Clear^T2 clears tissue with fluorescent proteins or immunohistochemistry. (A) Clear^T cleared E14.5 actin-GFP embryos, but reduced GFP fluorescence. Formamide (50%) maintained fluorescence, but failed to clear embryos. Clear^T2 cleared embryos and maintained fluorescence. (B) P0 sections (800 μm) were transparent after Clear^T2, with no volume change. (C) P11 Thy1-GFP (M-line) hippocampus section (800 μm), before and after clearing with Clear^T2; 38 images, 20 μm steps (top and middle). GFP⁺ pyramidal neurons (arrows) and dendrites (arrowheads) in CA1 region are markedly more visible after clearing; 52 images, 2.5 μm steps (bottom). GCL, granule cell layer; ML, molecular layer. (D) Sections of E14.5 optic chiasm (200 μm), immunolabeled with the radial glial marker RC2, cleared with Clear^T2; 51 images, 3 μm steps; three optical slices shown. RC2⁺ staining was observed deeper in cleared compared with pre-cleared tissue. Blue indicates Hoechst staining. (E) E11.5 whole embryos, immunolabeled with neurofilament antibody (NF) and treated with Clear^T2. NF⁺ axons were much more visible in cleared embryos (top); magnification of trigeminal axons reaching epithelial targets (bottom). (F) Section (300 μm) of postnatal mouse brain, dLGN anterogradely labeled with CTB conjugated to Alexa Fluor 594. A single neuron was filled with biocytin and immunostained with streptavidin-Alexa Fluor 647. Clearing with Clear^T2 enhanced resolution and visibility of the dendritic arbor of the neuron. Merged stack, 55 images, 2 μm steps. CTB label is in red; biocytin-filled neuron is pseudo-colored green. Scale bars: 1 mm in A,B,E; 40 μm in C; 20 μm in D,F.