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Image Scanning Microscopy (ISM) super-resolution microscopy has gained momentum for its almost instantaneous improved resolution capabilities. Multiphoton ISM utilizes the shorter emission wavelength and confocal acquisition by exploiting each pixel on the camera as a pinhole, and numerical enhancement to achieve sub-diffraction-limit resolution. We present the use of a multiplexed approach for signal acquisition using a regular EMCCD camera. A spatiotemporal modulation scheme is employed to direct the ultrafast laser pulses to select foci within a field-of-view. Combined with a novel image acquisition method, we show that only 49 images are required to achieve a resolution of 100 nm.
Kayvan Forouhesh Tehrani andStephen A. Boppart
"Label-free multi-photon super-resolution microscopy by multiplexed single point spread function image acquisition", Proc. SPIE PC11966, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX, PC119660B (2 March 2022); https://doi.org/10.1117/12.2607832
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Kayvan Forouhesh Tehrani, Stephen A. Boppart, "Label-free multi-photon super-resolution microscopy by multiplexed single point spread function image acquisition," Proc. SPIE PC11966, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIX, PC119660B (2 March 2022); https://doi.org/10.1117/12.2607832