Preparation of dipteran larvae for scanning electron microscopy using a freeze-substitution technique

1986 ◽  
Vol 64 (3) ◽  
pp. 797-799 ◽  
Author(s):  
Douglas D. Colwell ◽  
Eric G. Kokko
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Freeze Substitution ◽  
Rapid Freezing ◽  
Fixation Methods ◽  
Good Preservation ◽  
Larval Stages ◽  
Excellent Preservation ◽  
Standard Fixation ◽  
Scanning Electron

Difficulties are often encountered in achieving good preservation of dipteran larvae for scanning electron microscopy. Standard fixation methods frequently fail because fixatives do not penetrate the cuticle rapidly, resulting in distortion and generation of artifacts on the surface. A freeze-substitution technique that produces excellent preservation of the larval stages of a wide variety of Diptera is described. The method employs Freon 12 for rapid freezing of the specimens and methanol as the substitution fluid. The technique is simple, inexpensive, and demonstrates a significant improvement in preservation of specimens.

scholarly journals Morphological aspects of immature stages of Migonemyia migonei (Diptera: Psychodidae, Phlebotominae), an important vector of Leishmaniosis in South America, described by scanning electron microscopy

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0242163
Author(s):  
Eric Fabrício Marialva ◽  
Nágila F. Secundino ◽  
Fernando F. Fernandes ◽  
Helena R. C. Araújo ◽  
Claudia M. Ríos-Velásquez ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
South America ◽  
Leishmania Infantum ◽  
Immature Stages ◽  
Vector Species ◽  
Future Studies ◽  
Larval Stages ◽  
Morphological Aspects ◽  
Scanning Electron

We describe the immature stages of Migonemyia migonei, which is the vector of Leishmania (Viannia) braziliensis, the etiological agent of cutaneous leishmaniasis in South America, and a putative vector of Leishmania infantum chagasi. Scanning Electron Microscopy (SEM) was used to refine the description of the structures of the egg, all instar larvae, and the pupa. The eggs have polygonal cells on the egg exochorion, and differences between larval and pupal chaetotaxy have been highlighted. Different sensillary subtypes—trichoidea, basiconica, coelonica and campanoformia—were observed in the larval stages. The results presented herein contribute to the taxonomy of Mg. migonei and may contribute to future studies on the phylogeny of this important vector species.

Description of the larval stages ofAnguillicola crassus(Nematoda, Dracunculoidea) using light and scanning electron microscopy

1992 ◽  
Vol 5 (4) ◽  
pp. 307-318 ◽  
Author(s):  
Guillailme Blanc ◽  
Stéphane Bonneau ◽  
Sylvie Biagianti ◽  
Annie J. Petter
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Larval Stages ◽  
Scanning Electron

A COMPARISON OF THE SENSORY MORPHOLOGY OF THE CEPHALIC REGION OF FIRST- AND THIRD-INSTAR ARACHNIDOMYIA ALDRICHI (PARKER) (DIPTERA: SARCOPHAGIDAE)

1993 ◽  
Vol 125 (1) ◽  
pp. 101-111 ◽  
Author(s):  
J.M. Schmidt
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Freeze Substitution ◽  
Cephalic Region ◽  
The Other ◽  
Sensory Structures ◽  
Scanning Electron

AbstractThe sensory structures of the cephalic segment of first- and third-instar Arachnidomyia aldrichi (Parker) (Diptera: Sarcophagidae) larvae were examined by scanning electron microscopy. Larvae were prepared by methanol freeze substitution, which effectively preserved the structure of these membranous specimens. Sensilla of first and third instars are described and compared with respect to differences in the sensory requirements of these instars. Sensilla numbers and types were similar in both instars, although the sensilla of third instars were larger than those of first instars. None of the external sensory structures was unique to one or the other instar. The cephalic sensory structures of A. aldrichi are also similar in arrangement and structure to those reported for other saprophagous and herbivorous muscomorph dipteran larvae.

scholarly journals Ossicle development of the crinoid Florometra serratissima through larval stages

2017 ◽  
Vol 95 (3) ◽  
pp. 183-192 ◽  
Author(s):  
Ariane Comeau ◽  
Cory D. Bishop ◽  
Christopher B. Cameron
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Soft Tissues ◽  
Sister Group ◽  
Coralline Algae ◽  
Rapid Radiation ◽  
Larval Stages ◽  
Evolution And Development ◽  
Feather Star ◽  
Scanning Electron

Crinoids are the oldest living class of echinoderm and sister group to the remaining eleutherozoan clade and so are key to discussions on the evolution and development of the echinoderm skeleton. Here we present the intraspecific variation of ossicle development of the feather star Florometra serratissima (A.H. Clark, 1907) during its three larval stages: doliolaria, cystidean, and early pentacrinoid. To induce settlement, larvae were cultured on a sea table in glass bowls containing coralline algae. The soft tissues of 60 larvae were dissolved to isolate and to observe the ossicles with compound microscopy and scanning electron microscopy. From the late doliolaria stage to 56-day-old pentacrinoids, a total of four types of ossicle developed: oral plates, basal plates, columnar stalk ossicles, and an attachment disk. Occasionally, an additional plate was found under the basal plates, which may represent a vestigial infrabasal plate. The shape of the attachment disk was plastic to accommodate the substrate. Crinoid ossicle development is variable in size, shape, and number, and the timing of development is asynchronous; traits that may have contributed to the early rapid radiation and phenotypic disparity of echinoderms.

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