Abstract
Fluorescence microscopy is widely used in biological imaging, however scattering from tissues strongly limits its applicability to a shallow depth. In this work we adapt a methodology inspired from stellar speckle interferometry, and exploit the optical memory effect to enable fluorescence microscopy through a turbid layer. We demonstrate efficient reconstruction of micrometer-size fluorescent objects behind a scattering medium in epi-microscopy, and study the specificities of this imaging modality (magnification, field of view, resolution) as compared to traditional microscopy. Using a modified phase retrieval algorithm to reconstruct fluorescent objects from speckle images, we demonstrate robust reconstructions even in relatively low signal to noise conditions. This modality is particularly appropriate for imaging in biological media, which are known to exhibit relatively large optical memory ranges compatible with tens of micrometers size field of views, and large spectral bandwidths compatible with emission fluorescence spectra of tens of nanometers widths.
Highlights
Fluorescence microscopy is widely used in biological imaging, scattering from tissues strongly limits its applicability to a shallow depth
In this work we show that micrometer-scale fluorescent objects hidden behind a scattering layer can be imaged non-invasively in a regular inverted wide-field microscope with single shot acquisition
We have shown how to reconstruct micrometer sized fluorescent objects through scattering media from a single intensity image of scattered light in a regular wide-field inverted microscope
Summary
Fluorescence microscopy is widely used in biological imaging, scattering from tissues strongly limits its applicability to a shallow depth. We demonstrate efficient reconstruction of micrometer-size fluorescent objects behind a scattering medium in epimicroscopy, and study the specificities of this imaging modality (magnification, field of view, resolution) as compared to traditional microscopy. Using a modified phase retrieval algorithm to reconstruct fluorescent objects from speckle images, we demonstrate robust reconstructions even in relatively low signal to noise conditions. This modality is appropriate for imaging in biological media, which are known to exhibit relatively large optical memory ranges compatible with tens of micrometers size field of views, and large spectral bandwidths compatible with emission fluorescence spectra of tens of nanometers widths
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