MINFLUX fluorescence nanoscopy in biological tissue

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: July 8, 2024

Abstract Optical imaging access to nanometer-level protein distributions in intact tissue is a highly sought-after goal, as it would provide visualization physiologically relevant contexts. Under the unfavorable signal-to-background conditions of increased absorption and scattering excitation fluorescence light complex medium, super-resolution microscopy methods are severely challenged attaining precise localization molecules. We reasoned that typical use confocal detection pinhole MINFLUX nanoscopy, suppressing background providing optical sectioning, should facilitate resolution single fluorophores even amid optically challenging environments. Here, we investigated performance for different synaptic targets fluorescent labels sections mouse brain. Single were localized with precision < 5 nm at up 80 µm sample depth. two color channels allowed probe PSD95 relative spine head morphology, while also visualizing presynaptic VGlut clustering AMPA receptor post-synapse. Our two-dimensional (2D) three-dimensional (3D) two-color results tissue, 10 3D fluorophore localization, open new avenues investigate on single-synapse level fixed living brain slices.

Language: Английский

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MINFLUX fluorescence nanoscopy in biological tissue

Proceedings of the National Academy of Sciences, Journal Year: 2024, Volume and Issue: 121(52)

Published: Dec. 20, 2024

Optical imaging access to nanometer-level protein distributions in intact tissue is a highly sought-after goal, as it would provide visualization physiologically relevant contexts. Under the unfavorable signal-to-background conditions of increased absorption and scattering excitation fluorescence light complex sample, superresolution microscopy methods are severely challenged attaining precise localization molecules. We reasoned that typical use confocal detection pinhole MINFLUX nanoscopy, suppressing background providing optical sectioning, should facilitate resolution single fluorophores even amid optically challenging environments. Here, we investigated performance for different synaptic targets fluorescent labels sections mouse brain. Single were localized with precision <5 nm at up 80 µm sample depth. two color channels allowed probe PSD95 relative spine head morphology, while also visualizing presynaptic vesicular glutamate transporter (VGlut) 1 clustering α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) postsynapse. Our two-dimensional (2D) three-dimensional (3D) two-color results tissue, <10 3D fluorophore localization, open broad avenues investigate on single-synapse level fixed living brain slices.

Language: Английский

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Squid: Simplifying Quantitative Imaging Platform Development and Deployment

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2020, Volume and Issue: unknown

Published: Dec. 29, 2020

Abstract With rapid developments in microscopy methods, highly versatile, robust and affordable implementations are needed to enable wide adoption by the biological sciences community. Here we report Squid, a quantitative imaging platform with full suite of hardware software components configurations for deploying facility-grade widefield microscopes advanced features like flat field fluorescence excitation, patterned illumination tracking microscopy, at fraction cost commercial solutions. The open modular nature (both software) lowers barrier deployment, importantly, simplifies development, making system configurable experiments that can run on easily programmable. Developed goal helping translate advances microscopy-enabled including those powered deep learning, envision Squid will simplify roll-out microscopy-based applications - point care low resource settings, make new or otherwise techniques easier, significantly increase available microscope-hours labs.

Language: Английский

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Super-resolution FRET measurements

Nanoscale, Journal Year: 2021, Volume and Issue: 13(44), P. 18421 - 18433

Published: Jan. 1, 2021

Recent advances combining FRET and super-resolution techniques are presented. This merger of methods is opening exciting opportunities to study the spatial organization functional state (bio)molecules materials at nanoscale.

Language: Английский

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Nanoparticles for super-resolution microscopy: intracellular delivery and molecular targeting

Chemical Society Reviews, Journal Year: 2022, Volume and Issue: 51(24), P. 9882 - 9916

Published: Jan. 1, 2022

Following an overview of the approaches and techniques used to acheive super-resolution microscopy, this review presents advantages supplied by nanoparticle based probes for these applications.

Language: Английский

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