Confocal laser scanning microscopy as a valuable tool in Diptera larval morphology studies
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Confocal laser scanning microscopy ...
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Issue date
2014-09-19Submitted date
2020-09-07Subject Terms
DipteraCyclorrhapha
Larva
Morphology
Confocal laser scanning microscope
Clearing technique
Metadata
Show full item recordAbstract
Larval morphology of flies is traditionally studied using light microscopy, yet in the case of fine structures compound light microscopy is limited due to problems of resolution, illumination and depth of field, not allowing for precise recognition of sclerites’ edges and interactions. Using larval instars of cyclorrhaphan Diptera, we show the usefulness of confocal laser scanning microscopy (CLSM) for studying the morphological characters of immature stages by taking advantage of the autofluorescent properties of cephaloskeleton structures. We compare data obtained from killed but unprepared larvae with those from larvae prepared by clearing according to two commonly used methods, either with potassium hydroxide or with Hoyer’s medium. We also evaluated the CLSM application for examining already slide-mounted larvae stored in museum collections and those freshly prepared. Our results indicate that CLSM and 3D reconstruction are excellent for visualizing small, compound structures of cylrorrhaphan larvae cephaloskeleton, if appropriate clearing techniques, i.e. the application of KOH, are used. Maximum intensity projection of confocal data sets obtained from material freshly prepared and that stored in museum collection does not differ. Because of this and the fact that KOH is commonly used as a clearing method to examine the cephaloskeleton of Diptera larvae, it is possible, and highly recommended, to use slides already prepared with this method for re-examination by CLSM. We conclude that CLSM application can be an invaluable source of data for studies of larval morphology of Cyclorrhapha by way of taxonomic diagnoses, character identification and improvement in characters homologization.Citation
Grzywacz, A., Góral, T., Szpila, K. et al. Confocal laser scanning microscopy as a valuable tool in Diptera larval morphology studies. Parasitol Res 113, 4297–4302 (2014). https://doi.org/10.1007/s00436-014-4125-0Publisher
Springer Science and Business Media LLCJournal
Parasitology ResearchType
Journal ArticleItem Description
This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.NHM Repository
NHM Repository
ISSN
0932-0113EISSN
1432-1955ae974a485f413a2113503eed53cd6c53
10.1007/s00436-014-4125-0
Scopus Count