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High-resolution elemental mapping of human placental chorionic villi using synchrotron X-ray fluorescence spectroscopy

机译:使用同步加速器X射线荧光光谱法对人胎盘绒毛膜绒毛进行高分辨率元素定位

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摘要

The placenta is the organ that mediates transport of nutrients and waste materials between mother and fetus. Synchrotron X-ray fluorescence (SXRF) microanalysis is a tool for imaging the distribution and quantity of elements in biological tissue, which can be used to study metal transport across biological membranes. Our aims were to pilot placental biopsy specimen preparation techniques that could be integrated into an ongoing epidemiology birth cohort study without harming rates of sample acquisition. We studied the effects of fixative (formalin or glutaraldehyde) and storage duration (30 days or immediate processing) on metal distribution and abundance and investigated a thaw-fixation protocol for archived specimens stored at -80 A degrees C. We measured fixative elemental composition with and without a placental biopsy via inductively coupled plasma mass spectrometry (ICP-MS) to quantify fixative-induced elemental changes. Formalin-fixed specimens showed hemolysis of erythrocytes. The glutaraldehyde-paraformaldehyde solution in HEPES buffer (GTA-HEPES) had superior anatomical preservation, avoided hemolysis, and minimized elemental loss, although some cross-linking of exogenous Zn was evident. Elemental loss from tissue stored in fixative for 1 month showed variable losses (a parts per thousand 40 % with GTA-HEPES), suggesting storage duration be controlled for. Thawing of tissue held at -80 A degrees C in a GTA-HEPES solution provided high-quality visual images and elemental images.
机译:胎盘是介导母亲和胎儿之间营养物质和废物物质运输的器官。同步加速器X射线荧光(SXRF)微分析是一种用于对生物组织中元素的分布和数量进行成像的工具,可用于研究金属在生物膜上的传输。我们的目的是试行胎盘活检标本制备技术,这些技术可以整合到正在进行的流行病学出生队列研究中,而不会影响样本采集率。我们研究了固定剂(福尔马林或戊二醛)和储存时间(30天或立即加工)对金属分布和丰度的影响,并研究了在-80 A摄氏度下储存的标本的解冻固定方案。而且无需通过电感耦合等离子体质谱(ICP-MS)进行胎盘活检来定量固定剂引起的元素变化。福尔马林固定的标本显示红细胞溶血。 HEPES缓冲液(GTA-HEPES)中的戊二醛-多聚甲醛溶液具有出色的解剖学保存能力,避免了溶血作用,并将元素损失降至最低,尽管外源锌的某些交联是明显的。固定剂中储存1个月的组织中元素损失显示出可变的损失(GTA-HEPES为千分之一40%),表明可以控制储存时间。在GTA-HEPES解决方案中,在-80 A摄氏度下融化的组织可提供高质量的可视图像和基本图像。

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