Researchers have devised a new computational de-scattering approach termed de-scattering with excitation patterning in temporal focusing microscopy (DEEP-TFM). They employed two-photon patterned excitation with wide-field temporal focusing, but the signal was detected using a wide-field imaging detector.
They created a modified temporal focusing microscope that uses a digital micromirror device to transmit arbitrary excitation patterns onto the focal plane. A camera detects the emission light from the modulated excitation. Excitation patterns maintain their fidelity despite traversing via scattering surfaces because of NIR wavelengths.
Scattering at or near the surface has a minor impact on TFM images; nevertheless, it decreases the images’ high-frequency information as the imaging depth increases. DEEP-TFM combines excitation pattern information with captured images to reconstruct a de-scattered image computationally. In wide-field imaging, the number of images required to de-scatter a single FOV depends only on the imaging depth.