用于再生研究的谐波纳米颗粒。
Harmonic nanoparticles for regenerative research.
作者信息
Ronzoni Flavio, Magouroux Thibaud, Vernet Remi, Extermann Jérôme, Crotty Darragh, Prina-Mello Adriele, Ciepielewski Daniel, Volkov Yuri, Bonacina Luigi, Wolf Jean-Pierre, Jaconi Marisa
机构信息
Department of Pathology and Immunology, Faculty of Medicine, University of Geneva.
Physics Department, GAP-Biophotonics, University of Geneva.
出版信息
J Vis Exp. 2014 May 1(87):51333. doi: 10.3791/51333.
In this visualized experiment, protocol details are provided for in vitro labeling of human embryonic stem cells (hESC) with second harmonic generation nanoparticles (HNPs). The latter are a new family of probes recently introduced for labeling biological samples for multi-photon imaging. HNPs are capable of doubling the frequency of excitation light by the nonlinear optical process of second harmonic generation with no restriction on the excitation wavelength. Multi-photon based methodologies for hESC differentiation into cardiac clusters (maintained as long term air-liquid cultures) are presented in detail. In particular, evidence on how to maximize the intense second harmonic (SH) emission of isolated HNPs during 3D monitoring of beating cardiac tissue in 3D is shown. The analysis of the resulting images to retrieve 3D displacement patterns is also detailed.
在这个可视化实验中，提供了使用二次谐波纳米颗粒（HNPs）对人类胚胎干细胞（hESC）进行体外标记的实验方案细节。后者是最近引入的用于标记生物样本以进行多光子成像的新型探针家族。HNPs能够通过二次谐波产生的非线性光学过程使激发光的频率加倍，且对激发波长没有限制。详细介绍了基于多光子的hESC分化为心脏簇（维持为长期气液培养）的方法。特别展示了在三维监测跳动的心脏组织过程中如何最大化分离的HNPs强烈二次谐波（SH）发射的证据。还详细介绍了对所得图像进行分析以检索三维位移模式的过程。
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