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. 2016 May 26;20(Suppl 1):S15–22. doi: 10.5213/inj.1632630.315

Fig. 2.

Fig. 2.

Comparison of polymer-based methods. (A) CLARITY tissue-polymer system and processing time. Hydrogel monomer mixture contains 4% PFA, 4% acrylamide, and 0.05% bis-acrylamide. After polymerization, cross-linking between tissue and hydrogel monomers occurs. The entire processing time to clarify the whole brain of adult mouse is over 2 weeks. (B) PACT tissue-polymer system and processing time. This method uses a thick-sliced, fixed sample and 4% acrylamide solution (hydrogel monomer solution), and the lipid is passively diffused out from the tissue. The entire processing time to clarify adult mouse brain slices (2-to 3-mm thickness) is over 1 week. (C) ACT tissue-polymer system and processing time. Fixed whole brain is incubated in 4% acrylamide solution. After polymerization, the tissue has a less complex tissue-hybrid network formation than CLARITY, and lipid is removed by electrophoretic tissue clearing within 6 hours. CLARITY, clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization-compatible tissue-hydrogel; PFA paraformaldehyde; PACT, passive clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization-compatible tissue-hydrogel; ACT, active clarity technique; SDS, sodium dodecyl sulfate .