Abstract
We report a widefield super-resolution (SR) fluorescence microscopy technique termed total internal reflection fluorescence pattern-illuminated Fourier ptychographic microscopy (TIRF-piFPM). It employs pattern-illuminated Fourier ptychography (piFP) under the total internal reflection fluorescence (TIRF) mode, providing a lateral resolution of ∼100 nm, reducing the background level, and correcting unknown optical aberrations. Like the total internal reflection fluorescence structure illuminated microscopy (TIRF-SIM), the illumination field of TIRF-piFPM is modulated by sinusoidal patterns generated by evanescent wave interference. It differs from TIRF-SIM in that TIRF-piFPM reconstructs the SR image by FP iteration. To demonstrate the performance of TIRF-piFPM, we compare it with TIRF-SIM by conducting simulations and experiments and prove that TIRF-piFPM can provide a better result than TIRF-SIM in terms of its robustness to noise and aberration correction ability. In addition, dynamic changes of mitochondria in U2OS cells are captured with a temporal resolution of 2 s, demonstrating its live-cell imaging capability. The addvantages may enable TIRF-piFPM to serve as an alternative to SR fluorescence microscopes in the TIRF family, with promising applications in biological and biomedical imaging.
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