Abstract
Three-dimensional imaging in biological samples usually suffers from performance degradation caused by optical inhomogeneities. Here we proposed an approach to adaptive optics in fluorescence microscopy where the aberrations are measured by self-interference holographic recording and then corrected by a post-processing optimization procedure. In our approach, only one complex-value hologram is sufficient to measure and then correct the aberrations, which results in fast acquisition speed, lower exposure time, and the ability to image in three-dimensions without the need to scan the sample or any other element in the system. We show proof-of-principle experiments on a tissue phantom containing fluorescence particles. Furthermore, we present three-dimensional reconstructions of actin-labeled MCF7 breast cancer cells, showing improved resolution after the correction of aberrations. Both experiments demonstrate the validity of our method and show the great potential of non-scanning adaptive three-dimensional microscopy in imaging biological samples with improved resolution and signal-to-noise ratio.
Highlights
Quantitative imaging of integral refractive index and thickness of living cells by digital holographic microscopy (DHM) has benefits the study of cellular dynamics [1,2]
While the applications of DHM are restricted to the coherent imaging system, on the other hand, over past several years digital holography of spatial incoherent fluorescent samples has emerged as an attractive scanning-free three-dimensional (3D) imaging technique
On the charge coupled device (CCD) plane the interference pattern has the form of Ii, Ii where fSLM is the focal length of the positive lens generated by the spatial light modulator (SLM), θi is the i-th phaseshift value and φa is the phase of the introduced optical aberrations
Summary
Quantitative imaging of integral refractive index and thickness of living cells by digital holographic microscopy (DHM) has benefits the study of cellular dynamics [1,2]. Indirect sensor-less AO methods have been proposed and developed to overcome the limitations of SHWS [15,16,17,18,19] In these methods, tens of pictures of the same scene are required to detect the aberrations, which can lead to long total exposure time in cases where aberrations are substantial. We propose a FINCH-based 3D fluorescence microscope with guide-star-free anisotropic aberration correction, in which the blurred, complex-value images in FINCH are compensated and improved by a digital optimization procedure. In this procedure, an image quality metric is optimized by changing a 2D phase filter in the Fourier domain. We will discuss the advantages, limitations and potential applications of our method
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