Improving image quality of Rhodnius prolixus head using different types of staining methods and synchrotron radiation phase contrast microtomography

Sena, Gabriela; Nogueira, Liebert Parreiras; Braz, Delson; Colaço, Marcos V.; Almeida, André Pereira de; Gonzalez, Marcelo S.; Soares, Igor Masullo; Barroso, R.C.

doi:10.1016/j.radphyschem.2018.06.039

Cited by 5 publications

(4 citation statements)

References 14 publications

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“…For instance, a study has optimized the use of PTA for imaging of bumblebee brain structures, whereby PTA outperformed iodine and other staining solution in terms of resolution of microstructures, although iodine was faster to perfuse through the samples (Smith et al 2016). Comparing the advantage and disadvantages of different protocols present in the specialized literature, iodine makes it more suitable for studying whole insect specimens (Metscher 2009b;Du Plessis et al 2017;Sena et al 2019). Thus, it is important to balance the pros and cons of each staining method according to the type of sample, costs, time, and the resolution needed for the images.…”

Section: Discussionmentioning

confidence: 99%

Assessing Anatomical Changes in Male Reproductive Organs in Response to Larval Crowding Using Micro-computed Tomography Imaging

et al. 2022

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Ecological conditions shape (adaptive) responses at the molecular, anatomical, and behavioral levels. Understanding these responses is key to predict the outcomes of intra- and inter-specific competitions and the evolutionary trajectory of populations. Recent technological advances have enabled large-scale molecular (e.g., RNAseq) and behavioral (e.g., computer vision) studies, but the study of anatomical responses to ecological conditions has lagged behind. Here, we highlight the role of X-ray micro-computed tomography (micro-CT) in generating in vivo and ex vivo 3D imaging of anatomical structures, which can enable insights into adaptive anatomical responses to ecological environments. To demonstrate the application of this method, we manipulated the larval density of Drosophila melanogaster Meigen flies and applied micro-CT to investigate the anatomical responses of the male reproductive organs to varying intraspecific competition levels during development. Our data is suggestive of two classes of anatomical responses which broadly agree with sexual selection theory: increasing larval density led to testes and ejaculatory duct to be overall larger (in volume), while the volume of accessory glands and, to a lesser extent, ejaculatory duct decreased. These two distinct classes of anatomical responses might reflect shared developmental regulation of the structures of the male reproductive system. Overall, we show that micro-CT can be an important tool to advance the study of anatomical (adaptive) responses to ecological environments.

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Section: Discussionmentioning

confidence: 99%

Assessing Anatomical Changes in Male Reproductive Organs in Response to Larval Crowding Using Micro-computed Tomography Imaging

et al. 2022

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“…For instance, a study has optimised the use of PTA for imaging of bumblebee brain structures, whereby PTA outperformed iodine and other staining solution in terms of resolution of micro-structures, although iodine was faster to perfuse through the samples (Smith et al 2016). Comparing the advantage and disadvantages of different protocols present in the specialized literature, iodine makes it more suitable for studying whole insect specimens (Metscher 2009b; Du Plessis et al 2017; Sena et al 2019). Thus, it is important to balance the pros and cons of each staining method according to the type of sample, costs, time, and the resolution needed for the images.…”

Section: Discussionmentioning

confidence: 99%

Assessing anatomical changes in male reproductive organs in response to larval crowding using micro-computed tomography imaging

Morimoto

Barcellos

Schoborg

et al. 2021

Preprint

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Selection favour males that anticipate intraspecific competition levels and develop appropriate competitive traits. In holometabolous insects, larval density can act as an ecological cue of males future intraspecific competition levels while also limiting access to food allocated to sexually selected traits (e.g., testes). To date, however, most studies have been conducted in larval densities that may have little or no relevance to species ecology. Here, we manipulated Drosophila melanogaster larval density based on the natural history of the species and used 3D micro-CT imaging to show a strong modulation of testes volume based on larval density: males from high larval densities generally had larger testes. Conversely, males from high densities tended (albeit not statistically significantly) to have the smallest accessory glands and ejaculatory bulb. Overall, we used micro-CT to measure male reproductive morphology in an ecologically relevant design for which the findings broadly support predictions from sexual selection theory.

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“…To improve results with synchrotron microtomography and a polychromatic beam, after 2018 it was developed a staining protocol described by Sena et al. that allowed a better visualisation and segmentation of the Rhodnius prolixus structures 15,16 …”

Section: Introductionmentioning

confidence: 99%

“…14 To improve results with synchrotron microtomography and a polychromatic beam, after 2018 it was developed a staining protocol described by Sena et al that allowed a better visualisation and segmentation of the Rhodnius prolixus structures. 15,16 Some studies using laboratory microtomography conventional scanners combining with the correct staining methods have proved to be a powerful tool in biological research, allowing the visualisation of low-density tissues. [17][18][19][20] The main goal of the present work was to use staining protocols to study Rhodnius prolixus head anatomy with laboratory microtomography conventional scanners.…”

Section: Introductionmentioning

confidence: 99%

Investigation of Rhodnius prolixus head anatomy using microtomography

Sena

Silva

Nogueira

et al. 2023

Journal of Microscopy

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In the last years, microtomography has proved to be a powerful technique on insects' studies, allowing a detailed view of the structures' internal with a high resolution. One of the most important advantages about the use of microtomography in these studies is the fact that the dissection is not necessary, which decreases considerably the number of samples used on the insects' research. Some insects are used constantly in studies about morphology, metamorphosis, and reproduction, because they work as a model for others, and Rhodnius prolixus is one of the most studied in this group. This insect is also one of the main insect vectors of Chagas disease that kills around 12,000 people every year in Latin America. Some studies using laboratory microtomography conventional scanners combining with the correct staining methods have proved that it could be a powerful tool in biological research, allowing the visualisation of low-density tissues. The main goal of the present work was to use staining protocols to study Rhodnius prolixus with laboratory microtomography conventional scanners. The experiments were carried out at the imaging lab in the Theoretical Biology Department, University of Vienna, using an Xradia MicroXCT and at the University of Oslo, using a Skyscan 2211.

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Improving image quality of Rhodnius prolixus head using different types of staining methods and synchrotron radiation phase contrast microtomography

Cited by 5 publications

References 14 publications

Assessing Anatomical Changes in Male Reproductive Organs in Response to Larval Crowding Using Micro-computed Tomography Imaging

Assessing Anatomical Changes in Male Reproductive Organs in Response to Larval Crowding Using Micro-computed Tomography Imaging

Assessing anatomical changes in male reproductive organs in response to larval crowding using micro-computed tomography imaging

Investigation of Rhodnius prolixus head anatomy using microtomography

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