To understand brain functions, it is important to observe directly how multiple neural circuits are performing in living brains. However, due to tissue opaqueness, observable depth and spatiotemporal resolution are severely degraded . Here, we propose an optical brain clearing method for fluorescence microscopy, termed MAGICAL (magical additive glycerol improves clear alive luminance). MAGICAL enabled two-photon microscopy to capture vivid images with fast speed, at cortical layer V and hippocampal CA1 . Moreover, MAGICAL promoted conventional confocal microscopy to visualize finer neuronal structures including synaptic boutons and spines in unprecedented deep regions, without intensive illumination leading to phototoxic effects. Fluorescence emission spectrum transmissive analysis showed that MAGICAL improved transmittance of shorter wavelength light, which is vulnerable to optical scattering, thus unsuited for microscopy. These results suggest that MAGICAL would transparentize living brains via scattering reduction.
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| http://dx.doi.org/10.1016/j.isci.2020.101888 | DOI Listing |
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