Massively parallel C. elegans tracking provides multi-dimensional fingerprints for phenotypic discovery

Abstract

The WF-NTP extends significantly the power of existing automated platforms by combining enhanced optical imaging techniques with an advanced software platform. We anticipate that this approach will further extend the scope and utility of C. elegans as a model organism.

Automated Behavioral Analysis of Large <em>C. elegans</em> Populations Using a Wide Field-of-view Tracking Platform

“…The ease of operating, multiparametric analysis, and high throughput of the illustrated WF-NTP protocol (Figures 1 and 2) makes it possible to determine very small changes in worm behavior in a very accurate manner. The imaging platform is based on custom-made opto-mechanics, and it can be assembled using a 6 MP monochrome camera combined with 16 mm focal length high-resolution lens for 1'' sensor, illuminated with a 8'' by 8'' white backlight (see Table of Materials and also reference 36 for additional details). The associated WF-NTP software is written in Python and was developed to run on Windows platforms.…”

“…The results that have been achieved so far in AD 15,34,35 and PD 18 drug discovery demonstrate the importance of wide field-of-view data acquisition to greatly increase the numbers of animals that can be monitored in a single experiment, significantly decreasing the experimental errors and greatly improving the statistical validity of the studies. Based on these results, we anticipate that the WF-NTP protocol, which we made readily available for the community 36 , will significantly extend the use of C. elegans. Specific videos and selected frames can be selected in the GUI for analysis, and a pixel conversion factor can be inserted, after which the analysis is carried out with one of the two given tracking algorithms.…”

“…In order to expand further the process of automation to the large cohorts of animals needed for statistically significant measurements, we have recently developed a wide field-of-view nematode tracking platform (WF-NTP) 15,34,35,36 , which enables the simultaneous investigation of multiple phenotypic readouts on very large worm populations, a key factor in statistically relevant phenotypical detection The associated open-source software is written in Python, which is also required to operate it and is completely customizable. A graphical user interface (GUI) is also provided to enable users to adopt this technology.…”

“…The key steps during the behavioral screening are the timings of the recording and any worm handling steps (e.g., the transfer of worms from the NGM plates to the tracking platform). The protocol described here is an example designed to screen the worms for up to 9 days during the adult lifespan; however, this protocol can be easily adapted to screen as many time points as the user desires, e.g., 18 consecutive days 36 . Part 4 then illustrates the application of the protocol to deliver protein molecules (e.g., antibodies and molecular chaperones) into C. elegans, and shows how the protocol illustrated in Parts 1-3 can be easily customized, depending on the desired application.…”

An open source platform for analyzing and sharing worm behavior data

“…Several worm trackers have been previously described [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] . Most of the trackers either extract detailed postural information of one worm at a time or track multiple worms but extract only coarser-grained features such as centroid speed or orientation.…”