Noninvasive megapixel fluorescence microscopy through scattering layers by a virtual incoherent reflection matrix

Area of Science:

• Biomedical Optics
• Microscopy
• Photonics

Background:

• Optical imaging in complex samples is hindered by light scattering.
• Existing methods for scattering compensation in fluorescence imaging have limitations, including target sparsity or complex requirements.
• Reflection matrix techniques have shown promise for coherent imaging but are not readily applicable to incoherent fluorescence.

Purpose of the Study:

• To adapt reflection matrix techniques for scattering compensation in incoherent fluorescence imaging.
• To develop a method for reconstructing high-resolution fluorescence images from limited measurements in scattering media.
• To overcome the limitations of current scattering compensation approaches in fluorescence microscopy.

Main Methods:

• Developed a virtual fluorescence-based reflection matrix from conventional wide-field microscope images.
• Acquired images under unknown, random illumination patterns.
• Applied an adapted matrix-based scattering compensation algorithm to the constructed matrix.

Main Results:

• Successfully reconstructed megapixel-scale images from fewer than 150 acquired frames.
• Demonstrated effective scattering compensation without spatial light modulators or intensive computation.
• Validated the approach experimentally in complex scattering samples.

Conclusions:

• The presented approach effectively compensates for scattering in incoherent fluorescence imaging.
• This method offers a computationally efficient and practical solution for high-resolution imaging through scattering media.
• The technique has broad implications for various fields requiring deep or through-sample optical imaging.