Image adaptive point-spread function estimation and deconvolution for in vivo confocal microscopy

Visualizing deep inside the tissue of a thick biological sample often poses severe constraints on image conditions. Standard restoration techniques (denoising and deconvolution) can then be very useful, allowing one to increase the signal-to-noise ratio and the resolution of the images. In this paper, we consider the problem of obtaining a good determination of the point-spread function (PSF) of a confocal microscope, a prerequisite for applying deconvolution to three-dimensional image stacks acquired with this system. Because of scattering and optical distortion induced by the sample, the PSF has to be acquired anew for each experiment. To tackle this problem, we used a screening approach to estimate the PSF adaptively and automatically from the images. Small PSF-like structures were detected in the images, and a theoretical PSF model reshaped to match the geometric characteristics of these structures. We used numerical experiments to quantify the sensitivity of our detection method, and we demonstrated its usefulness by deconvolving images of the hearing organ acquired in vitro and in vivo.     

Comparison of I5M and 4Pi-microscopy

The axial (z‐) resolution of ～100 nm provided by 4Pi and I⁵M fluorescence microscopy relies on the coherent addition of spherical wavefronts of two opposing high aperture angle lenses. Both microscopes feature a point‐spread function (PSF) with a sharp central spot that is accompanied by axially shifted sidelobes which leads to replication artefacts in the raw image data. In a 4Pi‐microscope the sidelobes are less pronounced than in I⁵M and without relevant lateral (x,y) substructure, making their posterior removal in the image reliable and fast. On the other hand, high speeds of raw data acquisition are more easily gained by I⁵M. Moreover, I⁵M features a stronger signal as compared to the commonly employed two‐photon excitation (2PE) 4Pi‐imaging mode. We investigate here the capability of both techniques to image (aqueous) specimens without artefacts. To this end, we consider the optical transfer function (OTF) of the two microscopes in conjunction with the signal‐to‐noise‐ratio (SNR) of the object to be imaged. The imaging of E. coli bacteria with an interconvertable setup enabled a direct comparison of the two imaging modes. As both systems rely on high aperture angles, water‐immersion lenses of the largest numerical aperture available (NA = 1.2) were employed. The experimental results are corroborated by simulations assuming the signal strength encountered in the experiment. The comparison of the theoretical with the experimental PSFs/OTFs showed that our setup operated close to theory in both imaging modes. Although I⁵M provided about 10 times brighter raw image data as compared to (2PE) 4Pi‐microscopy, the I⁵M data could not be entirely cleared of artefacts. In conclusion, with the current aperture angles and fluorescence signal strengths, it is not advisable to trade in the suppression of the sidelobes for a larger image signal.     

3‐D PSF fitting for fluorescence microscopy: implementation and localization application

Localization microscopy relies on computationally efficient Gaussian approximations of the point spread function for the calculation of fluorophore positions. Theoretical predictions show that under specific experimental conditions, localization accuracy is significantly improved when the localization is performed using a more realistic model. Here, we show how this can be achieved by considering three‐dimensional (3‐D) point spread function models for the wide field microscope. We introduce a least‐squares point spread function fitting framework that utilizes the Gibson and Lanni model and propose a computationally efficient way for evaluating its derivative functions. We demonstrate the usefulness of the proposed approach with algorithms for particle localization and defocus estimation, both implemented as plugins for ImageJ.     

Measurement and evaluation of display scattering

Abstract— A novel approach is introduced for the simple and effective measurement and evaluation of light scattering in general, and its application to electronic displays is described. The method is based on the measurement of the lateral distribution of reflected luminance with an imaging detector. After numerical transformation of the measured luminance distribution, we obtain the characteristics of the BRDF of the sample with high directional resolution within a limited range of inclination angles centered about the specular direction. The method is easy to implement and to carry out. Due to the absence of moving parts, it exhibits only little sensitivity with respect to variations of the geometry parameters and is thus very robust. This paper summarizes the basics of the approach, establishes the relation to conventional BRDF‐measuring methods, it introduces an implementation and presents some typical results for LCD‐monitors.     

