Light attenuation in marine ecosystems can limit primary production and determine the species composition and abundance of primary producers. In Florida Bay, the importance of understanding the present light environment has heightened as major upstream water management restoration projects have been proposed and some are already being implemented. We analyzed a 2-yr (2001–2003) data set of the light attenuation coefficient (Kt) and its principal components (water, chromophoric dissolved organic matter [CDOM], tripton, phytoplankton) obtained at 40 stations within Florida Bay, calibrated synoptic underway data to produce high spatial resolution maps, examined the potential for light limitation, and quantified the individual effect of each component upon light attenuation. Tripton was the dominant component controlling light attenuation throughout Florida Bay, whereas the contribution of chlorophylla and CDOM to Kt was much smaller in all regions of Florida Bay. It was possible to accurately estimate the light attenuation coefficient from component concentrations, using either a mechanistic or a statistical model with root mean square errors of 0.252 or 0.193 m−1, respectively. Compared to other estuaries, Florida Bay had the lowest overall Kt and the greatest relative contribution from tripton. Comparing the recent data to a study of Florida Bay’s light environment conducted in 1993–1994, we found that overall water clarity in the Bay increased significantly, indicated by a nearly 3-fold decrease in Kv as a result of lower tripton concentrations, although the percent contribution of each of the components to Kt is unchanged. Only the northwest corner of Florida Bay, an area comprised of approximately 8% of the Bay’s total area, was found on average to have sufficient light attenuation to limit the growth of seagrasses. This is much less extensive than in 1993–1994, when seagrass growth was potentially limited by light at over 50% of the stations sampled.