Fluorescence correlation spectroscopy (FCS) is a powerful technique for quantification of molecular dynamics, and it has been widely applied in diverse fields, e.g., biomedicine, biophysics, and chemistry. By time-correlation of the fluorescence fluctuations induced by molecules diffusing through a focused light, FCS can quantitatively evaluate the concentration, diffusion coefficient, and interaction of the molecules in vitro or in vivo. In this review, the basic principle and implementation of FCS are introduced. Then, the advances of FCS variants are reviewed, covering dual-color FCCS, multi-focus FCS, pair correlation function (pCF), scanning FCS, focus-reduced FCS, SPIM-FCS, and inverse-FCS. Besides, the applications of FCS are demonstrated with the measurement of local concentration, hydrodynamic radius, diffusion coefficient, and the interaction of different molecules. Lastly, a discussion is given by summarizing the pros and cons of different FCS techniques, as well as the outlooks and perspectives of FCS.

Panoramic and long-term observation of nanosized organelle dynamics and interactions with high spatiotemporal resolution still hold great challenge for current imaging platforms. In this study, we propose a live-organelle imaging platform, where a flat-fielding quantitative phase contrast microscope (FF-QPCM) visualizes all the membrane-bound subcellular organelles, and an intermittent fluorescence channel assists in specific organelle identification. FF-QPCM features a high spatiotemporal resolution of 245 nm and 250 Hz and strong immunity against external disturbance. Thus, we could investigate several important dynamic processes of intracellular organelles from direct perspectives, including chromosome duplication in mitosis, mitochondrial fusion and fission, filaments, and vesicles’ morphologies in apoptosis. Of note, we have captured, for the first time, a new type of mitochondrial fission (entitled mitochondrial disintegration), the generation and fusion process of vesicle-like organelles, as well as the mitochondrial vacuolization during necrosis. All these results bring us new insights into spatiotemporal dynamics and interactions among organelles, and hence aid us in understanding the real behaviors and functional implications of the organelles in cellular activities.

Super-resolution optical microscopy, which gives access to finer details of objects, is highly desired for fields of nanomaterial, nanobiology, nanophotonics, etc. Many efforts, including tip optimization and illumination optimization etc., have been made in both near-field and far-field super-resolution microscopy to achieve a spatial resolution beyond the diffraction limit. The development of vector light fields opens up a new avenue for super-resolution optical microscopy via special illumination modes. Cylindrical vector beam (CVB) has been verified to enable resolution improvement in tip-scanning imaging, nonlinear imaging, stimulated emission depletion (STED) microscopy, subtraction imaging, superoscillation imaging, etc. This paper reviews recent advances in CVB-based super-resolution imaging. We start with an introduction of the fundamentals and properties of CVB. Next, strategies for CVB based super-resolution imaging are discussed, which are mainly implemented by tight focusing, depletion effect, plasmonic nanofocusing, and polarization matching. Then, the roadmap of super-resolution imaging with CVB illumination in the past two decades is summarized. The typical CVB-based imaging techniques in fields of both near-field and far-field microscopy are introduced, including tip-scanning imaging, nonlinear imaging, STED, subtraction imaging, and superoscillation imaging. Finally, challenges and future directions of CVB-illuminated super-resolution imaging techniques are discussed.

Function expansion of fiber sensor is highly desired for ultrasensitive optical detection and analysis. Here, we present an approach of multifunctional fiber sensor based on Fano resonances and localized surface plasmon resonance (LSPR) excited via cylindrical vector beam with ability of refractive index (RI) sensing, nano-distance detection, and surface enhanced Raman spectroscopy (SERS). Silver (Ag)-nanocube modified microfiber is theoretically proved to enable to detect RI of the nearby solids and gases based on Fano resonances with a sensitivity of 128.63 nm/refractive index unit (RIU) and 148.21 nm/RIU for solids and gases, respectively. The scattering spectrum of the Ag nanocube has the red-shift response to the varies of the nano-distance between the nanocube and the nearby solid, providing a detection sensitivity up to 1.48 nm (wavelength)/nm (distance). Moreover, this configuration is theoretically verified to have ability to significantly enhance electric field intensity. Radially polarized beam is proved to enhance the electric field intensity as large as 5 times in the side-face configuration compared with linear polarization beam. This fiber-based sensing method is helpful in fields of remote detection, multiple species detection, and cylindrical vector beam-based detection.