SHIELD preserves protein fluorescence and antigenicity, transcripts and tissue architecture under a wide range of harsh conditions. SHIELD was applied to interrogate system-level wiring, synaptic architecture, and molecular features of virally labeled neurons and their targets in mouse at single-cell resolution. Rapid three dimensional phenotyping of core needle biopsies and human bran cells was also demonstrated. SHIELD enables rapid, multiscale, integrated molecular phenotyping of both animal and clinical tissues.
CLARITY enables the transformation of intact tissue into a nanoporous hydrogel-hybridized form (crosslinked to a three-dimensional network of hydrophilic polymers) that is fully assembled but optically transparent and macromolecule-permeable. Using a mouse brain, intact-tissue imaging of long-range projections, local circuit wiring, cellular relationships, subcellular structures, protein complexes, nucleic acids and neurotransmitters are shown.
SWITCH introduces a simple method that enables proteomic imaging for scalable, integrated, high-dimensional phenotyping of both animal tissues and human clinical samples. The technology uniformly secures tissue architecture, native biomolecules, and antigenicity across an entire system by synchronizing the tissue preservation reaction. The framework permits multiple rounds (>20) of relabeling while synchronizing labeling reactions to improve probe penetration depth and uniformity of staining.
Stochastic Eletrotransport introduces a novel electrokinetic method for rapid nondestructive processing of porous samples. This method uses a rotational electric field to selectively disperse highly electromobile molecules throughout a porous sample without displacing the low-electromobility molecules that constitute the sample. Computational models show stochastic electrotransport can rapidly disperse electromobile molecules in a porous medium. This method enables completely clearing mouse organs within 1–3 days and staining them with nuclear dyes, proteins, and antibodies within 1 day. The results demonstrate the potential of stochastic electrotransport to process large and dense tissue samples that were previously infeasible in time when relying on diffusion.
Magnified Analysis of Proteome (MAP) linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. The expanded tissue tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. Off-the-shelf antibodies can be used for multiple rounds of immunolabeling and imaging of a tissue’s magnified proteome.
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