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FIM Imaging and FIMtrack: Two New Tools Allowing High-throughput and Cost Effective Locomotion Analysis

The analysis of neuronal network function requires a reliable measurement of behavioral traits. Since the behavior of freely moving animals is variable to ...

Westfälische Wilhelms-Universität Münster

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.

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

Quantitative analysis of behavioral traits requires precise image acquisition and sophisticated image analysis to detect subtle locomotion phenotypes. In the past, we have established Frustrated Total Internal Reflection (FTIR) to improve the measurability of small animals like insects. This FTIR-based Imaging Method (FIM) results in an excellent foreground/background contrast and even internal organs and other structures are visible without any complicated imaging or labeling techniques. For example, the trachea and muscle organizations are detectable in FIM images. Here these morphological details are incorporated into phenotyping by performing cluster analysis using histogram-based statistics for the first time. We demonstrate that FIM enables the precise quantification of locomotion features namely rolling behavior or muscle contractions. Both were impossible to quantify automatically before. This approach extends the range of FIM applications by enabling advanced automatic phenotyping for particular locomotion patterns.

Nils Otto

Institute of Neuro and Behavioral Biology

Westfälische Wilhelms-Universität Münster

FIM Imaging and FIMtrack: Two New Tools Allowing High-throughput and Cost Effective Locomotion Analysis

Nils Otto

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Institute of Neuro and Behavioral Biology

Westfälische Wilhelms-Universität Münster

FIM Imaging and FIMtrack: Two New Tools Allowing High-throughput and Cost Effective Locomotion Analysis

Institute of Neuro and Behavioral Biology