Abstract
Fresnel Incoherent Correlation Holography (FINCH) enables holograms and 3D images to be created from incoherent light with just a camera and spatial light modulator (SLM). We previously described its application to microscopic incoherent fluorescence wherein one complex hologram contains all the 3D information in the microscope field, obviating the need for scanning or serial sectioning. We now report experiments which have led to the optimal optical, electro-optic, and computational conditions necessary to produce holograms which yield high quality 3D images from fluorescent microscopic specimens. An important improvement from our previous FINCH configurations capitalizes on the polarization sensitivity of the SLM so that the same SLM pixels which create the spherical wave simulating the microscope tube lens, also pass the plane waves from the infinity corrected microscope objective, so that interference between the two wave types at the camera creates a hologram. This advance dramatically improves the resolution of the FINCH system. Results from imaging a fluorescent USAF pattern and a pollen grain slide reveal resolution which approaches the Rayleigh limit by this simple method for 3D fluorescent microscopic imaging.
Highlights
Incoherent [1,2] and partially coherent [3] digital holographic microscopies have recently become fields of much interest because of the ability of microscopes based on these principles to image three dimensional (3D) incoherent objects
The holographic method used in this study is based upon the recently invented system of a single-channel incoherent interferometer employed for generating digital Fresnel holograms [6,7]
For every source point the spatial light modulator (SLM) is used as a diffractive beam splitter in an incoherent interferometer, so that each spherical beam, originating from each object point, is split into two spherical beams with two different curve radii
Summary
Incoherent [1,2] and partially coherent [3] digital holographic microscopies have recently become fields of much interest because of the ability of microscopes based on these principles to image three dimensional (3D) incoherent objects. A lensless version of a partially coherent [3], digital holographic microscope has been installed on-chip in a very compact configuration. The holographic method used in this study is based upon the recently invented system of a single-channel incoherent interferometer employed for generating digital Fresnel holograms [6,7]. In this non-scanning holographic technique, incoherent light is reflected or emitted from a 3D object, propagates through a spatial light modulator (SLM), and is recorded by a digital camera. Three holograms are recorded sequentially, each for a different phase factor of the SLM, and are superposed during data processing to produce a complex-valued Fresnel hologram free of the twin image and bias term
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