Optical Detection and Imaging of Nonfluorescent Matter at the Single-Molecule/Particle Level

Since the first optical detection of single molecules in 1989, single-molecule spectroscopy has developed rapidly and been widely applied in many areas. However, the vast majority of matter is extremely inefficient at emitting photons in our physical world, which seriously limits the applications of optical methods based on photoluminescence. In addition to indirect detection by fluorescence labeling, many efforts have been made to directly image nonfluorescent matter at the single-particle or single-molecule level in different ways based on the absorption or scattering interaction between light and matter. Herein, we review five popular methods for imaging nonfluorescent particles/molecules, including dark-field microscopy (DFM), surface plasmon resonance microscopy (SPRM), surface enhanced Raman microscopy (SERM), interferometric scattering microscopy (iSCAT), and photothermal microscopy (PTM). After summarizing the principles and applications of these methods, we compare the advantages and disadvantages of each method and describe further potential development and applications.