C. elegans Movement Tracking: A Method to Assess Locomotion in Worms

Abstract

Source: Davies, A. G. et al. An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans. J. Vis. Exp. (2015).

This video describes the use of video and tracking software to record worm movement in an arena.  The example protocol measures the effect of ethanol exposure on worm locomotion.

Protocol

The following protocol is an excerpt from Davies et al, An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans, J. Vis. Exp. (2015). Steps to Perform on the Day before the Assay Pick L4 stage worms to fresh nematode growth medium (NGM) plates seeded with…

Materials

C. elegans strains	Caenorhabditis Genetics Center
60 x 15 mm Petri plates, triple vented	Greiner Bio-One	628161	Other plate brands will suffice.
NGM agar	Various		NaCl (3 g/L), agar (17 g/L), peptone (2.5 g/L), 1 ml cholesterol (5 mg/ml in ethanol), 1 ml (1 M) MgSO4, 1 ml (1 M) CaCl2, 25 ml (1 M) KPO4, pH=6, 975 ml H2O
Forceps	Various		e.g., Fisher Scientific #10300
37 °C Incubator	Various		For drying agar
Copper rings	Plumbmaster	STK#35583 (48 cap thread gasket)	1.6 cm inner diameter, 1.8 cm outer diameter copper rings
100% ethanol	Various
Parafilm M	Bemis	PM996
CCD camera	QImaging	RET-4000R-F-M-12	This camera has a large field of view.
Stereomicroscope with C-mount and 0.5X objective	Leica	MZ6	Discontinued model, M60 is current equivalent.
Light source	Schott	A08923	3”x3”  backlight for even illumination across the field of view
Imaging and tracking software	Media Cybernetics	ImagePro-Plus v6.0-6.3	Newer versions of the software have tracking functions.