Single shot, three-dimensional fluorescence microscopy with a spatially rotating point spread function

Fluorescence microscopy three-dimensional depth variant point spread function interpolation using Zernike moments

In three-dimensional fluorescence microscopy the point spread function (PSF) changes with depth, inducing errors in the restored images when these variations are neglected during the deconvolution of thick specimens. Some deconvolution algorithms have been developed to take the depth variations of the PSF into consideration. For these algorithms, the accuracy of the estimated structures depends...

Enhanced three-dimensional deconvolution microscopy using a measured depth-varying point-spread function.

We present a technique to systematically measure the change in the blurring function of an optical microscope with distance between the source and the coverglass (the depth) and demonstrate its utility in three-dimensional (3D) deconvolution. By controlling the axial positions of the microscope stage and an optically trapped bead independently, we can record the 3D blurring function at differen...

Three-dimensional, single-molecule fluorescence imaging beyond the diffraction limit by using a double-helix point spread function.

We demonstrate single-molecule fluorescence imaging beyond the optical diffraction limit in 3 dimensions with a wide-field microscope that exhibits a double-helix point spread function (DH-PSF). The DH-PSF design features high and uniform Fisher information and has 2 dominant lobes in the image plane whose angular orientation rotates with the axial (z) position of the emitter. Single fluorescen...

Three-dimensional holographic fluorescence microscopy.

Most commonly used methods for three-dimensional (3D) fluorescence microscopy make use of sectioning techniques that require that the object be physically scanned in a series of two-dimensional (2D) sections along the z axis. The main drawback in these approaches is the need for these sequential 2D scans. An alternative approach to fluorescence imaging in three dimensions has been developed tha...

Multi-kernel deconvolution applied to confocal fluorescence microscopy with engineered point spread function