Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging.

The ultimate aim of fluorescence microscopy is to achieve high-resolution imaging of increasingly larger biological samples. Extended depth of field presents a potential solution to accelerate imaging of large samples when compression of information along the optical axis is not detrimental to the interpretation of images. We have implemented an extended depth of field (EDF) approach in a random illumination microscope (RIM).

Super-resolved live-cell imaging using random illumination microscopy.

Current super-resolution microscopy (SRM) methods suffer from an intrinsic complexity that might curtail their routine use in cell biology. We describe here random illumination microscopy (RIM) for live-cell imaging at super-resolutions matching that of 3D structured illumination microscopy, in a robust fashion. Based on speckled illumination and statistical image reconstruction, easy to implement and user-friendly, RIM is unaffected by optical aberrations on the excitation side, linear to brightness, and compatible with multicolor live-cell imaging over extended periods of time.

Performance model of depth from defocus with an unconventional camera.

In this paper, we present a generic performance model able to evaluate the accuracy of depth estimation using depth from defocus (DFD). This model only requires the sensor point spread function at a given depth to evaluate the theoretical accuracy of depth estimation. Hence, it can be used for any (un)conventional system, using either one or several images.