聚砜微球的制备及应用

通过W/O/W型双重乳液法制备聚砜(PSF)微球。研究聚乙烯吡咯烷酮(PVP)、油酸(OA)、吐温80,这些油相表面活性剂对微球形成的影响。并根据微球形貌的变化来检测水中的有机污染物。结果表明:利用油酸(OA)作为表面活性剂,可以得到蜂窝状聚砜微球。并且其对水中的N-甲基吡咯烷酮(NMP)有极端的敏感反应(NMP/水,1/107)。     

基于各向异性总变分的运动模糊盲复原方法

针对目前运动模糊图像盲复原算法对图像边缘中 拐角结构复原不佳这一问题,提出了一种以各向异性总变分为图像和模糊核正则项的遥感图 像盲 复原方法。为便于进行数值计算,采用交叉算法和分裂布雷格曼迭代导出了本文提出的盲复 原方法的迭 代公式。实验结果表明,与基于同向异性总变分的盲复原方法以及基于小波框架的盲复原方 法相 比,本文方法不仅能估算出较精确的点扩散函数(PSF,point sp read function),有效地 去除图像的模糊效应,而且对图像边缘结构特别是拐角边缘结构的增强有着独特的优势。     

Development of a novel starch-derived porous silica monolith for enhancing the dissolution rate of poorly water soluble drug

A novel starch-derived porous silica monolith (PSM) and porous starch foam (PSF) were developed as a carrier in order to improve the dissolution of lovastatin. PSM was prepared by the starch gel template method and PSF was prepared by the solvent exchange method. The morphology and structure of PSM and PSF were characterized by scanning electron microscopy (SEM) and nitrogen adsorption/desorption analysis. Lovastatin was loaded into PSM and PSF by immersion/solvent evaporation. Nano-pore spatial confinement effect on the drug dissolution was systematically studied by SEM, Fourier transform infrared spectroscopy (FTIR), thermogravametric analysis (TGA), x-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and in-vitro drug dissolution studies. Lovastatin adsorbed in PSM was amorphous and lovastatin absorbed on PSF was partially present as microcrystal in the pores of PSF and partially in crystalline form distributed on the surface of PSF. PSM and PSF allowed immediate release of lovastatin and enhanced the dissolution rate. These results provide important information about the mechanism of drug adsorption and release. Accordingly, PSM and PSF have a promising future as a vehicle for the oral delivery of poorly water soluble drugs. Moreover, the effect of PSM is better than that of PSF.     

聚砜超滤膜表面辐照改性研究

以聚砜为膜材料,聚乙二醇(PEG)为添加剂,采用浸入沉淀相转化法制备聚砜平板超滤膜,然后对实验制备的聚砜膜进行紫外辐照改性,考察了辐照对聚砜膜结构和性能的影响,研究了膜辐照前后断裂强度的变化,通过表面接触角测定仪、红外分析仪(FTIR)、扫描电镜(SEM)、改性膜的纯水通量和截留性能等方法来表征膜辐照前后的结构和性能变化.研究表明:聚砜膜断裂强度经辐照后变小,但降低幅度不大;在乙醇溶液中紫外辐照后,开始纯水通量提高而截留率降低,但降低幅度不大,时间增加到一定时通量又降低,截留率变化也不大;表面接触角随辐照时间的增长而减小,表明改性后膜表面的亲水性得到明显改善;通过FTIR分析证实在聚砜膜表面产生了新的官能团;SEM实验证实反应只发生在聚砜膜的表面而非膜内部.聚砜超滤膜经紫外辐照改性取得了良好的效果.     

PAA—PSF交联复合膜对有机物水溶液反渗透分离性能的研究

本文研究了ＰＡＡ－ＰＳＦ交联复合膜对有机醇，酸，胺，醛，酮，脂等水溶液的反渗透分离性能。结果表明，ＰＡＡ－ＰＳＦ交承复合膜对有机醇，胺和醛类水溶液有很很好的反渗透分离性能，对乙醇，乙胺和乙醛水溶液的截留率分别为６６．２％，５１％和８４％，且随分子量的增大，截留率不断上升。     

基于LMS自适应算法的图像去模糊研究

传统单幅图像去模糊方法需要稀疏先验约束,导致计算量较大。为此,在自适应最小均方误差(LMS)算法的基础上,提出一种点扩散函数(PSF)估计方法。利用模糊图像得到有效突出边缘,作为自适应滤波器的输入信号,并将模糊图像作为滤波器的期望信号,用以估计PSF。在非盲去卷积过程中,采用各项异性正规化方法对清晰图像进行约束,以减少恢复图像的振铃效应。实验结果表明,该方法不需要先验约束,对运动和非运动模糊图像均可适用,在保留图像细节的同时能抑制平滑区域的噪声。