Wilson L A, Heraty L, Ashford B A, Coelho S, Frangi A F, Pozo J M, Ince P G, Highley J R
Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush Campus, Midlothian, EH25 9RG, UK.
UK Dementia Research Institute, Cardiff University, HadynEllis Building, Maindy Road, Cardiff, CF24 4HQ, UK.
J Neurosci Methods. 2021 Jan 1;347:108963. doi: 10.1016/j.jneumeth.2020.108963. Epub 2020 Sep 30.
Tissue microarrays (TMAs), where each block (and thus section) contains multiple tissue cores from multiple blocks potentially allow more efficient use of tissue, reagents and time in neuropathology.
The relationship between data from TMA cores and whole sections was investigated using 'virtual' TMA cores. This involved quantitative assessments of microglial pathology in white matter lesions and motor neuron disease, alongside qualitative TDP-43 inclusion status in motor neuron disease cases. Following this, a protocol was developed for TMA construction.
For microglial pathology we found good concordance between virtual cores and whole sections for volume density using one 1.75 mm core (equivalent to a 2 mm core after accounting for peripheral tissue loss). More sophisticated microglial cell size and measures required two cores. Qualitative results of pTDP-43 pathology showed use of one 1.75 mm core gave a 100 % sensitivity and specificity within grey matter, and 88.3 % sensitivity and 100 % specificity within white matter. A method of producing the TMAs was suitable for immunohistochemistry both manually and by autostainer, with the minimal core loss from the microscope slide.
TMAs have been used infrequently in post mortem neuropathology research. However, we believe TMAs give comparable tissue assessment results and can be constructed, sectioned and stained with relative ease.
We found TMAs could be used to assess both quantitative (microglial pathology) and qualitative pathology (TDP-43 proteinopathy) with greatly reduced quantities of tissue, time and reagents. These could be used for further work to improve data acquisition efficiency.
组织微阵列(TMA)中,每个模块(以及切片)包含来自多个模块的多个组织芯,这可能使神经病理学中组织、试剂和时间的利用更高效。
使用“虚拟”TMA芯研究TMA芯数据与全切片数据之间的关系。这涉及对白质病变和运动神经元疾病中小胶质细胞病理学的定量评估,以及运动神经元疾病病例中TDP - 43包涵体状态的定性评估。在此之后,制定了TMA构建方案。
对于小胶质细胞病理学,我们发现使用一个1.75毫米的芯(考虑外周组织损失后相当于一个2毫米的芯),虚拟芯与全切片在体积密度方面具有良好的一致性。更复杂的小胶质细胞大小和测量需要两个芯。pTDP - 43病理学的定性结果显示,使用一个1.75毫米的芯在灰质内敏感性和特异性均为100%,在白质内敏感性为88.3%,特异性为100%。一种制作TMA的方法适用于手动和自动染色仪进行免疫组织化学,显微镜载玻片上的芯损失最小。
TMA在死后神经病理学研究中使用较少。然而,我们认为TMA能给出可比的组织评估结果,并且可以相对容易地构建、切片和染色。
我们发现TMA可用于评估定量(小胶质细胞病理学)和定性病理学(TDP - 43蛋白病),同时大大减少组织、时间和试剂的用量。这些可用于进一步工作以提高数据采集效率。