Abstract
To investigate the cellular structure, biomedical researchers often obtain three-dimensional images by combining two-dimensional images taken along the z axis. However, these images are blurry in all directions due to diffraction limitations. This blur becomes more severe when focusing further inside the specimen as photons in deeper focus must traverse a longer distance within the specimen. This type of blur is called depth-variance. Moreover, due to lens imperfection, the blur has asymmetric shape. Most deconvolution solutions for removing blur assume depth-invariant or x-y symmetric blur, and presently, there is no open-source for depth-variant asymmetric deconvolution. In addition, existing datasets for deconvolution microscopy also assume invariant or x-y symmetric blur, which are insufficient to reflect actual imaging conditions. DVDeconv, that is a set of MATLAB functions with a user-friendly graphical interface, has been developed to address depth-variant asymmetric blur. DVDeconv includes dataset, depth-variant asymmetric point spread function generator, and deconvolution algorithms. Experimental results using DVDeconv reveal that depth-variant asymmetric deconvolution using DVDeconv removes blurs accurately. Furthermore, the dataset in DVDeconv constructed can be used to evaluate the performance of microscopy deconvolution to be developed in the future.
Highlights
One of the most basic imaging techniques in biomedical research is wide-field fluorescence microscopy
In wide-field fluorescence microscopy, a dye-labeled specimen is illuminated with light that matches the excitation spectrum of the dye, and emitted light is captured by a camera [1]
Deconvolution Results Peak signal-to-noise ratio (PSNR), signal-to-noise ratio (SNR), standard deviation of peaks, relative contrast, memory, and processing time were used to measure the performance of the DVDeconv algorithms on synthetic data
Summary
One of the most basic imaging techniques in biomedical research is wide-field fluorescence microscopy. In wide-field fluorescence microscopy, a dye-labeled specimen is illuminated with light that matches the excitation spectrum of the dye, and emitted light is captured by a camera [1]. Researchers can obtain a three-dimensional (3D) specimen image by taking two-dimensional (2D) fluorescence microscopy images along the z axis [2]. A disadvantage of obtaining 3D specimen images using this technique is that the image captured is blurry because of diffraction and lens aberrations. The PSF in the lateral (x-y) plane contains the Airy disk, while the PSF in the z (x-z or y-z) plane has an hourglass shape. Some refractive index changes or lens aberration are present, and the PSF shape is distorted as an asymmetric Airy disk and hourglass shape
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