Journal
ISCIENCE
Volume 24, Issue 1, Pages -Publisher
CELL PRESS
DOI: 10.1016/j.isci.2020.101888
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Funding
- Brain/MIND'' from the Japan Agency for Medical Research and Development (AMED) [20dm0207078]
- MEXT/JSPS KAKENHI from the Japan Society for the Promotion of Sciences (JSPS) in the Ministry of Education, Culture, Sports, Science and Technology (MEXT) [JP15H05953, JP20H00523, JP20H05669]
- Network Joint Research Center for Materials and Devices'' in the MEXT
- Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials'' in the MEXT
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MAGICAL is an optical brain clearing method that enhances the performance of in vivo fluorescence microscopy, allowing for vivid imaging of deeper regions in living brains. It improves transmittance of shorter wavelength light, reducing optical scattering and making living brains more transparent.
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 in vivo. Here, we propose an optical brain clearing method for in vivo 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 in vivo. 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 in vivo transmittance of shorter wavelength light, which is vulnerable to optical scattering, thus unsuited for in vivo microscopy. These results suggest that MAGICAL would transparentize living brains via scattering reduction.
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