High-Resolution 3D Light Microscopy with STED and RESOLFT

Sahl, Steffen J.; Hell, Stefan W.

doi:10.1007/978-3-030-16638-0_1

Cited by 25 publications

(24 citation statements)

References 73 publications

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“…The fusion of viral proteins with small (19.4 kDa) and versatile tags such as SNAP or CLIP [ 99 , 100 ], that can bind these fluorophores will make it possible to optimize the labeling for each HR technique. STED microscopy provides high-speed 3D time lapse images with a lateral resolution of less than 30 nm [ 101 , 102 ]. Likewise, single molecule localization (SML) based techniques such as PALM [ 70 ] and STORM [ 64 ] have become more suited for many applications due to the implementation of high-speed CMOS cameras and “user-friendly” reconstruction and analysis software [ 103 , 104 , 105 , 106 ].…”

Section: Discussionmentioning

confidence: 99%

Imaging Viral Infection by Fluorescence Microscopy: Focus on HIV-1 Early Stage

Mukherjee

Boutant²,

Réal³

et al. 2021

Viruses

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During the last two decades, progresses in bioimaging and the development of various strategies to fluorescently label the viral components opened a wide range of possibilities to visualize the early phase of Human Immunodeficiency Virus 1 (HIV-1) life cycle directly in infected cells. After fusion of the viral envelope with the cell membrane, the viral core is released into the cytoplasm and the viral RNA (vRNA) is retro-transcribed into DNA by the reverse transcriptase. During this process, the RNA-based viral complex transforms into a pre-integration complex (PIC), composed of the viral genomic DNA (vDNA) coated with viral and host cellular proteins. The protective capsid shell disassembles during a process called uncoating. The viral genome is transported into the cell nucleus and integrates into the host cell chromatin. Unlike biochemical approaches that provide global data about the whole population of viral particles, imaging techniques enable following individual viruses on a single particle level. In this context, quantitative microscopy has brought original data shedding light on the dynamics of the viral entry into the host cell, the cytoplasmic transport, the nuclear import, and the selection of the integration site. In parallel, multi-color imaging studies have elucidated the mechanism of action of host cell factors implicated in HIV-1 viral cycle progression. In this review, we describe the labeling strategies used for HIV-1 fluorescence imaging and report on the main advancements that imaging studies have brought in the understanding of the infection mechanisms from the viral entry into the host cell until the provirus integration step.

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Section: Discussionmentioning

confidence: 99%

Imaging Viral Infection by Fluorescence Microscopy: Focus on HIV-1 Early Stage

Mukherjee

Boutant²,

Réal³

et al. 2021

Viruses

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“…The RESOLFT family [8,9,13,14] includes saturated structured illumination microscopy (S-SIM) [15], which is an upgrade of the previously structured illumination microscopy (SIM) [16]. In the classical SIM, Gustafsson implemented high-frequency line-patterned illumination and improved axial and lateral resolution by a factor of two [16,17].…”

Section: Deterministic Srm Approaches: Sim and Stedmentioning

confidence: 99%

How Single-Molecule Localization Microscopy Expanded Our Mechanistic Understanding of RNA Polymerase II Transcription

Hoboth

Šebesta

Hozák

2021

IJMS

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Classical models of gene expression were built using genetics and biochemistry. Although these approaches are powerful, they have very limited consideration of the spatial and temporal organization of gene expression. Although the spatial organization and dynamics of RNA polymerase II (RNAPII) transcription machinery have fundamental functional consequences for gene expression, its detailed studies have been abrogated by the limits of classical light microscopy for a long time. The advent of super-resolution microscopy (SRM) techniques allowed for the visualization of the RNAPII transcription machinery with nanometer resolution and millisecond precision. In this review, we summarize the recent methodological advances in SRM, focus on its application for studies of the nanoscale organization in space and time of RNAPII transcription, and discuss its consequences for the mechanistic understanding of gene expression.

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“…The RESOLFT family [8,9,13,14] includes saturated structured illumination microscopy (S-SIM) [15], which is an upgrade of previous structured illumination microscopy (SIM) [16]. In the classical SIM Gustafsson implemented high-frequency line-patterned illumination and improved axial and lateral resolution by a factor of two [16,17].…”

Section: Deterministic Srm Approaches: Sim and Stedmentioning

confidence: 99%

How Single-Molecule Localization Microscopy Expanded Our Mechanistic Understanding of RNA Polymerase II Transcription

Hoboth

Šebesta

Hozák

2021

Preprint

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Classical models of gene expression were built using genetics and biochemistry. Although these approaches are powerful, they have very limited consideration of the spatial and temporal organization of gene expression. Although the spatial organization and dynamics of RNA polymerase II (RNAPII) transcription machinery has fundamental functional consequences for gene expression, its detailed studies have been for long time abrogated by the limits of classical light microscopy. The advent of super-resolution microscopy (SRM) techniques allowed for the visualization of the RNAPII transcription machinery with nanometer resolution and millisecond precision. In this review, we summarize the recent methodological advances in SRM, focus on its application for studies of the nanoscale organization in space and time of RNAPII transcription, and discuss its consequences for the mechanistic understanding of gene expression.

show abstract

High-Resolution 3D Light Microscopy with STED and RESOLFT

Cited by 25 publications

References 73 publications

Imaging Viral Infection by Fluorescence Microscopy: Focus on HIV-1 Early Stage

Imaging Viral Infection by Fluorescence Microscopy: Focus on HIV-1 Early Stage

How Single-Molecule Localization Microscopy Expanded Our Mechanistic Understanding of RNA Polymerase II Transcription

How Single-Molecule Localization Microscopy Expanded Our Mechanistic Understanding of RNA Polymerase II Transcription

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