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Mica nanolayers have great potential for applications in the future flexible electronics due to their excellent flexibility and dielectric properties. Characterization of the adhesion behaviors of mica nanolayers is thus essential, as they would substantially affect the reliability of the electronic devices. In this work, a new delamination method is developed based on the optical microscopic nanomanipulation for measuring the adhesion energy of mica nanolayers. In particular, the tests are performed under different environmental conditions with relative humidity varying from 10% to 90% and temperature ranging from 25 to 300 °C. The average surface energy of mica nanolayers is 221.5 ± 25.4 mJ m−2 measured at the room temperature of 25 °C and decreases with the increase in relative humidity or temperature. The finding of the environmentally dependent adhesion is of importance for the design of flexible electronics products based on mica nanolayers. In this paper, a new method is introduced to measure adhesion of nanomaterials based on the bridge fabricated by nanomanipulation. The adhesion of mica nanolayer is investigated to demonstrate the advantage of this method, which could not be achieved by previous methods. Moreover, this method also fills the blank of studying the influence of environmental factors on the adhesion of nanomaterials. [ABSTRACT FROM AUTHOR]