A Next-generation Tissue Microarray (ngTMA) Protocol for Biomarker Studies
A Next-generation Tissue Microarray (ngTMA) Protocol for Biomarker Studies
复制标题
DOI：
10.3791/51893
复制
发表时间：
2014-09-01
影响因子：
1.2
通讯作者：
Lugli, Alessandro
中科院分区：
文献类型：
--
作者：
Zlobec, Inti;Suter, Guido;Lugli, Alessandro
生物标志物研究依赖于组织微阵列（TMA）。tma是通过将小组织核从“供体”块反复转移到“受体”块而产生的，然后用于各种生物标志物应用。传统tma的构建是劳动密集型的、不精确的、耗时的。在这里，概述了使用下一代组织微阵列（ngTMA）的协议。ngTMA是基于TMA规划和设计，数字病理学和自动组织微阵列。该方案是用134例转移性结直肠癌患者的例子说明。组织学，统计学和后勤方面被考虑，如组织类型，特定的组织学区域，细胞类型，组织斑点的数量，样本量，统计分析，TMA拷贝的数量。每位患者的组织学切片都被扫描并上传到基于网络的数字平台上。在那里，使用0.6-2.0 mm直径的工具查看和注释（标记），多次使用各种颜色来区分组织区域。供体块和12个“受体”块被加载到仪器中。检索数字幻灯片并与供体块图像匹配。注释区域的重复数组会自动执行，从而产生ngTMA。在本例中，规划了6个ngtma，包含6个不同的组织类型/组织学区。需要两个ngtma副本。每个病人扫描三到四张载玻片；3次扫描是必要的，并在夜间进行。所有幻灯片都有注释；用不同的颜色表示不同的组织/区域，即肿瘤中心、侵袭前沿、肿瘤/间质、淋巴结转移、肝转移和正常组织。17个注释/案例；注释时间为2-3分钟/case。共产生12个ngtma，包含4556个点。排列时间为15-20小时。由于其精度、灵活性和速度，ngTMA是进一步提高临床和转化研究中使用的tma质量的有力工具。
Biomarker research relies on tissue microarrays (TMA). TMAs are produced by repeated transfer of small tissue cores from a 'donor' block into a 'recipient' block and then used for a variety of biomarker applications. The construction of conventional TMAs is labor intensive, imprecise, and time-consuming. Here, a protocol using next-generation Tissue Microarrays (ngTMA) is outlined. ngTMA is based on TMA planning and design, digital pathology, and automated tissue microarraying. The protocol is illustrated using an example of 134 metastatic colorectal cancer patients. Histological, statistical and logistical aspects are considered, such as the tissue type, specific histological regions, and cell types for inclusion in the TMA, the number of tissue spots, sample size, statistical analysis, and number of TMA copies. Histological slides for each patient are scanned and uploaded onto a web-based digital platform. There, they are viewed and annotated (marked) using a 0.6-2.0 mm diameter tool, multiple times using various colors to distinguish tissue areas. Donor blocks and 12 'recipient' blocks are loaded into the instrument. Digital slides are retrieved and matched to donor block images. Repeated arraying of annotated regions is automatically performed resulting in an ngTMA. In this example, six ngTMAs are planned containing six different tissue types/histological zones. Two copies of the ngTMAs are desired. Three to four slides for each patient are scanned; 3 scan runs are necessary and performed overnight. All slides are annotated; different colors are used to represent the different tissues/zones, namely tumor center, invasion front, tumor/stroma, lymph node metastases, liver metastases, and normal tissue. 17 annotations/case are made; time for annotation is 2-3 min/case. 12 ngTMAs are produced containing 4,556 spots. Arraying time is 15-20 hr. Due to its precision, flexibility and speed, ngTMA is a powerful tool to further improve the quality of TMAs used in clinical and translational research.