Signal enhanced holographic fluorescence microscopy with guide-star reconstruction

Biomed Opt Express. 2016 Mar 14;7(4):1271-83. doi: 10.1364/BOE.7.001271. eCollection 2016 Apr 1.

Abstract

We propose a signal enhanced guide-star reconstruction method for holographic fluorescence microscopy. In the late 00's, incoherent digital holography started to be vigorously studied by several groups to overcome the limitations of conventional digital holography. The basic concept of incoherent digital holography is to acquire the complex hologram from incoherent light by utilizing temporal coherency of a spatially incoherent light source. The advent of incoherent digital holography opened new possibility of holographic fluorescence microscopy (HFM), which was difficult to achieve with conventional digital holography. However there has been an important issue of low and noisy signal in HFM which slows down the system speed and degrades the imaging quality. When guide-star reconstruction is adopted, the image reconstruction gives an improved result compared to the conventional propagation reconstruction method. The guide-star reconstruction method gives higher imaging signal-to-noise ratio since the acquired complex point spread function provides optimal system-adaptive information and can restore the signal buried in the noise more efficiently. We present theoretical explanation and simulation as well as experimental results.

Keywords: (070.0070) Fourier optics and signal processing; (090.1995) Digital holography; (170.2520) Fluorescence microscopy.