There is growing interest in monitoring soil biological health to complement the traditional evaluation of soil physical and chemical characteristics in agricultural fields. Activity of soil microorganisms mediates many essential soil processes that affect fertility, and, therefore, essential to the successful adoption of precision agriculture. However, there are technical limitations to cost-effective monitoring of spatial and temporal dynamics of soil biological activity across agricultural landscapes. This paper summarizes three consecutive studies on in situ measurement of soil biological activity. The first study reveals spatial heterogeneity of microbial population growth in three agricultural fields using bio-films. In the second study, microbiological activity was analyzed using a substrate-induced respiration technique. This technique was evaluated through a series of soil toxicity experiments that involved a comparison of fresh and autoclaved soil samples. Finally, the aim of the third study was to develop a portable instrumented system to evaluate carbon dioxide concentrations in soil by extracting air stored within the soil pores. This instrument was tested under various conditions to quantify the effects of soil moisture, compaction and presence of glucose (artificially increased microbial respiration). Optimization of the discussed techniques will allow for detailed mapping of these indices of soil biological health and their interactions with the physical and chemical environment at any specific point in time.