Imaging nanoscale nuclear structures with expansion microscopy.
Imaging nanoscale nuclear structures with expansion microscopy.
复制标题
DOI：
10.1242/jcs.259009
复制
发表时间：
2022-07-15
影响因子：
4
通讯作者：
中科院分区：
文献类型：
--
作者：
通常应用的超分辨率光学显微镜提供了对纳米级亚细胞过程的洞察。然而，成像深度、速度、吞吐量和成本仍然是重大挑战，限制了可以研究的三维(3D)纳米级过程的数量和能够进行此类分析的实验室数量。膨胀显微镜(EXM)解决了其中的许多局限性，但它在核过程成像方面的应用受到了核不等膨胀的担忧的限制。在这里，我们证明了原子核在分辨率等于或大于120-130 nm(预膨胀)时的各向同性膨胀的条件。以DNA损伤反应蛋白BRCA1、53BP1(也称为TP53BP1)和RAD51为样本，我们定量地描述了超过50,000个DNA损伤反应结构的3D纳米组织。我们展示了评估染色质调节事件的能力，并展示了四种元素的同时评估。因此，这项研究展示了EXM如何有助于纳米级核过程的研究。摘要：扩张显微镜提供了对染色质修饰物对DNA修复蛋白BRCA1、53BP1和RAD51时空组织的影响的定量洞察，分辨率为65-70 nm。
Commonly applied super-resolution light microscopies have provided insight into subcellular processes at the nanoscale. However, imaging depth, speed, throughput and cost remain significant challenges, limiting the numbers of three-dimensional (3D) nanoscale processes that can be investigated and the number of laboratories able to undertake such analysis. Expansion microscopy (ExM) solves many of these limitations, but its application to imaging nuclear processes has been constrained by concerns of unequal nuclear expansion. Here, we demonstrate the conditions for isotropic expansion of the nucleus at a resolution equal to or better than 120–130 nm (pre-expansion). Using the DNA damage response proteins BRCA1, 53BP1 (also known as TP53BP1) and RAD51 as exemplars, we quantitatively describe the 3D nanoscale organisation of over 50,000 DNA damage response structures. We demonstrate the ability to assess chromatin-regulated events and show the simultaneous assessment of four elements. This study thus demonstrates how ExM can contribute to the investigation of nanoscale nuclear processes. Summary: Expansion microscopy provides quantitative insight into the impact of chromatin modifiers on spatiotemporal organisation of the DNA repair proteins BRCA1, 53BP1 and RAD51 at a resolution of 65–70 nm.
登录
查看更多内容
影响因子：
48
作者：
Chen F;Wassie AT;Cote AJ;Sinha A;Alon S;Asano S;Daugharthy ER;Chang JB;Marblestone A;Church GM;Raj A;Boyden ES
通讯作者：
Boyden ES
光学成像。膨胀显微镜。
DOI：
10.1126/science.1260088
发表时间：
2015-01-30
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Chen F;Tillberg PW;Boyden ES
通讯作者：
Boyden ES
DOI：
10.1083/jcb.200510130
发表时间：
2006-04-24
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bekker-Jensen S;Lukas C;Kitagawa R;Melander F;Kastan MB;Bartek J;Lukas J
通讯作者：
Lukas J
影响因子：
4.6
作者：
Reindl J;Girst S;Walsh DW;Greubel C;Schwarz B;Siebenwirth C;Drexler GA;Friedl AA;Dollinger G
通讯作者：
Dollinger G
影响因子：
4
作者：
Chapman, J. Ross;Sossick, Alex J.;Jackson, Stephen P.
通讯作者：
Jackson, Stephen P.