Fluorescence incoherent color holography

Abstract

We present a new imaging method to record multicolor digital holograms from objects emitting fluorescent light. The fluorescent light specific to the emission wavelength of various fluorescent dyes after excitation of three dimensional (3D) objects is recorded on a digital monochrome camera after reflection from a diffractive optical element (DOE). For each wavelength of fluorescent emission, the camera sequentially records three holograms reflected from the DOE, each with a different phase factor of the DOE's function. The three holograms are superposed in a computer to create a complex valued Fresnel hologram of each fluorescent emission. The holograms for each fluorescent color are further combined in a computer to produce a multicolored fluorescence hologram and 3D color image.

Review of three-dimensional holographic imaging by Fresnel incoherent correlation holograms

“…This review concentrates on the technique of incoherent digital holography based on single-channel incoherent interferometers, which we have been involved in their development recently [16][17][18][19] . The type of hologram discussed here is the digital Fresnel hologram, which means that a hologram of a single point has the form of the well-known FZP.…”

“…The discussed incoherent digital hologram is dubbed Fresnel incoherent correlation hologram (FINCH) [16][17][18] . The FINCH is actually based on a single channel onaxis incoherent interferometer.…”

“…Various aspects of the FINCH have been described in Refs. [16][17][18][19], including FINCH of reflected white light 16 , FINCH of fluorescence objects 17 , a FINCH-based holographic fluorescence microscope 18 and finally a hologram recorder in a mode of a synthetic aperture 19 . We briefly review these works in the current section.…”

“…Besides a constant term, the hologram expression contains two terms of correlation between an object and a quadratic phase, Z-dependent, function. In order to remain with a single correlation term out of the three terms, we follow the usual procedure of on-axis digital holography 14,[16][17][18][19] . Three holograms of the same object are recorded each of which with a different phase constant.…”

Improving Phase Contrast of Digital Holographic Microscope using Spatial Light Modulator

“…In this study, we demonstrate a new method for obtaining phase shifting with a higher phase contrast in the reconstructed 3D hologram using a spatial light modulator (SLM) and an in-line DHM [16]. The key component of this method is an SLM that can display phase information.…”