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Second harmonic generation (SHG) microscopy is a valuable tool for optical microscopy. SHG microscopy is normally performed as a point scanning imaging method, which lacks phase information and is limited in spatial resolution by the spatial frequency support of the illumination optics. In addition, aberrations in the illumination are difficult to remove. We propose and demonstrate SHG holographic synthetic aperture holographic imaging in both the forward (transmission) and backward (epi) imaging geometries. By taking a set of holograms with varying incident angle plane wave illumination, the spatial frequency support is increased and the input and output pupil phase aberrations are estimated and corrected – producing diffraction limited SHG imaging that combines the spatial frequency support of the input and output optics. The phase correction algorithm is computationally efficient and robust and can be applied to any set of measured field imaging data.
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Synthetic spatial aperture holographic third harmonic generation microscopy
Third harmonic generation (THG) provides a valuable, label-free approach to imaging biological systems. To date, THG microscopy has been performed using point-scanning methods that rely on intensity measurements lacking phase information of the complex field. We report the first demonstration, to the best of our knowledge, of THG holographic microscopy and the reconstruction of the complex THG signal field with spatial synthetic aperture imaging. Phase distortions arising from measurement-to-measurement fluctuations and imaging components cause optical aberrations in the reconstructed THG field. We have developed an aberration-correction algorithm that estimates and corrects these phase distortions to reconstruct the spatial synthetic aperture THG field without optical aberrations.
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- Award ID(s):
- 2006416
- PAR ID:
- 10507328
- Publisher / Repository:
- Optical Society of America
- Date Published:
- Journal Name:
- Optica
- Volume:
- 11
- Issue:
- 5
- ISSN:
- 2334-2536
- Format(s):
- Medium: X Size: Article No. 693
- Size(s):
- Article No. 693
- Sponsoring Org:
- National Science Foundation
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