Island Assay: En høy gjennomstrømningsmetode for å evaluere Drosophila Locomotor Behavior
Published: April 30, 2023
Abstract
Kilde: Eidhof, I., et al. High-throughput Analyse av lokomotorisk oppførsel i Drosophila Island Assay. J. Vis. Utt. (2017).
Denne videoen beskriver øyanalysen, en atferdsmetode som brukes til å studere Drosophila-bevegelse. Eksempelprotokollen har et oppsett for analysen som er kompatibel med halvautomatisk bildeanalyse.
Protocol
Denne protokollen er et utdrag fra Eidhof et al.,High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay, J. Vis. Exp. (2017). 1. Bygging av øya assay box Forbered et brett laget av et robust materiale, for eksempel polymetylmetakrylat (PMMA), for å inneholde et lag med vann (dvs. et bad). Pass på at materialet ikke er hvitt.MERK: Dimensjoner på 40 x 35 x 2,5 cm3 anbefales (<str…
Representative Results
Figur 1: Flytskjema som skisserer kravene, eksperimentell prosedyre og analyse av øyanalysen. (A) Analyseutstyr på øya. (B) Eksperimentelt oppsett for øyanalysen. (C) Øyanalyse. (D) Behandling av øyanalysedata med makroen”Drosophila Island Assay”. Makroen”Drosophila Island Assay” består av 3 undermakroer: 1) gjør stakk og projeks…
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).