Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

Adam C. Parslow; Andrew H.A. Clayton; Peter Lock; Andrew M. Scott

doi:10.3791/57164

JoVE Journal
Biology

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JoVE Journal Biology

Confocal Microscopy Reveals Cell Surface Receptor Aggregation Through Image Correlation Spectroscopy

共聚焦显微镜通过图像相关谱揭示细胞表面受体聚集

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06:51 min

August 2, 2018

DOI: 10.3791/57164-v

Adam C. Parslow^1,2, Andrew H.A. Clayton³, Peter Lock⁴, Andrew M. Scott^1,2,5,6,7

¹Tumour Targeting Laboratory,Olivia Newton-John Cancer Research Institute, ²School of Cancer Medicine,La Trobe University, ³Centre for Micro-Photonics, Faculty of Science, Engineering and Technology,Swinburne University of Technology, ⁴LIMS Bioimaging Facility, La Trobe Institute for Molecular Science,La Trobe University, ⁵Department of Medical Oncology, Olivia Newton-John Cancer and Wellnes Centre,Austin Health, ⁶Department of Medicine,University of Melbourne, ⁷Department of Molecular Imaging and Therapy,Austin Health

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与细胞表面上的靶受体结合的抗体可以赋予构象和聚类改变。这些动态变化对表征靶细胞内药物的发展有影响。该协议利用共焦显微和图像相关光谱学通过 ImageJ/斐济, 以量化的程度, 受体聚类在细胞表面。

该图像相关光谱协议的总体目标是为量化细胞表面发生的聚类事件提供一种可访问的方法。与细胞表面受体结合的抗体可以赋予确认和聚集改变。对这些动态过程的分析对于药物靶标的表征非常重要。

该技术的主要优点是它提供了一种可访问的方法，用于使用易于访问的成像设备量化细胞表面的聚类事件。首先，将 10， 000 个 A431 表皮样癌细胞接种到八腔成像载玻片的每个孔中。第二天，向细胞中立即添加每毫升 100 纳克的 EGF 配体，以刺激 EGF 受体聚集。

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生物学问题 138 共焦显微镜图像相关光谱学 (ICS) 抗体: 受体相互作用抗体-药物共轭 (ADC) imageJ 斐济图像处理聚类聚集