Superresolved Multiphoton Microscopy with Spatial Frequency-Modulated Imaging

Dean Levi, Jeffrey Field, Keith Wernsing, Scott Domingue, Alyssa Allende Motz, Keith DeLuca, Jennifer DeLuca, Michael Young, Jeff Squier, Randy Bartels

Research output: Contribution to journalArticlepeer-review

64 Scopus Citations

Abstract

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..eta.. below the diffraction limit, where ..eta.. 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.
Original languageAmerican English
Pages (from-to)6605-6610
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number24
DOIs
StatePublished - 2016

NREL Publication Number

  • NREL/JA-5J00-66628

Keywords

  • harmonic generation
  • multiphoton microscopy
  • superresolution

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