Abstract
Single-molecule detection has become a unique and indispensable tool for the study of molecular motions and interactions at the single-molecule level. Unlike ensemble measurement where the information is averaged, single-molecule analysis yields invaluable information on both the individual molecular properties and their microenvironment. Among the various technologies for the detection of single molecules, the detection with optical methods has many advantages in terms of its high sensitivity, electrical passiveness, and robustness. The recent advances in the engineering of either the excitation light or the solution of the molecules have paved the way for enhanced single-molecule detection. We present recent developments and future perspectives for single-molecule detection in the following three regimes: on a dry surface, in solutions at ultralow concentrations, and in solutions at native physiological concentrations.
Highlights
The goal of sensing in biology is to have the capability to study a wide variety of single-molecule behaviors
Optical single-molecule detection is one such technology that enables the study of molecular motion and interaction at the single-molecule level, and it has become an indispensable tool in many applications, such as diagnostics,[1] DNA sequencing,[2,3] and molecular biology.[4]
This review mainly focuses on the single-molecule detection in the following three regimes: (1) on a dry surface, (2) in solutions at ultralow concentrations, and (3) in solutions at native physiological concentrations
Summary
The goal of sensing in biology is to have the capability to study a wide variety of single-molecule behaviors. This review mainly focuses on the single-molecule detection in the following three regimes: (1) on a dry surface, (2) in solutions at ultralow concentrations, and (3) in solutions at native physiological concentrations. The detection in solutions at ultralow concentrations requires the development of ultrasensitive technology to distinguish the few molecules in the solution. The remainder of this review article is outlined as follows: Sec. 2 will describe the techniques for the advanced detection of single molecules on a surface and at low concentrations; Sec. 3 will introduce methods that have been applied to overcome the diffusion limit or the detection-time issue; Sec. 4 will review the recent developments for the optical detection of single molecules at native physiological concentrations; and Sec. 5 will conclude with a discussion of the perspectives for future developments in this promising field
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