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Adaptive Optics via Pupil Image Segmentation

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Imaging Instrumentation
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Adaptive Optics via Pupil Image Segmentation

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For High Resolution Imaging in Biological Tissues

Microscope Adaptive Optics by rear pupil masking, adaptive optics for high-resolution imaging in biological tissue.

This technology advances light microscopy's resolution and depth limit using an innovative and effective approach: adaptive optics (AO). The inherent heterogeneity of biological specimens limits depth and resolution in life science imaging. In addition, these specimens give rise to optical distortions, known as aberrations that lead to signal loss, image fidelity, and resolution. AO has been used in telescopes to address the problem of optical aberration of light beams as they propagate through our atmosphere, but it has been applied to microscopy in only a limited manner due to technical challenges. This new technology is an image-based AO method that is simpler to implement than other AO methods and suitable for microscopy. This innovation comprises the use of an SLM in the optical path of a microscope and proprietary algorithms. It attains near-diffraction-limited performance from a variety of biological samples with only minor modifications of a light microscope.

The value of the technology has been demonstrated in two-photon microcopy, where it attained near-diffraction-limited performance in varied biological and non-biological samples exhibiting aberrations large or small and smoothly varying or abruptly changing. Work in fixed mouse cortical slices illustrates the capability to improve signal and resolution to depths of 500m. The technology applies to other imaging modalities, including confocal microscopy and widefield microscopy.


  • Improved signal and resolution to depths of 500m
  • Near diffraction-limited resolution
  • Requires only minor modification of the microscope
  • It can be used with two-photon, confocal, and wide-field microscopy


  • Basic life science research
  • Imaging of living or thick biological specimens
  • In vivo calcium imaging
  • Two-photon microscopy
  • CARS, second harmonic generation, and third harmonic generation imaging

Patent Status:

Issued U.S. Patent 8,730,573


Free for Non-Profit Research and available for Commercial License. Contact our Innovation Management Department at

For inquiries, please reference:

Janelia 2009-004

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Michael Perham
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