Developing the aggregation indcued emission (AIE) nanodots as alternates to the conventional inorganic quantum dots is a contemporary research interest towards assorted applications. Herein, we have designed and synthesized a tripodal structured pyrene Schiff base (PS) and it is characterized by 1H NMR, 13C NMR and ESI‐MS techniques. The developed PS exhibit an excellent AIE property in THF:water (10:90) ratio. The formed PS aggregates were meticulously characterized by HR‐SEM, HR‐TEM and AFM measurements. The results reveal that the PS aggregates are spherical in shape with an average diameter of ∼ 1 nm. Hence, the PS aggregates are expressed as AIE nanodots. Intriguingly, the AIE nanodots has high fluorescence lifetime (4 ns) and quantum yield of 54 %, which are adequate to compete with the commercial fluorophores. In addition, molecular structure dependent nano shapes have also been investigated. The PS based AIE nanodots were successfully utilized as a fluorescent probe for several applications such as in‐vitro bioimaging, detection of picric acid and photoinduced electron transfer studies. The confocal fluorescence images of nanodots in HeLa and HEK live cells exhibit higher fluorescence signal which clearly reveal that AIE nanodots can be used as a promising probe for in‐vitro bio‐imaging application. Moreover, the AIE nanodots are employed to detect the picric acid through fluorescence quenching method. Towards the commercial application, the low‐cost fluorescent test strips have also been developed for the detection of picric acid. We have also investigated the electron transfer nature of AIE nanodots with benzyl viologen towards the energy related applications.