Confocal rescan structured illumination microscopy for real-time deep tissue imaging with superresolution

Abstract

Structured illumination microscopy (SIM) is an established optical superresolution imaging technique. However, conventional SIM based on wide-field image acquisition is generally limited to visualizing thin cellular samples. We propose combining one-dimensional image rescan and structured illumination in the orthogonal direction to achieve superresolution without the need to rotate the illumination pattern. The image acquisition speed is consequently improved threefold, which is also beneficial for minimizing photobleaching and phototoxicity. Optical sectioning in thick biological tissue is enhanced by including a confocal slit in the system to significantly suppress the out-of-focus background and the associated noise. With all the technical improvements, our method captures threedimensional superresolved image stacks of neuronal structures in mouse brain tissue samples for a depth range of more than 200 μm.

Principle and Recent Development in Photonic Time-Stretch Imaging

“…The recent development of PTS imaging systems are mainly in four categories: first, PTS imaging systems combined with shorter wavelength bands for better imaging resolutions and rich applications [68][69][70][71][72][73]; second, PTS imaging systems with faster speed [21,22,[26][27][28][29]57,58,74,79]; third, PTS imaging systems combined with data compression [11][12][13][14][15][16][17][18][75][76][77][78]; and the last, PTS imaging systems combined with deep learning for target classification [23][24][25]32,66].…”

“…Recent developments in optical imaging have paved the way for diversified applications in science and industry, especially around medicine, biochemistry, and biology . For example, noninvasive diagnosis of biomedical tissue using optical coherence tomography (OCT) [1][2][3][4][5][6][7][8][9][10], ultrafast optical imaging based on photonic time-stretch structure , data compressive optical imaging systems [10][11][12][13][14][15][16][17][18][19][20][21][22], blood screening and abnormal cell screening using contrast microscopy [23][24][25][26][27], these developments of optical imaging are with the support of digital technologies [32][33][34][35][36][37][38][39] and the growing industry.…”