown by successful labeling of serotonin-immunoreactive neurons (5HT-ir) in the ants' brain. The ‘transparent insect protocol’ presented here is especially suited for small arthropods where dissection of organs is very demanding and difficult to achieve. Furthermore, the insect organs are preserved in situ thus allowing a more precise three-dimensional reconstruction of the structures of interest compared to, e.g., dissected or sectioned tissue. Nagel, Manuel Arthropod Structure & Development ; 43 (2014), 2. - S. 175-181 2014-03 eng Kleineidam, Christoph J. Smolla, Marco Ruchty, Markus deposit-license 2014-05-21T10:10:00Z Smolla, Marco Kleineidam, Christoph J. Clearing pigmented insect cuticle to investigate small insects' organs in situ using confocal laser-scanning microscopy (CLSM)

Dateiabrufe seit 01.10.2014 (Informationen über die Zugriffsstatistik)

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