New microscopy platform enables imaging of neuronal activity at single-cell resolution at unprecedented tissue depth
As light travels deeper into the brain, it scatters. Three-photon microscopy, the current state of the art, reduces the resulting background fluorescence and can, in principle, image much deeper than earlier techniques—but to do that, it uses average laser power limits that cause overheating and damage living tissue.
A team of researchers at the Rockefeller University have developed a new microscopy platform called Three-photon Ultra-Deep Imaging (TrUDI) that significantly enhances the ability to image neuronal activity at single-cell resolution at unprecedented tissue depths. Traditional three-photon microscopy, while capable of imaging deeper into the brain than other techniques, has been limited by overheating and tissue damage due to average laser power limits.
The TrUDI platform addresses these limitations by reengineering the rate and energy of laser pulses, how they are delivered and scanned at the sample, all within safe limits of sample power exposure. This results in a roughly 250-fold increase in imaging efficiency, enabling researchers to image through the entire mouse cortex and record neural activity in deep brain regions, such as the hippocampus and striatum, without the need for surgical tissue removal.
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