Computer-Assisted Digital Image Analysis to Quantify the Mass and Purity of Isolated Human Islets Before Transplantation
Computer-Assisted Digital Image Analysis to Quantify the Mass and Purity of Isolated Human Islets Before Transplantation
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DOI：
10.1097/tp.0b013e31818f671a
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发表时间：
2008-12-15
期刊：
影响因子：
6.2
通讯作者：
Bosco, Domenico
中科院分区：
文献类型：
--
作者：
Niclauss, Nadia;Sgroi, Antonino;Bosco, Domenico
背景资料。移植前准确测定胰岛纯度和质量是质量控制的重要组成部分。标准方法是基于对这些参数的人工评估，因此具有主观性且容易出错。因此，我们开发了一种计算机化的方法，旨在更客观地评估孤立的人类岛屿的数量和纯度。按标准方法从人胰腺组织分离纯化胰岛。对于每一种制剂，两个样品被双硫腙染色。一个样品按照标准程序用显微镜手动分析，另一个样品用数码摄影进行数字手动和计算机分析。使用Metamorph和ImageJ软件进行计算机分析，以自动量化胰岛的纯度和大小。使用Ricordi算法或考虑每个胰岛的单个体积来计算等效胰岛(IEQ)数。使用校准的红玻璃微球对计算机分析进行验证。当数字手工分析与计算机分析比较时，用Ricordi算法计算的总胰岛数、IEQ数和纯度的平均值相似。个体间的比较显示出良好的相关性(r(2)>；=0.89)。经标准手工分析，胰岛总数和纯度高于数字手工分析和计算机分析，IEQ数相近。IEQ数下降10%(P
Background. Accurate determination of islet purity and mass before transplantation is an essential part of quality control. The standard method is based on manual evaluation of these parameters and thus Subjective and prone to errors. Therefore, we developed a computerized approach aimed at evaluating more objectively the number and purity of isolated human islets.Methods. Islets were isolated and purified from human pancreata according to a standard method. For each preparation, two samples were dithizone stained. One sample was analyzed manually by microscopy, following the standard procedure, and the other was digitally photographed for both digital manual and Computerized analyses. Computerized analysis was performed using the MetaMorph and ImageJ softwares to automatically quantify purity and size of islets. Islet equivalent (IEQ) number was calculated using the Ricordi algorithm or considering the individual volume of each islet. Computerized analysis was validated using calibrated red glass microspheres.Results. When digital manual and computerized analyses were compared, mean values of total islet number, IEQ number calculated using the Ricordi algorithm, and purity were similar. Comparisons of individual values showed good correlations (r(2)>= 0.89). By standard manual analysis, total islet number and purity were higher and IEQ number similar compared with digital manual and computerized analyses. IEQ number was 10% lower (P