AI Article Synopsis

  • Two-photon fluorescence microscopy (2PM) is limited in depth when imaging mouse brains due to tissue scattering, typically visualizing only the cortical layer.
  • Three-photon fluorescence microscopy (3PM) offers a non-invasive technique to image deeper subcortical structures in intact mouse brains, using a longer excitation wavelength of 1,700 nm.
  • This advancement allows for detailed imaging of vascular structures and RFP-labeled neurons in the hippocampus, enabling more in-depth biological investigations.

Article Abstract

Two-photon fluorescence microscopy (2PM) enables scientists in various fields including neuroscience, embryology, and oncology to visualize and tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of 2PM within the mouse brain to the cortical layer, and imaging subcortical structures currently requires the removal of overlying brain tissue or the insertion of optical probes. Here we demonstrate non-invasive, high resolution, imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy (3PM) at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein (RFP)-labeled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher order nonlinear excitation overcomes the limitations of 2PM, enabling biological investigations to take place at greater depth within tissue.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3864872PMC
http://dx.doi.org/10.1038/nphoton.2012.336DOI Listing

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