Journal
FRONTIERS IN NEUROANATOMY
Volume 12, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fnana.2018.00067
Keywords
tissue optical clearing; whole-brain imaging; green fluorescent protein; fluorescence preservation; uDISCO; a-uDISCO
Categories
Funding
- National Key Research and Development Program of China [2017YFA0700501]
- National Natural Science Foundation of China [31571002, 81701354]
- Science Fund for Creative Research Group of China [61721092]
- China Postdoctoral Science Foundation [2017M612463, 2018T110772]
- Fundamental Research Funds for the Central Universities, HUST [2018KFYXKJC026]
- WNLO
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Tissue optical clearing techniques provide alternative approaches for imaging large-volume specimens. uDISCO, an organic-solvent-based method, stands out from the enormous array of available optical clearing methods by achieving whole-brain imaging with high transparency, size reduction and fluorescence preservation. In this study, we aimed to modify the uDISCO protocol to achieve better fluorescence preservation and to thereby further improve its optical imaging quality. First, we determined the optimal pH value for optimized uDISCO, termed a-uDISCO (alkaline pH-based uDISCO). Then, we compared fluorescence preservation between a-uDISCO and uDISCO. In addition, we validated the clearing performance of the optimized method according to several parameters, including tissue transparency, size changes, and the maintenance of cell morphology. Finally, we demonstrated that a-uDISCO enabled the high-quality brain-wide visualization of neuronal structures. This method potentially provides a better alternative for high-throughput imaging of samples with low-level fluorescence protein expression or for archiving and repetitive revisiting of rare samples.
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