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   The neuroscience field strongly depends on acquiring faster images at greater depths for deep tissue in vivo imaging. Scattering media not only inhibits subsurface cell-scale imaging within tissues, such as the brain, but also limits the penetration depth of current optical imaging. Multi-photon microscopy (MPM) overcomes this problem at depths up to a few scattering mean free path lengths (e.g. 500 µm in mouse brain). Adaptive optics allows optical wavelength control needed for transformative impacts on deep tissue imaging, a goal in the neuroscience field.


   A major problem standard multi-photon microscopy faces is correcting for scattering media. Super Penetration Multi-Photon Microscopy (S-MPM), was pioneered by the Cui Lab at Howard Hughes Medical Center (currently at Purdue University) and recently reported by Boston University on focusing light through static and dynamic strongly scattering media. Developed at Boston University and commercialized by Boston Micromachines, the enabling component is a fast MEMS spatial light modulator (Kilo-S-SLM), incorporated into the test bed shown to the right. With this component, images of 1 µm diameter fluorescent beads through 280 µm thick mouse skull reached image depths of about 500  µm.


The images below depict these measurements.

Multi-Photon microscopy 

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Additional observations made:

  • 10x – 100x advance in signal enhancement with higher resolution and contrast.
  • SLM corrects for low order spherical aberrations as well as higher order scattering effects.
  • The optimized SLM phase improves imaging over a field of view of 10–20 µm for samples tested to date.





Optical Window ~20 µm

PMT Image (Reflection)

Camera image (Transmission before optimization)

multi photon microscopy with boston micromachines adaptive optics
multi photon microscopy with boston micromachines adaptive optics
multi photon microscopy with boston micromachines adaptive optics
Multi Photon Microscopy with Deformable Mirror Adaptive Optics
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Learn more about the Boston Micromachines Deformable Mirror model used for both The Howard Hughes Medical Institute and Boston University:​

​Imaging fluorescent beads (1 µm diameter) through mouse skull (280 µm thick) using a glass cover slip (150 µm thick).

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