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Ultima two-photon laser scanning microscope

Ultima双光子激光扫描显微镜

Company: Bruker
Catalog#: Ultima 2-Photon Microscope
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Two-photon Photoactivation to Measure Histone Exchange Dynamics in Plant Root Cells
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Date:
2015-10-20
[Abstract]  Chromatin-binding proteins play a crucial role in chromatin structure and gene expression. Direct binding of chromatin proteins both maintains and regulates transcriptional states. It is therefore important to study the binding properties of these proteins in vivo within the natural environment of the nucleus. Photobleaching, photoactivation and photoconversion (photoswitching) can provide a non-invasive experimental approach to study dynamic properties of living cells and organisms. We used photoactivation to determine exchange dynamics of histone H2B in plant stem cells of the root (Rosa et al., 2014). The stem cells of the root are located in the middle of the tissue, which made it impossible to carry out photoactivation of sufficiently small and well-defined ... [摘要]  染色质结合蛋白在染色质结构和基因表达中起着至关重要的作用。染色质蛋白的直接结合维持和调节转录状态。因此,重要的是在细胞核的自然环境内研究这些蛋白质在体内的结合特性。光漂白,光活化和光转化(光电转换)可以提供非侵入性实验方法来研究活细胞和生物体的动态性质。我们使用光活化来确定根的植物干细胞中组蛋白H2B的交换动力学(Rosa等人,2014)。根的干细胞位于组织的中间,这使得不可能在共聚焦显微镜中用常规激光照射进行足够小和明确的亚细胞区域的光活化,主要是因为散射和折射效应在根组织分散焦斑并导致太大的区域的光活化。因此,我们使用双光子激活,其具有更好的固有分辨率的照明区域。这是因为激活依赖于两个或更多个光子的同时吸收,这又依赖于强度的平方(或更高的功率) - 更尖锐的峰。在本协议中,我们将描述进行双光子光活化实验和相应的图像分析的实验程序。该协议可以用于标记有在根组织中表达的可光活化GFP(PA-GFP)的核蛋白。

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