FIM, a novel FTIR-based imaging method for high throughput locomotion analysis

PLoS One. 2013;8(1):e53963. doi: 10.1371/journal.pone.0053963. Epub 2013 Jan 21.

Abstract

We designed a novel imaging technique based on frustrated total internal reflection (FTIR) to obtain high resolution and high contrast movies. This FTIR-based Imaging Method (FIM) is suitable for a wide range of biological applications and a wide range of organisms. It operates at all wavelengths permitting the in vivo detection of fluorescent proteins. To demonstrate the benefits of FIM, we analyzed large groups of crawling Drosophila larvae. The number of analyzable locomotion tracks was increased by implementing a new software module capable of preserving larval identity during most collision events. This module is integrated in our new tracking program named FIMTrack which subsequently extracts a number of features required for the analysis of complex locomotion phenotypes. FIM enables high throughput screening for even subtle behavioral phenotypes. We tested this newly developed setup by analyzing locomotion deficits caused by the glial knockdown of several genes. Suppression of kinesin heavy chain (khc) or rab30 function led to contraction pattern or head sweeping defects, which escaped in previous analysis. Thus, FIM permits forward genetic screens aimed to unravel the neural basis of behavior.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drosophila / genetics
  • Drosophila / physiology*
  • Drosophila Proteins / genetics
  • Drosophila Proteins / physiology*
  • Female
  • Kinesin / genetics
  • Kinesin / physiology
  • Larva / genetics
  • Larva / physiology
  • Locomotion / genetics
  • Locomotion / physiology*
  • Male
  • Monomeric GTP-Binding Proteins / genetics
  • Monomeric GTP-Binding Proteins / physiology
  • Neuroglia / metabolism
  • RNA Interference
  • Reproducibility of Results
  • Video Recording / methods*

Substances

  • Drosophila Proteins
  • Kinesin
  • Monomeric GTP-Binding Proteins
  • Rab30 protein, Drosophila

Grant support

ST acknowledges a predoctoral fellowship of the Boehringer Ingelheim Fonds. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.