| Autor: |
Ying-Ying Wang, Sitong Liu, Wanzhong He, Zhaodi Jiang, Yaqi Tang, Xiangbing Qi, Gaihong Cai, Xiao-Man Liu, Xiumei Jin, Xinbin Cai, Yan Liu, Li-Lin Du, Yuhua Li, Yanyong Hu, She Chen, Ming Li, Xiaobin Sun, Ying Liu, Pei Zhao |
| Rok vydání: |
2020 |
| Předmět: |
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| DOI: |
10.21203/rs.3.pex-895/v1 |
| Popis: |
We developed a novel auto-nucleation suppressed mechanism (ANSM) for direct synthesis of EM-visible gold nanoparticles (AuNPs) on cysteine-rich tags (e.g., metallothionein) in cells for single-molecule detection with electron microscopy (it accompanies our Nature Method manuscript, Jiang et al. 2020 [1]). Both tagged-fusion proteins expressed in cells (e.g.bacteria, yeast and mammalian cells) and antigens stained with antibody-tag fusion proteins can be visualized by this protocol. Here we describe the typical protocols (both the chemical fixation and the high pressure freezing cases) developed for ANSM-based AuNP synthesis in HeLa cells expressing metallothionein (MTn) tags (Figure 1). This approach should be widely applicable to many systems for EM visualization of single-molecule in mammalian cells. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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