Advancements and Practical Considerations for Biophysical Research: Navigating the Challenges and Future of Super-resolution Microscopy

Chen, Huanhuan; Yan, Guangjie; Wen, Meng-Hsuan; Brooks, Kameron N.; Zhang, Yuteng; Huang, Pei-San; Chen, Tai-Yen

doi:10.1021/cbmi.4c00019

Cited by 2 publications

References 167 publications

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Human transporter de-oligomerization regulates copper uptake into cells

Chen,

Wen,

Chen

et al. 2024

Preprint

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Copper is an essential element involved in various biochemical processes, such as mitochondrial energy production and antioxidant defense, but improper regulation can lead to cellular toxicity and disease. Copper Transporter 1 (CTR1) plays a key role in copper uptake and maintaining cellular copper homeostasis. Although CTR1 endocytosis was previously thought to reduce copper uptake when levels are high, it was unclear how rapid regulation is achieved. Using single-molecule localization microscopy and single-molecule neighbor density assays, we discovered that excess copper induces monomerization of the wild-type trimeric CTR1 prior to endocytosis, a response blocked in the endocytosis-deficient CTR1 (M150L) mutant. This monomerization rapidly halts copper uptake and prevents copper overload. These findings reveal changes in protein oligomerization as a new paradigm of metal transport regulation, linking CTR1's structural changes to its endocytosis and copper homeostasis.

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Human transporter de-oligomerization regulates copper uptake into cells

Chen,

Wen,

Chen

et al. 2024

Preprint

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Expanding Insights: Harnessing Expansion Microscopy for Super-Resolution Analysis of HIV-1–Cell Interactions

Petrich,

Hwang,

La Rocca

et al. 2024

Viruses

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Expansion microscopy has recently emerged as an alternative technique for achieving high-resolution imaging of biological structures. Improvements in resolution are achieved by physically expanding samples through embedding in a swellable hydrogel before microscopy. However, expansion microscopy has been rarely used in the field of virology. Here, we evaluate and characterize the ultrastructure expansion microscopy (U-ExM) protocol, which facilitates approximately four-fold sample expansion, enabling the visualization of different post-entry stages of the HIV-1 life cycle, focusing on nuclear events. Our findings demonstrate that U-ExM provides robust sample expansion and preservation across different cell types, including cell-culture-adapted and primary CD4+ T-cells as well as monocyte-derived macrophages, which are known HIV-1 reservoirs. Notably, cellular targets such as nuclear bodies and the chromatin landscape remain well preserved after expansion, allowing for detailed investigation of HIV-1–cell interactions at high resolution. Our data indicate that morphologically distinct HIV-1 capsid assemblies can be differentiated within the nuclei of infected cells and that U-ExM enables detection of targets that are masked in commonly used immunofluorescence protocols. In conclusion, we advocate for U-ExM as a valuable new tool for studying virus–host interactions with enhanced spatial resolution.

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Advancements and Practical Considerations for Biophysical Research: Navigating the Challenges and Future of Super-resolution Microscopy

Cited by 2 publications

References 167 publications

Human transporter de-oligomerization regulates copper uptake into cells

Human transporter de-oligomerization regulates copper uptake into cells

Expanding Insights: Harnessing Expansion Microscopy for Super-Resolution Analysis of HIV-1–Cell Interactions

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