A Simple and Efficient Method to Observe Internal Structures of Helminths by Scanning Electron Microscopy

Adnet, Fernando Antônio de O.; Gonçalves, Júlia P.; Souza, Wanderley de; Attias, Márcia

doi:10.1017/s1431927613013706

Cited by 3 publications

(4 citation statements)

References 16 publications

(28 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Over the last decade, the cryofracture technique has been proved a simple method used to prepare biological samples by exposing internal structures for visualization using SEM (Taylor, 2008). Indeed, previous studies have provided valuable new information about parasitic flukes (Adnet et al ., 2013). As mentioned above, in order to better characterize the uterine eggs observed by CM, a detailed examination using the cryofracture technique was performed.…”

Section: Discussionmentioning

confidence: 99%

“…Ten specimens were processed for SEM based on a previous protocol (Lopes-Torres et al ., 2013) and some specimens were frozen and fractured following a simple methodology previously described (Adnet et al ., 2013). Briefly, samples were dehydrated in a graded ethanol series (70–100% Degree Gay Lussac) for 40 minutes at each step, critical-point dried in CO 2 , mounted on metallic stubs and coated with gold (20–25 nm deposited).…”

Section: Methodsmentioning

confidence: 99%

“…The ultrastructural organization of E. coelomaticum cercariae, mother and daughter sporocysts (Franco-Acuña et al ., 2011; Pinheiro et al ., 2011, 2012), and eggs (Pinheiro et al ., 2015) has been investigated using scanning and transmission electron microscopies. Other works have already used advanced microscopy experiments for investigating nematodes (Adnet et al ., 2013; Lopes-Torres et al ., 2015), cestodes (Almeida et al ., 2015) and some trematode species (Machado-Silva et al ., 1998; Souza et al ., 2011, 2017; Gonçalves et al ., 2016; Marques et al ., 2017). Despite the excellent resolution to characterize the surface structures in detail, conventional scanning electron microscopies (SEMs) limit the observation to the external surfaces, missing information about the internal ones.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Eurytrema coelomaticum: updated morphology of adult worms using advanced microscopy experiments

et al. 2020

View full text Add to dashboard Cite

Eurytrema coelomaticum is a digenean flatworm of ruminants that is the causative agent of eurytrematosis, a disease of veterinary health concern. Although modern techniques of morphological analysis have provided new insights about the morphology and anatomy of parasitic helminths, most studies on E. coelomaticum adults are based on conventional light microscopy. In the present study, a combined approach using brightfield, fluorescence, confocal and scanning electron microscopies (SEMs), together with the cryofracture technique, have updated morphological data on E. coelomaticum recovered from cattle in Rio de Janeiro State, Brazil. Light microscopy confirmed the presence of several structures present in the current description, such as suckers, pharynx, oesophagus, intestinal bifurcation and the cirrus-sac. Fluorescence stereomicroscopy revealed for the first time the cubic crystal protein inclusions in the forebody, which were further detailed by confocal and SEMs. Confocal microscopy provided detailed information of the muscular architecture associated with the attachment structures (suckers), digestive system (pharynx and oesophagus), egg-forming complex (ovary, Mehlis’ gland and Laurer's canal) and male reproductive system, which are similar to those found in other digenean flukes. SEM images of cryofractured parasites showed mucus and developing eggs within uterine loops. It was demonstrated that the combination of advanced tools generated complementary information, confirming the importance of experimental morphology in parasitology. Therefore, the knowledge of the adult structural organization of E. coelomaticum was improved and this work has contributed to propose new morphological criteria to evaluate the effects of antiparasitic drugs on flukes of medical and veterinary importance.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Eurytrema coelomaticum: updated morphology of adult worms using advanced microscopy experiments

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Additional samples of barbs of the same feather and further sectioning of the same material above were observed in Scanning Electron Microscopy (SEM) and in Transmission Electron Microscopy (TEM) to verify some of the findings using STEM. TEM (Jeol 1200 EX, at HV = 80 kV) images were used to understand the internal characteristics of regular melanosomes and their overall shape, which were then corroborated by cryofractured barbs (preparation following Adnet et al, 2013) observed in a SEM microscope (Phillips XL 30, at HV = 12 kV and WD = 8 mm) (Supplementary Material online). The surface features found in the STEM images (see below), were verified on further sections observed in a SEM Zeiss Auriga (FIB-SEM) at HV = 0.5-3.0 kV and WD = 4-8 mm (Supplementary Material online).…”

Section: Sample Preparation and Observationmentioning

confidence: 99%

Quantitative characterization and comparative study of feather melanosome internal morphology using surface analysis

Straker

2016

Micron

View full text Add to dashboard Cite

A successful feather development implies in a precise orchestration of cells in the follicle, which culminates in one of the most complex epidermal structures in nature. Melanocytes contribute to the final structure by delivering melanosomes to the barb and barbule cells. Disturbance to the tissue during the feather growth can damage the final structure. Here, melanosomes seen in an unusual outgrowth on the barb cortex of a flight feather are reported and compared to commonly observed melanosomes embedded in the cortex. Transmission Electron Microscopy in scanning-transmission mode (STEM) generated images coupled with secondary electron detection. The two classes of melanosomes were registered on images combining transmitted and secondary electron signals. Image processing allowed surface analyses of roughness and texture of the internal morphology of these organelles. Results showed that the two classes of melanosomes are significantly distinct internally, indicating that different physiological processes up to feather maturation could have occurred. Surface analysis methods are not regularly used in cell biology studies, but here it is shown that it has great potential for microscopic image analysis, which could add robust information to studies of cell biology events.

show abstract

Additional characterization of the adult worm Mammomonogamus laryngeus (Railliet, 1899) and the tissue lesions caused by the infection in buffaloes

Torres

Pinheiro

Rodrigues

et al. 2020

Veterinary Parasitology

View full text Add to dashboard Cite

A Simple and Efficient Method to Observe Internal Structures of Helminths by Scanning Electron Microscopy

Cited by 3 publications

References 16 publications

Eurytrema coelomaticum: updated morphology of adult worms using advanced microscopy experiments

Eurytrema coelomaticum: updated morphology of adult worms using advanced microscopy experiments

Quantitative characterization and comparative study of feather melanosome internal morphology using surface analysis

Additional characterization of the adult worm Mammomonogamus laryngeus (Railliet, 1899) and the tissue lesions caused by the infection in buffaloes

Contact Info

Product

Resources

About