Improvement of axial resolution and contrast in temporally focused widefield two-photon microscopy with structured light illumination
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Updated: May 9, 2026

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Published on: May 30, 2016
Heejin Choi1, Elijah Y S Yew, Bertan Hallacoglu
1Department of Mechanical Engineering Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Abstract:
Although temporally focused wide-field two-photon microscopy (TFM) can perform depth resolved wide field imaging, it cannot avoid the image degradation due to scattering of excitation and emission photons when imaging in a turbid medium. Further, its axial resolution is inferior to standard point-scanning two-photon microscopy. We implemented a structured light illumination for TFM and have shown that it can effectively reject the out-of-focus scattered emission photons improving image contrast. Further, the depth resolution of the improved system is dictated by the spatial frequency of the structure light with the potential of attaining depth resolution better than point-scanning two-photon microscopy.
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