Optimal resolution in Fresnel Incoherent Correlation Holographic fluorescence microscopy

Gary Brooker, Nisan Siegel, Victor Wang, Joseph Rosen

Research output: Contribution to journalArticlepeer-review

113 Scopus citations

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.

Original languageEnglish
Pages (from-to)5047-5062
Number of pages16
JournalOptics Express
Volume19
Issue number6
DOIs
StatePublished - 14 Mar 2011

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics

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