Accurate 3D morphological computational model reconstruction of suspended cells imaged through stratified media by the precise depth-varying point spread function method.

Accurate three-dimensional (3D) morphological computational models of cells are important in a number of biological studies. This study proposes a precise depth-varying point spread function (PDV-PSF) method for reconstructing 3D computational models of suspended cells from two-dimensional (2D) confocal image stacks. Our approach deblurs the 2D images in horizontal plane and corrects the deformation in vertical direction to overcome the refractive index mismatch problem caused by suspended cells imaging through stratified media. Standard fluorescent polystyrene spheres and Jurkat T-lymphocytes are selected to evaluate the validity and accuracy of this PDV-PSF method. Qualitative and quantitative results demonstrate that our approach has superior performance in 3D morphological computational models reconstruction of suspended cells.

Clinical lymphocytes construction for light scattering inversion study: a three-dimensional morphology constructed method from defective confocal images.

Constructing models of cells' realistic internal and external morphology is vital for correlation between light scattering and morphology of the scattering structure. The image stack obtained from fluorescent confocal microscopy is at present used to construct the cell's three-dimensional (3-D) morphology. However, due to the poor labeling quality and unavoidable optical noise present in the image stacks, 3-D morphologies are difficult to construct and are an impediment to the statistical analyses of cell structures. We propose a method called the "area and shape constraint method (ASCM)" for constructing 3-D morphology. Blurred 3-D morphologies constructed by common methods from image stacks considered as defective and which are commonly discarded are well restored by the ASCM. Seventy-four clinical blood samples and a series of standard fluorescent spheres are selected to evaluate the validity and precision of our proposed ASCM. Both the qualitative and quantitative results obtained by ASCM indicate the good performance of the method in constructing the cell's 3-D morphology.