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Distinguishing calliphorid puparia through application of confocal laser scanning microscopy
The Calliphoridae, or blow flies, are of much ecological and practical importance given their roles in decompositional ecology, medical and veterinary myiasis, and forensic entomology. As ephemeral and rapidly developing species, adults are frequently not present for identification, but puparia, the remaining outer integument of the third instar larvae, frequently are found. These heavily sclerotized remains are stable in the environment, but are character conservative. Historically, scanning electron microscopy (SEM) has been used for characterization, a technique which is not only time consuming but often expensive, effectively making large numbers of specimens impossible to quantify. As an alternative, confocal laser scanning laser microscopy (CSLM) was tested for utility in providing superior data over SEM. Furthermore, due the use of intrinsic autofluorescence for imagining, CSLM is significantly more rapid than SEM requiring no preparation for imaging. Three channels of excitation and emission spectra provided not only image data from the pupal wall, but also from the hydrocarbons found upon the puparia. The laser excitation wavelengths were: 404.7, 488, 640.5 nm and the emissions were: 425–475, 500–550, 663–738 nm. For ten species of Calliphorids, CSLM was used to image puparia. Not only did this provide characters for species identification, but also allowed for the examination of hundreds of specimens. The data generated from the maximum intensity projection images was then used for multivariate discriminate analysis to test for characters useful in delimiting taxa. This provides a basis for puparia identification regardless of image methodology. ^
Elowsky, Christian G, "Distinguishing calliphorid puparia through application of confocal laser scanning microscopy" (2016). ETD collection for University of Nebraska - Lincoln. AAI10102759.