PMID-21280920 Optically sectioned in vivo imaging with speckle illumination HiLo microscopy
- Ah, brilliant! Illuminate a sample with a speckle pattern from a laser, and use this to optically section the data -- the contrast of the speckle pattern shows how in focus the sample is.
- Hanece, the contrast indicates the in-focus vs out-of-focus ratio in a region.
- The speckle statistics are invariant even in a scattering media, as scattering only further randomizes an already random laser phase front. (Within some limits.)
- HiLo microscopy involves illuminating with a speckle pattern, then illuminating with standard uniform illumination, resulting in a diffraction-limited optically sectioned image. PMID-18709098
- Algorithm is :
- Take the speckle image and subtract the uniform image
- Bandpass
- Measure the standard deviation of the to get a weighting function
- Debias this estimate based on sensor..
- Generate low-passed image from the weighted uniform image, , and high-pass from the difference
- Resultand image is a weighted sum of highpassed and lowpassed images.
- Looks about as good as confocal.
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- Cited by...
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