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Since their invention in 1986 by Arthur Ashkin and colleagues, opticaltweezers have become an essential tool in several fields of physics,spectroscopy, biology, nanotechnology, and thermodynamics. In this Tutorial, weprovide a primer on how to calibrate optical tweezers and how to use them foradvanced applications. After a brief general introduction on optical tweezers,we focus on describing and comparing the various available calibrationtechniques. Then, we discuss some cutting-edge applications of optical tweezersin a liquid medium, namely to study single-molecule and single-cell mechanics,microrheology, colloidal interactions, statistical physics, and transportphenomena. Finally, we consider optical tweezers in vacuum, where the absenceof a viscous medium offers vastly different dynamics and presents newchallenges. We conclude with some perspectives for the field and the futureapplication of optical tweezers. This Tutorial provides both a step-by-stepguide ideal for non-specialists entering the field and a comprehensive manualof advanced techniques useful for expert practitioners. All the examples arecomplemented by the sample data and software necessary to reproduce them.

Optical Tweezers: A Comprehensive Tutorial from Calibration to Applications