Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues

T Ku, J Swaney, JY Park, A Albanese, E Murray… - Nature …, 2016 - nature.com
T Ku, J Swaney, JY Park, A Albanese, E Murray, JH Cho, YG Park, V Mangena, J Chen
Nature biotechnology, 2016nature.com
The biology of multicellular organisms is coordinated across multiple size scales, from the
subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell
populations. Here we introduce a method for super-resolution imaging of the multiscale
organization of intact tissues. The method, called magnified analysis of the proteome (MAP),
linearly expands entire organs fourfold while preserving their overall architecture and three-
dimensional proteome organization. MAP is based on the observation that preventing …
Abstract
The biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.
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