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Review

Super-Resolution Microscopy in the Structural Analysis and Assembly Dynamics of HIV

by
Aiden Jurcenko
1,2,
Olesia Gololobova
2,3,* and
Kenneth W. Witwer
2,3,4,5,*
1
Department of Chemical and Biomolecular Engineering, Johns Hopkins University Whiting School of Engineering, Baltimore, MD 21218, USA
2
Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
3
EV Core Facility “EXCEL”, Institute for Basic Biomedical Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
4
The Richman Family Precision Medicine Center of Excellence in Alzheimer’s Disease, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
5
Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
*
Authors to whom correspondence should be addressed.
Appl. Nano 2025, 6(3), 13; https://doi.org/10.3390/applnano6030013
Submission received: 22 May 2025 / Revised: 15 July 2025 / Accepted: 26 July 2025 / Published: 31 July 2025
(This article belongs to the Collection Review Papers for Applied Nano Science and Technology)

Abstract

Super-resolution microscopy (SRM) has revolutionized our understanding of subcellular structures, including cell organelles and viruses. For human immunodeficiency virus (HIV), SRM has significantly advanced knowledge of viral structural biology and assembly dynamics. This review analyzes how SRM techniques (particularly PALM, STORM, STED, and SIM) have been applied over the past decade to study HIV structural components and assembly. By categorizing and comparing studies based on SRM methods, HIV components, and labeling strategies, we assess the strengths and limitations of each approach. Our analysis shows that PALM is most commonly used for live-cell imaging of HIV Gag, while STED is primarily used to study the viral envelope (Env). STORM and SIM have been applied to visualize various components, including Env, capsid, and matrix. Antibody labeling is prevalent in PALM and STORM studies, targeting Env and capsid, whereas fluorescent protein labeling is mainly associated with PALM and focused on Gag. A recent emphasis on Gag and Env points to deeper investigation into HIV assembly and viral membrane dynamics. Insights from SRM studies of HIV not only enhance virological understanding but also inform future research in therapeutic strategies and delivery systems, including extracellular vesicles.
Keywords: HIV; super-resolution microscopy; single-molecule localization microscopy; stimulated emission depletion microscopy; structured illumination microscopy; fluorescent labeling HIV; super-resolution microscopy; single-molecule localization microscopy; stimulated emission depletion microscopy; structured illumination microscopy; fluorescent labeling

Share and Cite

MDPI and ACS Style

Jurcenko, A.; Gololobova, O.; Witwer, K.W. Super-Resolution Microscopy in the Structural Analysis and Assembly Dynamics of HIV. Appl. Nano 2025, 6, 13. https://doi.org/10.3390/applnano6030013

AMA Style

Jurcenko A, Gololobova O, Witwer KW. Super-Resolution Microscopy in the Structural Analysis and Assembly Dynamics of HIV. Applied Nano. 2025; 6(3):13. https://doi.org/10.3390/applnano6030013

Chicago/Turabian Style

Jurcenko, Aiden, Olesia Gololobova, and Kenneth W. Witwer. 2025. "Super-Resolution Microscopy in the Structural Analysis and Assembly Dynamics of HIV" Applied Nano 6, no. 3: 13. https://doi.org/10.3390/applnano6030013

APA Style

Jurcenko, A., Gololobova, O., & Witwer, K. W. (2025). Super-Resolution Microscopy in the Structural Analysis and Assembly Dynamics of HIV. Applied Nano, 6(3), 13. https://doi.org/10.3390/applnano6030013

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