Light-sheet fluorescence expansion microscopy: fast mapping of neural circuits at super resolution

Bürgers, Jana; Rodriguez-Gatica, Juan Eduardo; Henneberger, Christian; Oeller, Marc; Ruland, Jan A.; Siebrasse, Jan Peter; Kubitscheck, Ulrich; Schwarz, Martin K.

doi:10.1117/1.nph.6.1.015005

Cited by 35 publications

(27 citation statements)

References 38 publications

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“…Expansion Microscopy (ExM; Chen et al, 2015;Chozinski et al, 2016) of EGFP-expressing astrocytes was performed as previously described (Deshpande et al, 2017;Bürgers et al, 2019;Herde et al, 2020). Briefly, 4-10-week-old animals were deeply anesthetized and transcardially perfused with 4% paraformaldehyde (PFA) in phosphate-buffered saline (PBS, pH 7.4).…”

Section: Expansion Microscopy (Exm)mentioning

confidence: 99%

Heterogeneity and Development of Fine Astrocyte Morphology Captured by Diffraction-Limited Microscopy

Minge

Domingos

Unichenko

et al. 2021

Front. Cell. Neurosci.

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The fine processes of single astrocytes can contact many thousands of synapses whose function they can modulate through bi-directional signaling. The spatial arrangement of astrocytic processes and neuronal structures is relevant for such interactions and for the support of neuronal signaling by astrocytes. At the same time, the geometry of perisynaptic astrocyte processes is variable and dynamically regulated. Studying these fine astrocyte processes represents a technical challenge, because many of them cannot be fully resolved by diffraction-limited microscopy. Therefore, we have established two indirect parameters of astrocyte morphology, which, while not fully resolving local geometry by design, provide statistical measures of astrocyte morphology: the fraction of tissue volume that astrocytes occupy and the density of resolvable astrocytic processes. Both are straightforward to obtain using widely available microscopy techniques. We here present the approach and demonstrate its robustness across various experimental conditions using mainly two-photon excitation fluorescence microscopy in acute slices and in vivo as well as modeling. Using these indirect measures allowed us to analyze the morphology of relatively large populations of astrocytes. Doing so we captured the heterogeneity of astrocytes within and between the layers of the hippocampal CA1 region and the developmental profile of astrocyte morphology. This demonstrates that volume fraction (VF) and segment density are useful parameters for describing the structure of astrocytes. They are also suitable for online monitoring of astrocyte morphology with widely available microscopy techniques.

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Section: Expansion Microscopy (Exm)mentioning

confidence: 99%

Heterogeneity and Development of Fine Astrocyte Morphology Captured by Diffraction-Limited Microscopy

Minge

Domingos

Unichenko

et al. 2021

Front. Cell. Neurosci.

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“…This approach was, for instance, successfully used for assessing axonal projections of small molecule-induced cortical neurons [ 154 ] and RV-based trans-synaptic tracing of host–graft innervation [ 146 ]. Notably, light sheet fluorescence imaging of cleared tissue can also be combined with novel tissue expansion protocols allowing the depiction of neuronal circuits and synaptic connections in super-resolution [ 155 ].…”

Section: Available Methods For Studying Neuronal Function and Circuitmentioning

confidence: 99%

Human stem cell-based models for studying autism spectrum disorder-related neuronal dysfunction

et al. 2020

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The controlled differentiation of pluripotent stem cells (PSCs) into neurons and glia offers a unique opportunity to study early stages of human central nervous system development under controlled conditions in vitro. With the advent of cell reprogramming and the possibility to generate induced pluripotent stem cells (iPSCs) from any individual in a scalable manner, these studies can be extended to a disease- and patient-specific level. Autism spectrum disorder (ASD) is considered a neurodevelopmental disorder, with substantial evidence pointing to early alterations in neurogenesis and network formation as key pathogenic drivers. For that reason, ASD represents an ideal candidate for stem cell-based disease modeling. Here, we provide a concise review on recent advances in the field of human iPSC-based modeling of syndromic and non-syndromic forms of ASD, with a particular focus on studies addressing neuronal dysfunction and altered connectivity. We further discuss recent efforts to translate stem cell-based disease modeling to 3D via brain organoid and cell transplantation approaches, which enable the investigation of disease mechanisms in a tissue-like context. Finally, we describe advanced tools facilitating the assessment of altered neuronal function, comment on the relevance of iPSC-based models for the assessment of pharmaceutical therapies and outline potential future routes in stem cell-based ASD research.

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“…This level of resolution was achieved by another group using an improved gel recipe (Truckenbrodt et al 2018), who notably also applied the technique to brain slices. More recently, expansion microscopy has been used in combination with light-sheet microscopy to image large regions of mouse brain or an entire Drosophila brain (Bürgers et al 2019;Gao et al 2019), indicating the potential of the technique to link synaptic architecture and neural circuits. Light-sheet microscopy increases the rate of acquisition and highlights that combining super-resolution with other imaging techniques can have real benefit.…”

Section: Expansion Microscopymentioning

confidence: 99%

Current molecular approaches to investigate pre‐synaptic dysfunction

Harper

Smillie

2021

Journal of Neurochemistry

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This is a Review for the special issue "Pre synaptic Dysfuntion and Disease". Abbreviations: abFP, acid brightening fluorescent protein; ADBE, activity-dependent bulk endocytosis; APEX, ascorbate peroxidase; BONCAT, biorthogonal noncanonical amino acid tagging; CLARITY, cleared lipid-extracted acryl-hybridized rigid immunostaining/in situ hybridization-compatible tissue hydrogel; CME, clathrin-mediated endocytosis; DAB, diaminobenzidine; dSTORM, direct stochastic optical reconstruction microscopy; EM, electron microscopy; FKBP, FK504-binding protein; FLIM, fluorescence lifetime imaging; FRB, FKBP-rapamycin binding; FRET, forster resonance energy transfer; GFP, green fluorescent protein; GRAB, G-protein coupled receptor activation based; HRP, horse radish peroxidase; iGluSnFR, intensity-based glutamate fluorescent reporter; iSnFR, intensity-based fluorescent reporter; miniSOG, mini singlet oxygen generator; mTOR, mammalian target of rapamycin; PALM, photo-activated localization microscopy; pH, potential of hydrogen; sdTIM, subdiffractional tracking of internalized molecules; SILAC, stable isotope labelling by amino acids in cell culture; SILAM, stable istope labelling in mammals; smFRET, single-molecule FRET; SNARE, soluble N-ethylmaleimide-sensitive factor Attachment Receptor; STED, stimulated emission depletion microscopy; uPAINT, universal point accumulation imaging in nanoscale topography; vGAT, vesicular gamma aminobutyric acid transporter; vGLUT, vesicular glutamate transporter.

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Light-sheet fluorescence expansion microscopy: fast mapping of neural circuits at super resolution

Cited by 35 publications

References 38 publications

Heterogeneity and Development of Fine Astrocyte Morphology Captured by Diffraction-Limited Microscopy

Heterogeneity and Development of Fine Astrocyte Morphology Captured by Diffraction-Limited Microscopy

Human stem cell-based models for studying autism spectrum disorder-related neuronal dysfunction

Current molecular approaches to investigate pre‐synaptic dysfunction

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