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这是最新一期cytometry A上的一篇文章,摘要翻译如下(以下内容在60天内仅注册会员可见,60天后所有人可见):
利用完整细胞对带电荷的荧光染料拒染来监测细胞活力已经非常普遍。本文作者在研究非侵袭性并且可长期实时监测细胞活力的染料时,发现了蒽环类衍生物DRAQ7。这类染料在细胞膜完整时不会穿透细胞膜,仅当细胞膜完整性受到损伤时,才会进入细胞并与细胞核内DNA结合。
用扫描电镜和流式细胞术检测,发现DRAQ7作用72小时对很多肿瘤细胞株均无毒性,亦无诱导DNA损伤。非常适用于低氧、饥饿、药物等诱导的细胞毒性研究,并且不影响抗癌药物的作用。
而且,结合DRAQ7和线粒体膜内电位染料TMRM,作者还首次介绍了近实时微流式监测方法。
全文见二楼回帖
Cytometry A. 2012 Nov 16. doi: 10.1002/cyto.a.22228. [Epub ahead of print]
Real-time cell viability assays using a new anthracycline derivative DRAQ7®
Akagi J, Kordon M, Zhao H, Matuszek A, Dobrucki J, Errington R, J Smith P, Takeda K, Darzynkiewicz Z, Wlodkowic D.
Source
The BioMEMS Research Group, School of Chemical Sciences, University of Auckland, Auckland, New Zealand.
Abstract
The exclusion of charged fluorescent dyes by intact cells has become a well-established assay for determining viability of cells. In search for a noninvasive fluorescent probe capable of long-term monitoring of cell death in real-time, we evaluated a new anthracycline derivative DRAQ7. The novel probe does not penetrate the plasma membrane of living cells but when the membrane integrity is compromised, it enters and binds readily to nuclear DNA to report cell death. It proved to be nontoxic to a panel of cancer cell lines grown continuously for up to 72 h and did not induce any detectable DNA damage signaling when analyzed using laser scanning microscopy and flow cytometry. The DRAQ7 provided a sensitive, real-time readout of cell death induced by a variety of stressors such as hypoxia, starvation, and drug-induced cytotoxicity. The overall responses to anticancer agents and resulting pharmacological dose-response profiles were not affected by the growth of tumor cells in the presence DRAQ7. Moreover, we for the first time introduced a near real-time microflow cytometric assay based on combination of DRAQ7 and mitochondrial inner membrane potential (ΔΨ(m) ) sensitive probe TMRM. We provide evidence that this low-dosage, real-time labeling procedure provides multiparameter and kinetic fingerprint of anticancer drug action. © 2012 International Society for Advancement of Cytometry.
Copyright © 2012 International Society for Advancement of Cytometry.
PMID: 23165976 [PubMed - as supplied by publisher]
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