Abstract
Imaging three-dimensional structures represents a major challenge for conventional microscopies. Here we describe a Spatial Light Modulator (SLM) microscope that can simultaneously address and image multiple targets in three dimensions. A wavefront coding element and computational image processing enables extended depth-of-field imaging. High-resolution, multi-site three-dimensional targeting and sensing is demonstrated in both transparent and scattering media over a depth range of 300-1,000 microns.
Highlights
The natural world is fundamentally three dimensional
The relative intensity of the CP-PSF compared to the performance of a conventional microscope shows a significant increase in the localization of signal at large defocus if sufficient α is applied, as shown in Fig. 2(C) – motivating our use of extended Depth of Field (DOF) techniques
It is the prior knowledge provided by the user-controlled illumination device that is integral to providing context to the images acquired by the extended Depth of Field microscope
Summary
The natural world is fundamentally three dimensional This simple fact has long presented difficulties for conventional high numerical aperture (NA) imaging systems, where the axial depth of focus scales as NA 2. As a result, when using high NA optics to image volumes, it has been necessary to acquire data sequentially, which limits the speed of imaging and the ability to image multiple sites across the volume with true simultaneity. This is especially problematic when one wants to image in complex systems that exhibit distinct temporal dynamics at each point in space, such as imaging the activity of neurons in complete neural circuits - if possible simultaneously [1]
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