Super-resolution two-photon microscopy via scanning patterned illumination

Urban, Ben E.; Yi, Ji; Chen, Siyu; Dong, Biqin; Zhu, Yongling; DeVries, Steven H.; Backman, Vadim; Zhang, Hao F.

doi:10.1103/physreve.91.042703

Cited by 34 publications

(26 citation statements)

References 39 publications

(50 reference statements)

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“…As shown in Figures 4 and 5, these resolution gains help to not only see smaller objects, but also to resolve interactions between weakly associating components in dense environments. Super-resolution modalities are being combined to yield novel methodologies, such as structured illumination microscopy with structured photoactivation 236 , light sheet microscopy with two photon excitation 237 , and structured illumination microscopy with two photon excitation 238 , allowing for improved spatial resolution and the ability to image deep into thick samples. Finally, there have been large improvements in the technology for localizing fluorophores in the third dimension.…”

Section: Future Outlookmentioning

confidence: 99%

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

2017

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Lipids and the membranes they form are fundamental building blocks of cellular life, and their geometry and chemical properties distinguish membranes from other cellular environments. Collective processes occurring within membranes strongly impact cellular behavior and biochemistry, and understanding these processes presents unique challenges due to the often complex and myriad interactions between membrane components. Super-resolution microscopy offers a significant gain in resolution over traditional optical microscopy, enabling the localization of individual molecules even in densely labeled samples and in cellular and tissue environments. These microscopy techniques have been used to examine the organization and dynamics of plasma membrane components, providing insight into the fundamental interactions that determine membrane functions. Here, we broadly introduce the structure and organization of the mammalian plasma membrane and review recent applications of super-resolution microscopy to the study of membranes. We then highlight some inherent challenges faced when using super-resolution microscopy to study membranes, and we discuss recent technical advancements that promise further improvements to super-resolution microscopy and its application to the plasma membrane.

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Section: Future Outlookmentioning

confidence: 99%

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

2017

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“…Super-resolution optical imaging techniques, such as stochastic optical reconstruction microscopy (STORM)12, photoactivated localization microscopy (PALM)13, stimulated emission depletion (STED)141516, and structured illumination microscopy (SIM)171819, extend the ability to study sub-diffraction-limited features that were previously thought to be unresolvable by optical microscopy20. These technologies can reach spatial resolutions higher than 10 nm.…”

mentioning

confidence: 99%

Subsurface Super-resolution Imaging of Unstained Polymer Nanostructures

Urban

Dong

Nguyen

et al. 2016

Sci Rep

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Optical imaging has offered unique advantages in material researches, such as spectroscopy and lifetime measurements of deeply embedded materials, which cannot be matched using electron or scanning-probe microscopy. Unfortunately, conventional optical imaging cannot provide the spatial resolutions necessary for many nanoscopic studies. Despite recent rapid progress, super-resolution optical imaging has yet to be widely applied to non-biological materials. Herein we describe a method for nanoscopic optical imaging of buried polymer nanostructures without the need for extrinsic staining. We observed intrinsic stochastic fluorescence emission or blinking from unstained polymers and performed spatial-temporal spectral analysis to investigate its origin. We further applied photon localization super-resolution imaging reconstruction to the detected stochastic blinking, and achieved a spatial resolution of at least 100 nm, which corresponds to a six-fold increase over the optical diffraction limit. This work demonstrates the potential for studying the static heterogeneities of intrinsic polymer molecular-specific properties at sub-diffraction-limited optical resolutions.

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“…The EMCCD exposure duration matched one patterned illumination. We acquired images corresponding to 9 illumination patterns with 3 angles and 3 phases for reconstruction.…”

Section: Methodsmentioning

confidence: 99%

“…We previously reported an imaging technique, two‐photon scanning patterned illumination microscopy, which employs patterned 2P laser excitation and frequency domain SIM reconstruction to increase optical resolution and imaging depth of SRLM . Although this method simultaneously addressed some of the aforementioned drawbacks, it was limited in dynamic imaging applications due to its modulation implementation, which constrained resolution, contrast and imaging rate (see Appendix S1, Supporting Information).…”

Section: Introductionmentioning

confidence: 99%

Imaging neuronal structure dynamics using 2‐photon super‐resolution patterned excitation reconstruction microscopy

Urban

Xiao

Dong

et al. 2017

Journal of Biophotonics

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Visualizing fine neuronal structures deep inside strongly light-scattering brain tissue remains a challenge in neuroscience. Recent nanoscopy techniques have reached the necessary resolution but often suffer from limited imaging depth, long imaging time or high light fluence requirements. Here, we present two-photon super-resolution patterned excitation reconstruction (2P-SuPER) microscopy for 3-dimensional imaging of dendritic spine dynamics at a maximum demonstrated imaging depth of 130 μm in living brain tissue with approximately 100 nm spatial resolution. We confirmed 2P-SuPER resolution using fluorescence nanoparticle and quantum dot phantoms and imaged spiny neurons in acute brain slices. We induced hippocampal plasticity and showed that 2P-SuPER can resolve increases in dendritic spine head sizes on CA1 pyramidal neurons following theta-burst stimulation of Schaffer collateral axons. 2P-SuPER further revealed nanoscopic increases in dendritic spine neck widths, a feature of synaptic plasticity that has not been thoroughly investigated due to the combined limit of resolution and penetration depth in existing imaging technologies.

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Super-resolution two-photon microscopy via scanning patterned illumination

Cited by 34 publications

References 39 publications

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

Subsurface Super-resolution Imaging of Unstained Polymer Nanostructures

Imaging neuronal structure dynamics using 2‐photon super‐resolution patterned excitation reconstruction microscopy

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