Island Assay: En high-throughput metode til at evaluere Drosophila Locomotor Behavior
Published: April 30, 2023
Abstract
Kilde: Eidhof, I., et al. High-throughput Analyse af lokomotorisk adfærd i Drosophila Island Assay. J. Vis. Exp. (2017).
Denne video beskriver øen assay, en adfærdsmæssig metode, der anvendes til at studere Drosophila bevægelse. Eksempelprotokollen indeholder en opsætning til analysen, der er kompatibel med halvautomatisk billedanalyse.
Protocol
Denne protokol er et uddrag fra Eidhof et al.,High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay, J. Vis. Exp. (2017). 1. Opførelse af øen Assay Box Forbered en bakke lavet af et robust materiale, såsom poly methyl methacrylat (PMMA), til at indeholde et lag vand(dvs. et bad). Sørg for, at materialet ikke er hvidt.BEMÆRK: Dimensioner på 40 x 35 x 2,5 cm3 anbefales<strong …
Representative Results
Figur 1: Rutediagram, der skitserer behovene, forsøgsproceduren og analysen af øens analyse. (A)Island assay udstyr. (B) Eksperimentel opsætning for øens analyse. (C)Island assay. (D) Behandling af ø-analysedata med makroen”Drosophila Island Assay”. Makroen”Drosophila Island Assay” består af 3 undermakroer: 1) lav stak og projektio…
Materials
| 25 x 95 mm Drosophila vials | Flystuff | 32-116SB | – |
| Logitech C525 HD Webcam | Logitech | – | Any webcam with USB connection is suitable. |
| Stand to hold webcam | – | – | – |
| Lamp | – | – | 12 V LED lights are appropriate |
| Pounding pad | – | – | Any mouse pad works |
| Island Assay box | – | – | Dimensions 40x35x2.5 cm. Hole 20×30 cm. Transparent. |
| Island Assay bath | – | – | Dimensions 42x38x25 cm. Non white. |
| Island/platform | – | – | Dimensions 42x38x25 cm. Uniform white. |
| Soap | – | – | Standard dishwashing detergent is suitable. |
| Computer | – | – | Scripts run both on Windows and Mac |
| Image-recording software: HandiAvi® | AZcendant® | – | HandyAvi is only compatible with Windows and has been described throughout the manuscript. It can be downloaded from: http://www.azcendant.com/DownloadHandyAvi.html (version 5.0) |
| Image-recording software: WebcamCapture | – | – | Fiji/ImageJ plugin that can be used on Mac alternative to HandyAvi for image-recordings and can be downloaded from: https://imagej.nih.gov/ij/plugins/webcam-capture/ When using this method, the user has to use the same folder setup and image-recording settings indicated in this manuscript, with the exception that for each experimental replicate, the captured image stack should be exported as Stack.tiff to the corresponding experimental replicate folder. Upon running the "Drosophila Island Assay" macro on this data, no text should be present in the "First frame identifier" setting. |
Cite This Article
Island Assay: A High-Throughput Method to Evaluate Drosophila Locomotor Behavior. J. Vis. Exp. (Pending Publication), e20103, doi: (2023).