Using Tampa Bay, FL as an example, we explored the potential for using MODIS medium-resolution bands (250- and 500-m data at 469-, 555-, and 645-nm) for estuarine monitoring. Field surveys during 21–22 October 2003 showed that Tampa Bay has Case-II waters, in that for the salinity range of 24–32 psu, (a) chlorophyll concentration (11 to 23 mg m−3), (b) colored dissolved organic matter (CDOM) absorption coefficient at 400 nm (0.9 to 2.5 m−1), and (c) total suspended sediment concentration (TSS: 2 to 11 mg L−1) often do not co-vary. CDOM is the only constituent that showed a linear, inverse relationship with surface salinity, although the slope of the relationship changed with location within the bay. The MODIS medium-resolution bands, although designed for land use, are 4–5 times more sensitive than Landsat-7/ETM+ data and are comparable to or higher than those of CZCS. Several approaches were used to derive synoptic maps of water constituents from concurrent MODIS medium-resolution data. We found that application of various atmospheric-correction algorithms yielded no significant differences, due primarily to uncertainties in the sensor radiometric calibration and other sensor artifacts. However, where each scene could be groundtruthed, simple regressions between in situ observations of constituents and at-sensor radiances provided reasonable synoptic maps. We address the need for improvements of sensor calibration/characterization, atmospheric correction, and bio-optical algorithms to make operational and quantitative use of these medium-resolution bands.