A daylight fluorescence imager was deployed on an autonomous rover, Zoë, to detect life on the surface and shallow subsurface in regions of the Atacama Desert in Chile during field tests between 2003 and 2005. In situ fluorescent measurements were acquired from naturally fluorescing biomolecules such as chlorophyll and from specific fluorescent probes sprayed on the samples, targeting each of the four biological macromolecule classes: DNA, protein, lipid, and carbohydrate. RGB context images were also acquired. Preparatory reagents were applied to enhance the dye probe penetration and fluorescence intensity of chlorophyll. Fluorescence imager data sets from 257 samples were returned to the Life in the Atacama science team. A variety of visible life forms, such as lichens, were detected, and several of the dye probes produced signals from nonphotosynthetic microorganisms.