Year: 2026
Type: preprint
Abstract: Two-photon excitation (2PE) microscopy reaches deep into tissue but remains diffraction-limited, and existing 2PE-STED and 2PE-RESOLFT schemes still rely on single-photon depletion or photoswitching. Here we present 2P-RESOLFT microscopy, in which excitation and reversible photoswitching are both two-photon-driven. Using the positive photoswitchable protein Padron2, we achieved 205-nm lateral resolution and resolved previously unreported membrane protrusions of β cells inside pancreatic pseudoislets.
Institutions: The University of Osaka, National Institutes of Natural Sciences, The Graduate University for Advanced Studies, SOKENDAI, National Institute for Physiological Sciences, The Exploratory Research Center on Life and Living Systems, Institute for Open and Transdisciplinary Research Initiatives, Hamamatsu University, Hamamatsu University School of Medicine, Tohoku University, Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Ritsumeikan University
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
Funders: Japan Agency for Medical Research and Development, Japan Society for the Promotion of Science, Japan Science and Technology Agency, Core Research for Evolutional Science and Technology, National Institute for Physiological Sciences, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences
Awards: G1021988992, Development of highly-biocompatible super-resolution fluorescence polarization imaging, G1354701201, In vivo trans-scale visualization and analysis of mouse brain and neural functions through nonlinear optical processes, 多細胞組織観察に資する非線形超解像蛍光顕微鏡の開発, G2346724487, G2612887765, G2931199209, G2943184455, 生体深部の超解像イメージング法「2光子RESOLFT」の開発と応用, 長期記憶を司る局所翻訳システムの三次元超解像2光子イメージング解析, G3903484084, Development of a method for super-resolved chromophore-assisted laser inactivation, 超解像「生理機能」イメージング法の開発と細胞状態解析への応用, Advanced Bioimaging Support, Comprehensive Visualization of Intracellular Structure Through Integration of Super-resolution Imaging and Quantitative Phase Imaging, G6513824541, G6687149700, Development and Applications of Cryogenic Molecular Functional Imaging, G7043198086, G728880209
OpenAlex ID: W7213638603