Abstract
Multifocus microscopy (MFM) allows sensitive and fast three-dimensional imaging. It relies on the efficient design of diffraction phase gratings yielding homogeneous intensities in desired diffraction orders. Such performances are however guaranteed only for a specific wavelength. Here, we discuss a novel approach for designing binary phase gratings with dual color properties and improved diffraction efficiency for MFM. We simulate binary diffraction gratings with tunable phase shifts to explore its best diffraction performances. We report the design and fabrication of a binary array generator of 3 × 3 equal-intensity diffraction orders with 74% efficiency, 95% uniformity and dual color capability. The multicolor properties of this new design are highlighted by two-color MFM imaging. Finally, we discuss the basics of extending this approach to a variety of diffraction pattern designs.
Highlights
Fluorescence microscopy is a powerful tool to decipher biological processes in living cells
The Multifocus microscopy (MFM) is based on the introduction of multiple diffraction optical elements and prisms on the emission path of a wide-field microscope
For MFM, an important task is to design a phase grating diffracting the beam into a MxN array of discrete diffraction orders with equal intensity centered at the 0th order, and with the maximum attainable efficiency
Summary
In a similar manner to the 0.84 × π grating, designing a grating at a phase shift of π ± Δφ (where Δφ does not exceed 0.1 × π for these cases), two wavelength-bands appear with similar maximum efficiency performances This fact underlies that by changing Δφ value, the two bands can be tuned to the desired imaging wavelengths. It is possible to tune the two imaging wavelength bands when designing the grating without altering the efficiency and uniformity of the diffraction orders. This is not a necessary condition in the case of an odd number of diffraction orders where the zero order should remain and phase shifts other than π can result in the overall diffraction efficiency improvement
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