The stressful conditions of today-life make it urgent the timely prevention and treatment of many physiological and psychological disorders related to stress. According to the significant progress made in the near future, rapid, accurate, and on-spot measurement of cortisol hormone as a dominant stress biomarker using miniaturized digital devices is not far from expected. With a special potency in the fields of diagnosis and healthcare monitoring, aptamer-mediated biosensors (aptasensors) are promising for the quantitative monitoring of cortisol levels in the different matrices (sweat, saliva, urine, cerebrospinal fluid, blood serum, etc.). Accordingly, this in-depth study reviews the superior achievements in the aptasensing strategies to detect cortisol hormone with the synergism of diverse two/three dimensional nanostructured materials, enzymatic amplification components, and antibody motifs. The represented discussions offer a universal perspective to achieve lab-on-chip aptasensing arrays as future user-friendly skin-patchable electronic gadgets for on-site and real-time quantification of cortisol levels.