Expanding insights: harnessing expansion microscopy for super-resolution analysis of HIV-1-cell interactions
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...
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| Main Authors: | , , , , , , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
15 October 2024
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| In: |
Viruses
Year: 2024, Volume: 16, Issue: 10, Pages: 1-26 |
| ISSN: | 1999-4915 |
| DOI: | 10.3390/v16101610 |
| Online Access: | Verlag, kostenfrei, Volltext: https://doi.org/10.3390/v16101610 Verlag, kostenfrei, Volltext: https://www.mdpi.com/1999-4915/16/10/1610 
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| Author Notes: | Annett Petrich, Gyu Min Hwang, Laetitia La Rocca, Mariam Hassan, Maria Anders-Össwein, Vera Sonntag-Buck, Anke-Mareil Heuser, Vibor Laketa, Barbara Müller, Hans-Georg Kräusslich and Severina Klaus |
| Summary: | 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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| Item Description: | Gesehen am 24.03.2025 |
| Physical Description: | Online Resource |
| ISSN: | 1999-4915 |
| DOI: | 10.3390/v16101610 |