Non-invasive imaging of Ixodes ricinus ticks using microCT

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F19%3AN0000157" target="_blank" >RIV/00027162:_____/19:N0000157 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Non-invasive imaging of Ixodes ricinus ticks using microCT

Popis výsledku v původním jazyce

Poster. Tick Ixodes ricinus is a common vector of Borrelia sp. Although mechanisms of bacteria distribution within ticks have been described, exact effects of Borrelia spp. on Ixodes internal structure is little known. Internal and external structure of ticks is commonly visualized using light, electron microscopy or histology techniques. Recently, computed X-ray tomography (microCT) has been used as a novel tool for investigation of morphologic features in insect. In comparison to electron microscopy and histology, microCT enables fast and easy sample processing, examination of several specimen at the same time. While final resolution of images from microCT scans is sufficient (2.5 to 6 µm), major challenge of X-ray scanning consists in low contrast of internal structures. Aim of the study was to develop and optimize protocol for microCT scanning of both unfed and fully fed ticks of I. ricinus. Unfed ticks were dehydrated in a graded 70-100% ethanol, then incubated in 2% osmium tetraoxide and dried using hexamethyldisilazane. Another group of unfed ticks were dried by letting them placed at room temperature for several weeks. All ticks were scanned on Bruker Skyscan 1276 with no filter, resolution 4 µm and 4K binning. Scanned data were reconstructed using Instarecon, further modified and analyzed using CTan software. 3-D rendered volumes of ticks were visualized in CTvox. Contrast and quality of internal structure did not differ between naturally dried and ethanol dehydrated ticks contrasted with osmium. In group of fed ticks, artificial in vitro blood feeding of the ticks with addition metal-based contrast agents will be carried out as next step. In conclusion, our study shows that microCT is a novel efficient tool for imaging of tick internal structure.

Název v anglickém jazyce

Non-invasive imaging of Ixodes ricinus ticks using microCT

Popis výsledku anglicky

Poster. Tick Ixodes ricinus is a common vector of Borrelia sp. Although mechanisms of bacteria distribution within ticks have been described, exact effects of Borrelia spp. on Ixodes internal structure is little known. Internal and external structure of ticks is commonly visualized using light, electron microscopy or histology techniques. Recently, computed X-ray tomography (microCT) has been used as a novel tool for investigation of morphologic features in insect. In comparison to electron microscopy and histology, microCT enables fast and easy sample processing, examination of several specimen at the same time. While final resolution of images from microCT scans is sufficient (2.5 to 6 µm), major challenge of X-ray scanning consists in low contrast of internal structures. Aim of the study was to develop and optimize protocol for microCT scanning of both unfed and fully fed ticks of I. ricinus. Unfed ticks were dehydrated in a graded 70-100% ethanol, then incubated in 2% osmium tetraoxide and dried using hexamethyldisilazane. Another group of unfed ticks were dried by letting them placed at room temperature for several weeks. All ticks were scanned on Bruker Skyscan 1276 with no filter, resolution 4 µm and 4K binning. Scanned data were reconstructed using Instarecon, further modified and analyzed using CTan software. 3-D rendered volumes of ticks were visualized in CTvox. Contrast and quality of internal structure did not differ between naturally dried and ethanol dehydrated ticks contrasted with osmium. In group of fed ticks, artificial in vitro blood feeding of the ticks with addition metal-based contrast agents will be carried out as next step. In conclusion, our study shows that microCT is a novel efficient tool for imaging of tick internal structure.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/EF15_003%2F0000495" target="_blank" >EF15_003/0000495: FIT (Farmakologie, Imunoterapie, nanoToxikologie)</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů