ADAPT-3D™ Tissue Clearing Kit

1. Decolorization Buffer (B671)
2. Partial Delipidation Buffer (Component A) (B672)
3. Partial Delipidation Buffer (Component B) (B676) Note: Component B is corrosive to most plastic types; use glass tubes.
4. 1X ADAPT-3D Staining Buffer (B673)
5. Refractive Index Matching (Clearing Solution) (B675)
6. Wash Buffer (5X) (B674)

Please note that required buffers and materials not included in the kit are: Cell lines, organoids, tissue or animal organs, slips, coverslips, containers, scintillation vials, primary and secondary antibodies, Decalcification Buffer (Leinco SKU: B670), and Fixative Solution – 4% paraformaldehyde (PFA) in PBS with 10% sucrose at pH 9.0.

ADAPT-3D (Accelerated Deep Adaptable Processing of Tissue for 3-Dimensional Imaging), has an adaptable and versatile nature for use with a wide range of tissues and model systems. This technology is designed for microscopy to prepare biological tissues for 3D fluorescence imaging. The ADAPT-3D method involves a protocol that serves as a guide with recommended incubation times for fixation, decolorization, delipidation, and refractive index matching (RIM). The ADAPT-3D method was developed with inspiration from advances in aqueous-based clearing methods to achieve rapid, aqueous 3D imaging.

USES
The ADAPT-3D Tissue Clearing Kit is used to prepare fixed tissue samples for 3-dimensional fluorescence tissue imaging. This allows for improved light penetration in 3D tissues, enhancing the detection of fluorescent signals deeper within the samples. The method is broadly applicable across species and tissue types. Specific uses highlighted in the sources include:
• Imaging human intestinal tissue after fixation, decolorization, delipidation, and antibody staining.
• Imaging intact mouse skull and brain to detect specialized channels between the skull and underlying tissue without meningeal tearing.
• Whole-organ imaging such as mouse brain, spleen, lung, and liver with retention of endogenous fluorescent reporters.
• Light-sheet microscopy of cleared whole mouse brains with strong retention of fluorescent intensities of endogenous reporter proteins like eYFP and tdTomato.
• Imaging bone with adjacent soft tissues like the skull-brain interface after decalcification.
• Deep immunolabeling and detection of antigens such as tight junctional proteins, smooth muscle actin, S100A9, CD163, and IBA1 in various tissues like leptomeninges and intestine.
• Compatible with multiple imaging modalities including widefield, confocal, and light-sheet microscopy.