On-slide detection of enzymatic activities in selected single cells
Publication in refereed journal

香港中文大學研究人員
替代計量分析
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其它資訊
摘要With increasing recognition of the importance in addressing cell-to-cell heterogeneity for the understanding of complex biological systems, there is a growing need for assays capable of single cell analyses. In the current study, we describe the measurement of human topoisomerase I activity in single CD44 positive Caco2 cells specifically captured from a mixed population on glass slides, which were dual functionalized with anti-CD44-antibodies and specific DNA primers. On-slide lysis of captured CD44 positive cells, resulted in the release of human topoisomerase I, allowing the enzyme to circularize a specific linear DNA substrate added to the slides. The generated circles hybridized to the anchored DNA primers and acted as templates for a solid support rolling circle amplification reaction leading to the generation of long tandem repeat products that were detected at the single molecule level in a fluorescent microscope upon hybridization of fluorescent labelled probes. The on-slide detection system was demonstrated to be directly quantitative and specific towards CD44 positive cells. Moreover, it allowed reproducible detection of human topoisomerase I activity in single cells.
著者Keller JG, Tesauro C, Coletta A, Graversen AD, Ho YP, Kristensen P, Stougaard M, Knudsen BR
期刊名稱Nanoscale
出版年份2017
月份9
日期28
卷號9
期次36
出版社Royal Society of Chemistry
頁次13546 - 13553
國際標準期刊號2040-3364
電子國際標準期刊號2040-3372
語言英式英語
Web of Science 學科類別Chemistry, Multidisciplinary;Nanoscience & Nanotechnology;Materials Science, Multidisciplinary;Physics, Applied;Chemistry;Science & Technology - Other Topics;Materials Science;Physics

上次更新時間 2021-22-02 於 01:07