Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 113, Issue 24, Pages 6605-6610Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1602811113
Keywords
superresolution; harmonic generation; multiphoton microscopy
Categories
Funding
- W. M. Keck Foundation
- Institute for Genome Architecture and Function at Colorado State University
- National Institute of Mental Health [2R42MH102201]
- National Institute of Biomedical Imaging and Bioengineering under Bioengineering Research Partnership [EB-00382]
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Superresolved far-field microscopy has emerged as a powerful tool for investigating the structure of objects with resolution well below the diffraction limit of light. Nearly all superresolution imaging techniques reported to date rely on real energy states of fluorescent molecules to circumvent the diffraction limit, preventing superresolved imaging with contrast mechanisms that occur via virtual energy states, including harmonic generation (HG). We report a superresolution technique based on spatial frequency-modulated imaging (SPIFI) that permits superresolved nonlinear microscopy with any contrast mechanism and with single-pixel detection. We show multimodal superresolved images with two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) from biological and inorganic media. Multiphoton SPIFI (MP-SPIFI) provides spatial resolution up to 2. below the diffraction limit, where. is the highest power of the nonlinear intensity response. MP-SPIFI can be used to provide enhanced resolution in optically thin media and may provide a solution for superresolved imaging deep in scattering media.
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