Intrinsic Burst-Blinking Nanographenes for Super-Resolution Bioimaging
Nanographenes, synthesized with biocompatibility and functionalization groups, hold great promise as fluorophores for optical super-resolution microscopy. Such molecules exhibit excellent photo-blinking properties across diverse environments, including air, phosphate-buffered saline, and acidic or basic conditions. In our study, we demonstrated their applicability in materials imaging, live-cell imaging, and functional analysis of neurons at the nanometer scale.
Single-molecule localization microscopy (SMLM) is a groundbreaking imaging technique that enables biologists to visualize subcellular structures with an unprecedented spatial resolution of approximately 20 nanometers—a feat unattainable using conventional fluorescence microscopy. However, the current fluorophores employed in SMLM are highly sensitive to environmental conditions, which limits their applicability.
In our study, we present a significant advancement: the synthesis and functionalization of intrinsic blinking nanographenes. These nanographenes exhibit remarkable photo-blinking properties across various imaging conditions. Using functionalized nanographenes, we successfully demonstrated their super-resolution imaging applications from material science to cell biology. For example, we successfully imaged amyloid fibrils both in air and in varying pH solutions. In live cells under physiological conditions, we captured lysosome dynamics without any additional additives. Furthermore, we demonstrated the application of nanographenes in SMLM for neuroscience
research, opening up new avenues for molecular neuroscience studies. These findings underscore the immense potential of functionalized nanographenes as intrinsic burst-blinking fluorophores, paving the way for expanded super-resolution microscopy applications.