The Smith laboratory is developing a new, ultrahigh-resolution immunoimaging method called Array Tomography. By permitting three-dimensional visualization of biomolecular architectures with unprecedented clarity, this new method fills important gaps in previous abilities to analyze cells and tissues and to diagnose human illness. Array tomography is proteomic in scope, being capable of imaging an almost unlimited number of distinct cellular molecules in individual specimens, and improves greatly upon existing immunofluorescence microscopy techniques in quantitative reliability, spatial resolution, sensitivity and specificity. Array tomography complements dynamic in vivo imaging methods, including those based on the use of expressed fluorescent proteins, by providing efficient means for retrospective immunoanalysis of dynamically imaged specimens. In addition, array tomography will enable exploration of the most intricate tissue architectures, such as the brain’s synaptic circuitry, that have proved largely intractable to existing methods. To learn more about Array Tomography visit the Stanford Array Tomography Resource (SATR) website.

Motion Rendering from Array Tomograms

These thumbnail movies were rendered from array tomograms of mature mouse cortex and illustrate the extremey high volumetric resolution with which specific cytoskeletal, synaptic and ion channel antigens and fluorescent proteins can be imaged. (Ion channel antibodies provided by Prof. James Trimmer, UC Davis).

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Presynaptic boutons (red - synapsin I) and dendritic shafts (green - tubulin) in mouse hippocampal cortex

Presynaptic boutons (red - synapsin I) and genetically encoded fluorescent protein (green) in "Line M" GFP transgenic mouse cerebral cortex

A slab of mouse cerebral cortex showing punctate distribution of potassium channel subunits (KV2.1 - green) in relation to presynaptic boutons (synapsin I - red) and DNA (DAPI - blue).

A higher magnification rendering from the same array tomograms as that at left.

 

Rendering from a tubulin-stained array tomogram of mouse cortex. The core microtubule bundles that delineate the trajectories of all mature axons and dendrites are evident here.