Unlike most darters, the Slough Darter inhabits quiet lowland pools having muddy bottoms. It prefers small, shaded streams and overflow ponds where there is leaf litter or aquatic vegetation. They tolerate occasional muddy water but always avoid strong currents.
This darter spawns in April and May in Kansas. The female attaches eggs to a thin plant stem or twig in a single row. The male follows to fertilize each egg as it is laid. They feed on larval midges, mayflies, and microcrustaceans.
Slough Darters are known in Kansas from only three streams: Cow Creek, a tributary to the Spring River; Fly Creek, a tributary to the Neosho River; and Elk River.