We can't find the internet
Attempting to reconnect
Something went wrong!
Hang in there while we get back on track
Fluorescence radial fluctuation enables two-photon super-resolution microscopy
Summary
Researchers applied super-resolution radial fluctuation analysis to two-photon microscopy to achieve high-resolution imaging deep within brain tissue. The technique achieved spatial resolution comparable to structured illumination microscopy at depths of several hundred micrometers and enabled the first in vivo super-resolution imaging of the fifth layer of the cerebral cortex, offering an accessible upgrade for existing two-photon microscope systems.
Despite recent improvements in microscopy, it is still difficult to apply super-resolution microscopy for deep imaging due to the deterioration of light convergence properties in thick specimens. As a strategy to avoid such optical limitations for deep super-resolution imaging, we focused on super-resolution radial fluctuation (SRRF), a super-resolution technique based on image analysis. In this study, we applied SRRF to two-photon microscopy (2P-SRRF) and characterized its spatial resolution, suitability for deep observation, and morphological reproducibility in real brain tissue. By the comparison with structured illumination microscopy (SIM), it was confirmed that 2P-SRRF exhibited two-point resolution and morphological reproducibility comparable to that of SIM. The improvement in spatial resolution was also demonstrated at depths of more than several hundred micrometers in a brain-mimetic environment. After optimizing SRRF processing parameters, we successfully demonstrated in vivo high-resolution imaging of the fifth layer of the cerebral cortex using 2P-SRRF. This is the first report on the application of SRRF to in vivo two-photon imaging. This method can be easily applied to existing two-photon microscopes and can expand the visualization range of super-resolution imaging studies.
Sign in to start a discussion.
More Papers Like This
In vivo super-resolution of the brain – How to visualize the hidden nanoplasticity?
Researchers reviewed how super-resolution fluorescence microscopy techniques — which allow scientists to image structures smaller than what conventional light microscopes can resolve — are being used to study the nanoscale structure and plasticity of brain synapses in living mice. These imaging advances help reveal how tiny changes in brain connections relate to learning and memory, using "nanoplasticity" in its neurological sense rather than as a reference to plastic pollution.
Imaging dendritic spines in the hippocampus of a living mouse by 3D-STED microscopy
Researchers extended 3D STED super-resolution microscopy to image dendritic spines in the hippocampus of living mice, achieving nanoscale resolution in three dimensions within deep brain tissue and opening new possibilities for studying synaptic structures in vivo.
Cortex-Wide, Cellular-Resolution Volumetric Imaging with a Modular Two-Photon Imaging Platform
Researchers developed Meso2P, a modular two-photon imaging platform that achieves cortex-wide, cellular-resolution volumetric imaging by decoupling excitation and detection using a lateral paraboloid fluorescence collector. The system sustains an effective numerical aperture of 0.87 over a contiguous 6x6 mm2 field-of-view at 7.67 Hz, enabling mapping of neuronal activity across the entire cortex at single-cell resolution.
Cortex-Wide, Cellular-Resolution Volumetric Imaging with a Modular Two-Photon Imaging Platform
This paper presents Meso2P, a new two-photon microscope capable of imaging the entire mouse cortex at single-cell resolution — a significant advance in neuroscience instrumentation. Among its demonstrated applications is the ability to track the distribution of micro- and nanoplastic particles in living brain tissue in real time. While primarily a neuroscience tool, its capacity to visualize nanoplastics in the brain non-invasively could become valuable for directly studying how plastic particles move through and accumulate in neural tissue.
Editorial: 15 years of Frontiers in Cellular Neuroscience: super-resolution microscopy in the healthy and the injured brain
This editorial introduces a research collection on super-resolution microscopy techniques applied to the healthy and injured brain, highlighting how methods that surpass the diffraction limit of classical fluorescence microscopy are revealing new insights into synaptic organization and cellular pathology. The collection covers advances in both imaging hardware and computational image analysis relevant to neuroscience research.