We collected Salt Creek pupfish (Cyprinodon salinus salinus) from Salt Creek, Death Valley, California, in November 2009 and May 2010. The purpose of our study was to determine whether otoliths displayed interpretable marks that might be used for estimating age and growth. Otoliths exhibited alternating bands of opaque and translucent material. Kendall rank correlation between number of bands on otoliths and length of fish were high for two readers (τ = 0.65 and 0.79) and exact agreement between readers was 51%. Otoliths exhibited 0–5 bands, which provided evidence that longevity of Salt Creek pupfish likely is >1 year. Total length of fish collected in spring and autumn differed for fish with one and three bands on otoliths.