How to Identify the "LIVE/DEAD" States of Microbes Related to Biosensing

In this work, we fabricated a microbial biosensor with long-term stability, which relied on microbial activity. Activity of the microbe was commonly estimated by LIVE/DEAD assay and the propidium iodide (PI)-stained one was judged as dead. Herein, we proposed the utilization of a physiological state of microbes, which was neither live nor dead but between them. In this state, microbes represented a high PI-stained ratio but still had catalytic ability. This microbial state was obtained by forming the biofilm under the conditions of poor nutrition and low temperature. Thus, the dividing and proliferating ability of the microbes in the biofilm was weak, which was beneficial for long-term stability. This mechanism was further confirmed by the biosensors made from multifarious substrate materials, including graphene-based gel, biomass-based gel, graphite felt, and poly(vinyl chloride). This biosensor was applied to water pollution monitoring in the laboratory for 2 years and then was integrated into a multiparameter water quality monitoring station on a local lake for 2.5 years.