Automated Digital Image Analysis of Islet Cell Mass Using Nikon's Inverted Eclipse Ti Microscope and Software to Improve Engraftment May Help to Advance the Therapeutic Efficacy and Accessibility of Islet Transplantation Across Centers
Automated Digital Image Analysis of Islet Cell Mass Using Nikon's Inverted Eclipse Ti Microscope and Software to Improve Engraftment May Help to Advance the Therapeutic Efficacy and Accessibility of Islet Transplantation Across Centers
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DOI：
10.3727/096368913x667493
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发表时间：
2015-01-01
影响因子：
3.3
通讯作者：
Kerr-Conte, Julie
中科院分区：
文献类型：
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作者：
Gmyr, Valery;Bonner, Caroline;Kerr-Conte, Julie
在将胰岛制剂移植到1型糖尿病患者体内之前，必须对胰岛活性、质量和纯度进行可靠的评估。定量人类胰岛制剂的标准方法是对双硫腙染色的胰岛样本进行直接显微镜分析，但这种技术可能会受到观察者间/观察者内变异性的影响，这可能会导致假阳性/阴性胰岛计数。在这里，我们描述了一种简单、可靠、自动化的数字图像分析(ADIA)技术，用于在胰岛移植前准确地量化胰岛总数、胰岛等效数(IEQ)和胰岛纯度。根据Ricordi等人的自动化方法，从n=42人的胰腺组织中分离纯化胰岛。每种制备方法取三个胰岛样品，用双硫腙染色，用IEQ数表示。用显微镜手动分析胰岛，或使用尼康内置MS-Elements高级研究(AR)软件的倒置日食钛显微镜自动定量胰岛。与标准手工法相比，AIDA法显著增加了符合植入条件的胰岛制剂的数量(p<；0.001)。各方法的比较表明，IEQ数的平均值(r(2)=0.91)与总的胰岛数(r(2)=0.88)有很好的相关性，用IEQ数估算胰岛表面积时，IEQ数的平均值增加到r(2)=0.93。与标准手工法相比，ADIA法具有很高的观察者内重复性(p
Reliable assessment of islet viability, mass, and purity must be met prior to transplanting an islet preparation into patients with type 1 diabetes. The standard method for quantifying human islet preparations is by direct microscopic analysis of dithizone-stained islet samples, but this technique may be susceptible to inter-/intraobserver variability, which may induce false positive/negative islet counts. Here we describe a simple, reliable, automated digital image analysis (ADIA) technique for accurately quantifying islets into total islet number, islet equivalent number (IEQ), and islet purity before islet transplantation. Islets were isolated and purified from n = 42 human pancreata according to the automated method of Ricordi et al. For each preparation, three islet samples were stained with dithizone and expressed as IEQ number. Islets were analyzed manually by microscopy or automatically quantified using Nikon's inverted Eclipse Ti microscope with built-in MS-Elements Advanced Research (AR) software. The AIDA method significantly enhanced the number of islet preparations eligible for engraftment compared to the standard manual method (p < 0.001). Comparisons of individual methods showed good correlations between mean values of IEQ number (r(2)=0.91) and total islet number (r(2)=0.88) and thus increased to r(2)=0.93 when islet surface area was estimated comparatively with IEQ number. The ADIA method showed very high intraobserver reproducibility compared to the standard manual method (p