This protocol outlines the Clear+ Passive method to achieve optical transparency in rat brain hemispheres, indicating its potential for imaging and immunolabeling.
This protocol describes the implementation of the Clear+ Passive brain-clearing method for the George Lab, with additional notes on reagent sources and minor workflow adaptations. The protocol steps are directly transcribed from the LifeCanvas Technologies documentation (LifeCanvas Wiki). Original authorship and credit belong entirely to LifeCanvas Technologies. This protocol outlines the Clear+ Passive procedure for clearing whole rat brain hemispheres to achieve optical transparency compatible with light-sheet imaging and immunolabeling. Following SHIELD stabilization, delipidation is performed, followed by immunolabeling and final refractive index matching in EasyIndex (RI ≈ 1.52). The Clear+ Passive method provides a gentle, detergent-based approach designed to preserve protein integrity and tissue morphology in large samples. This implementation remains a work in progress and has not yet been imaged to verify clearing and labeling outcomes in rat brain hemispheres.
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Mosquera et al. (2025) studied this question.
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