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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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Hauptverfasser: Petrich, Annett (VerfasserIn) , Hwang, Gyu Min (VerfasserIn) , La Rocca, Laetitia (VerfasserIn) , Hassan, Mariam (VerfasserIn) , Anders-Ößwein, Maria (VerfasserIn) , Sonntag-Buck, Vera (VerfasserIn) , Heuser, Anke-Mareil (VerfasserIn) , Laketa, Vibor (VerfasserIn) , Müller, Barbara (VerfasserIn) , Kräusslich, Hans-Georg (VerfasserIn) , Klaus, Severina (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 15 October 2024
In: Viruses
Year: 2024, Jahrgang: 16, Heft: 10, Pages: 1-26
ISSN:1999-4915
DOI:10.3390/v16101610
Online-Zugang:Verlag, kostenfrei, Volltext: https://doi.org/10.3390/v16101610
Verlag, kostenfrei, Volltext: https://www.mdpi.com/1999-4915/16/10/1610
Volltext
Verfasserangaben: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
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Zusammenfassung: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.
Beschreibung:Gesehen am 24.03.2025
Beschreibung:Online Resource
ISSN:1999-4915
DOI:10.3390/v16101610

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