Cost-effective high-speed, three-dimensional live-cell imaging of HIV-1 transfer at the T cell virological synapse

Sandmeyer, Alice; Wang, Lili; Hübner, Wolfgang; Müller, Marcel; Chen, Benjamin; Huser, Thomas

doi:10.1016/j.isci.2022.105468

Cited by 2 publications

(2 citation statements)

References 46 publications

(70 reference statements)

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“…This makes the method suited for the analyses of particles bigger than 100 nm. SIM finds applications in the recording of single virus particles such as HIV which are around 100–120 nm in diameter [ 137 ], influenza virus [ 138 ] or vaccinia virus [ 139 ]. In AAV research, it can be used to visualize cellular compartments but less so single particles.…”

Section: Microscopy Methodsmentioning

confidence: 99%

Fluorescence Microscopy in Adeno-Associated Virus Research

Golm,

Hübner,

Müller

2023

Viruses

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Research on adeno-associated virus (AAV) and its recombinant vectors as well as on fluorescence microscopy imaging is rapidly progressing driven by clinical applications and new technologies, respectively. The topics converge, since high and super-resolution microscopes facilitate the study of spatial and temporal aspects of cellular virus biology. Labeling methods also evolve and diversify. We review these interdisciplinary developments and provide information on the technologies used and the biological knowledge gained. The emphasis lies on the visualization of AAV proteins by chemical fluorophores, protein fusions and antibodies as well as on methods for the detection of adeno-associated viral DNA. We add a short overview of fluorescent microscope techniques and their advantages and challenges in detecting AAV.

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Section: Microscopy Methodsmentioning

confidence: 99%

Fluorescence Microscopy in Adeno-Associated Virus Research

Golm,

Hübner,

Müller

2023

Viruses

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“…[21,22] Traditional fluorescence microscopes employ continuous illumination and camera-based image acquisition, resulting in relatively slow imaging speeds (typically between 10 and 24 fps); this factor impedes the timely acquisition of high-quality images during dynamic processes. [23,24] Although confocal microscopy employs a point scanning method, it still falls short of fulfilling the real-time imaging requirements for certain fast dynamic processes. [25] Moreover, the resolution of conventional imaging equipment (above 200 nm) is limited by the Abbe diffraction limit, thereby hindering accurate observation of nanoscale structures, particularly interorganelle membrane contacts.…”

Section: Introductionmentioning

confidence: 99%

Ion Monitoring at Nanoscale Sites of Interorganelle Membrane Contact in Living Cells

Sun,

Wang,

Zhang

et al. 2023

Small Structures

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Nanostructural contact sites formed by interorganelle membrane contacts, including mitochondria and lysosome contacts (MLC), facilitate the exchange of substances during various life processes. However, existing bioanalytical technologies have yet to provide accurate information on the exchange of substances, as these techniques exhibit limited spatial and temporal resolution. To address this limitation, a strategy is proposed that combines fluorescence resonance energy transfer (FRET) probes with high spatial resolution detection and super‐resolution microscopes (SRM) that feature time‐resolved imaging. Specifically, a proof‐of‐concept approach is presented for monitoring H+ fluctuations during MLC with a spatial H+ biosensor targeting lysosomes, BDP‐RhB. The biosensor comprises H+‐sensitive rhodamine B as a FRET acceptor connected by a flexible chain to a BODIPY derivative as a donor. The acidity of MLC sites may vary, influencing the spatial distance of the flexible chain and causing a fluorescence transition in BDP‐RhB. Consequently, the spatial distribution of H+ can be identified using SRM. Furthermore, an algorithm has been developed to screen and identify potential compounds that control substance exchange in the MLC. Collectively, this work presents the dynamic of H+ in lysosomes within living cells, which provides a drug screening tool for studying substance exchange through interorganelle membrane contacts.

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Cost-effective high-speed, three-dimensional live-cell imaging of HIV-1 transfer at the T cell virological synapse

Cited by 2 publications

References 46 publications

Fluorescence Microscopy in Adeno-Associated Virus Research

Fluorescence Microscopy in Adeno-Associated Virus Research

Ion Monitoring at Nanoscale Sites of Interorganelle Membrane Contact in Living Cells

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