Non-scanning motionless fluorescence three-dimensional holographic microscopy

“Holography is an attractive imaging technique as it offers the ability to view a complete three-dimensional volume from one image. However, holography is not widely applied to the field of three-dimensional fluorescence microscopic imaging, because fluorescence is incoherent and creating holograms requires a coherent interferometer system. Although scanning one beam of an interferometer pattern across the rear aperture of an objective to excite fluorescence in a specimen overcomes the coherence limitation, the mechanical scanning is complicated, which makes the image capturing slow, and the process is limited to low-numerical-aperture objectives. Here we present the first demonstration of a motionless microscopy system (FINCHSCOPE) based on Fresnel incoherent correlation holography, and its use in recording high-resolution three-dimensional fluorescent images of biological specimens. By using high-numerical-aperture objectives, a spatial light modulator, a CCD camera and some simple filters, FINCHSCOPE enables the acquisition of three-dimensional microscopic images without the need for scanning.”