Cycling has a range of benefits as is recognised by national and international policies aiming to increase cycling rates. Darkness acts as a barrier to people cycling, with fewer people cycling after-dark when seasonal and time-of-day factors are accounted for. This paper explores whether road lighting can reduce the negative impact of darkness on cycling rates. Changes in cycling rates between daylight and after-dark were quantified for 48 locations in Birmingham, United Kingdom, by calculating an odds ratio. These odds ratios were compared against two measures of road lighting at each location: 1) Density of road lighting lanterns; 2) Relative brightness as estimated from night-time aerial images. Locations with no road lighting showed a significantly greater reduction in cycling after-dark compared with locations that had some lighting. A nonlinear relationship was found between relative brightness at a location at night and the reduction in cyclists after-dark. Small initial increases in brightness resulted in large reductions in the difference between cyclist numbers in daylight and after-dark, but this effect reached a plateau as brightness increased. These results suggest only a minimal amount of lighting can promote cycling after-dark, making it an attractive mode of transport year-round.