Cyanobacterial mat communities are the main drivers of primary productivity in the McMurdo Dry Valleys, Antarctica. These microbial communities form laminar mats on desert pavement surfaces adjacent to glacial meltwater streams, ponds, and lakes. The low-density nature of these communities and their patchy distribution make assessments of distribution, biomass, and productivity challenging. We used satellite imagery coupled with in situ surveying, imaging, and sampling to systematically estimate microbial mat biomass in selected wetland regions in Taylor Valley, Antarctica. On January 19th, 2018, the WorldView-2 multispectral satellite acquired an image of our study areas, where we surveyed and sampled seven 100 m² plots of microbial mats for percent ground cover, ash-free dry mass (AFDM), and pigment content (chlorophyll-a, carotenoids, and scytonemin). Multispectral analyses revealed spectral signatures consistent with photosynthetic activity (relatively strong reflection at near-infrared wavelengths and relatively strong absorption at visible wavelengths), with average normalized difference vegetation index (NDVI) values of 0.09 to 0.28. Strong correlations of microbial mat ground cover (R² = 0.84), biomass (R² = 0.74), chlorophyll-a content (R² = 0.65), and scytonemin content (R² = 0.98) with logit transformed NDVI values demonstrate that satellite imagery can detect both the presence of microbial mats and their key biological properties. Using the NDVI—biomass correlation we developed, we estimate carbon (C) stocks of 21,715 kg (14.7 g C m⁻²) in the Canada Glacier Antarctic Specially Protected Area, with an upper and lower limit of 74,871 and 6312 kg of C, respectively.