In order to recognize single worms and have the ability to track them, the organism silhouettes need to be recognized by SMART software. When worms are too close or stacked, the algorithm will not be able to separate them. This particular issue is important when dealing with dead worms. For example, if you decide to treat worms in liquid and later measure them on a Petri dish, you should pay attention to how you pipette them or how you disperse them.
The easy way to do it is to pass worms into separate drops (6 microliters each, for example), so the drop volume will be enough to disperse the worms by surface tension when it falls onto the plate. This trick is effective most of the time. However, you need to visualize it under the dissecting scope to confirm.
An alternative methodology to separate dead worms for imaging and counting is to employ a small brush. By softly passing the brush above the surface, it will disperse the worms.
