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Holographic projection for multispot structured illumination microscopy
Author(s) -
WARD E.N.,
PAL R.
Publication year - 2019
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/jmi.12787
Subject(s) - holography , optics , projection (relational algebra) , computer science , computer generated holography , diffraction , microscopy , excitation , resolution (logic) , field (mathematics) , iterative reconstruction , computer vision , artificial intelligence , algorithm , physics , mathematics , quantum mechanics , pure mathematics
Summary This paper describes the design and testing of a multispot structured illumination microscopy system using computer‐generated holograms to create the required excitation patterns. Furthermore, it demonstrates the use of an adapted direct search algorithm for calculating the holograms that allows for imaging across an extended field of view. The system was tested on fixed targets and live cells yielding a two times resolution increase over conventional diffraction‐limited imaging. Lay Description We present the design and testing of a multispot structured illumination microscopy system using computer‐generated holograms to create the required excitation patterns. It demonstrates the use of an adapted direct search algorithm for calculating the holograms that allows for imaging across an extended field of view. The system was tested on fixed targets and live cells yielding a two times resolution increase over conventional diffraction‐limited imaging. The results here demonstrate that holography provides an efficient means of pattern projection for MSIM imaging. It provides a significant improvement in the efficiency of pattern projection and more importantly it allows for the testing of more diverse excitation patterns than possible with amplitude‐only projection. For example, PSF engineering using phase modulation can be easily incorporated into the calculated holograms, potentially generating subdiffraction structures in the excitation pattern. The ability to incorporate PSF engineering into SIM opens up holographic MSIM as a potential method for further increasing resolution with little or no change to the imaging system.