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United States Department of Agriculture

Agricultural Research Service

Research Project: IPM TECHNOLOGIES FOR INSECT PESTS OF ORCHARD CROPS

Location: Subtropical Insects and Horticulture Research

Title: From the sample preparation to the volume rendering images of small animals: A step by step example of a procedure to carry out the micro-CT study of the leafhopper insect Homalodisca vitripennis (Hemiptera: Cicadellidae)

Authors
item Alba-Tercedor, J -
item Hunter, Wayne

Submitted to: Meeting Proceedings
Publication Type: Proceedings
Publication Acceptance Date: March 28, 2014
Publication Date: May 5, 2014
Repository URL: http://www.skyscan.be/company/usersmeeting2014a.htm
Citation: Alba-Tercedor, J., Hunter, W.B. 2014. From the sample preparation to the volume rendering images of small animals: A step by step example of a procedure to carry out the micro-CT study of the leafhopper insect Homalodisca vitripennis (Hemiptera: Cicadellidae). In Proceeedings: Bruker Micro-CT Users Meeting 2014. p 260-288. Ed.BrukermicroCT.Kontich, Belgium (ISBN: 9789081678100, ISSN: 2033-8031).

Interpretive Summary: Examination of insect anatomy has changed due to advances in micro-CT, digital computed tomography (CT). Combining micro-CT with Digital Video Library systems, and linking this to Big Data, will change the way researchers, entomologist, and the public search and use anatomical information. Described are the methods used to scan and examine the anatomy of the glassy-winged sharpshooter leafhopper. This leafhopper spreads a plant infecting bacterium which cause Pierce’s disease of grapevines. The insect anatomy was elucidated using a high resolution Bruker Skyscan 1172 micro tomography system, micro-CT at the Department of Zoology, University of Granada, Spain. High resolution scanning combined with computer software enabled rotation, ‘digital sectioning’, and coloring of specimens providing an innovative, interactive system to explore specimen anatomy comparable to scanning electron microscopy.

Technical Abstract: Advances in micro-CT, digital computed tomography (CT) scan uses X-rays to make detailed pictures of structures inside of the body. Combining micro-CT with Digital Video Library systems, and linking this to Big Data, will change the way researchers, entomologist, and the public search and use anatomical information. Described are the methods used to scan and examine the anatomy of the glassy-winged sharpshooter leafhopper, Homalodisca vitripennis (Hemiptera: Cicadellidae), using this technology. This leafhopper spreads a plant infecting bacterium which cause Pierce’s disease of grapevines. The insect anatomy was elucidated using a high resolution Bruker Skyscan 1172 micro tomography system, micro-CT (www.Skyscan.be) at the Department of Zoology, University of Granada, Spain. Live leafhoppers were prepared by overnight fixation in 4% glutaraldehyde with 2.5% paraformaldehyde made with sodium cacodolate buffer pH 6.5. Samples were rinsed three times, 10 min each, with 30% ethanol, and dehydrated in an ethanol series, 30 min per step, 50%,70,80,90,95, three times at 100%. Samples were chemically dried by placing in 2 mL of 100% Hexamethyldisilazane (HDMDS) for 2 hours, drying overnight at 35C. High resolution scans combined with computer software enabled rotation, ‘digital sectioning’, and coloring of specimens providing an innovative, interactive system to explore specimen anatomy comparable to scanning electron microscopy, SEM. Video at https://www.youtube.com/watch?v=RcPeotDsBK8. An in-depth explanation of the sample preparations for leafhoppers and other insects plus the operation of the software for the Bruker SkyScan system is at: www.skyscan.be/company/usersmeeting2014a.

Last Modified: 8/21/2014
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