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Tissue clearing methods are growing in popularity due to their improved ability to image deeper while maintaining fluorescence of fluorophores. Although there are many tissue clearing methods present for a long time, new tissue clearing methods were developed to improve the depth of imaging while reducing fluorescence quenching. In broad, the tissue clearing methods are classified into water-based and organic solvent-based methods. Water-based methods (CLARITY, PACT-PARS, CUBIC, Scale) take a long time for clearing while good at preserving fluorescent proteins. On the other hand, organic solvent-based methods (BABB, 3DISCO, FluoClear BABB, Ethanol-ECi, and PEGASOS) achieve tissue clearing in a short amount of time but do not preserve well protein-based fluorescence. Among all these methods, 3DISCO, PACT and CUBIC were reported to be superior for deep tissue imaging. Although all three methods achieve high transparency, 3DISCO is much faster (hours to days instead of weeks). Water-based clearing methods (TDE, Clear See and methods based on Scale) were optimised for plant tissues. However, 3DISCO (or its variants) are yet to be adapted for plants. 3DISCO, being an organic solvent-based method, allows deeper imaging and faster clearing times. In this study, we are trying to adapt 3DISCO to various plant species.
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