[The gut, a whistleblower, in the early stages of Parkinson's disease].

The enteric nervous system (ENS), often called the "second brain", plays a crucial role in regulating digestive functions. Dysfunctions of the ENS are associated with several diseases such as Parkinson's disease. Recent studies suggest that early digestive disorders, notably chronic constipation, may be early signs of this neurodegenerative disease.

Second harmonic generation from aluminum plasmonic nanocavities: from scanning to imaging.

Metamaterials and plasmonic structures made from aluminum (Al) have attracted significant interest due to their low cost, long-term stability, and the relative abundance of aluminum compared to the rare metals. Also, aluminum displays distinct dielectric properties allowing for the excitation of surface plasmons in the ultraviolet region with minimal non-radiative losses. Despite these clear advantages, most of the research has been focused on either gold or silver, probably due to difficulties in forming smooth thin films of aluminum.

Label-free, fast, 2-photon volume imaging of the organization of neurons and glia in the enteric nervous system.

The enteric nervous system (ENS), sometimes referred to as a "second brain" is a quasi-autonomous nervous system, made up of interconnected plexuses organized in a mesh-like network lining the gastrointestinal tract. Originally described as an actor in the regulation of digestion, bowel contraction, and intestinal secretion, the implications of the ENS in various central neuropathologies has recently been demonstrated. However, with a few exceptions, the morphology and pathologic alterations of the ENS have mostly been studied on thin sections of the intestinal wall or, alternatively, in dissected explants.

Optometry for a short-sighted microscope.

Evanescent-wave scattering is a topic in classical electrodynamics and in the study of colloidal particles near a boundary. However, how such near-surface scattering at subcellular refractive-index heterogeneities degrades the excitation confinement in biological total internal reflection fluorescence microscopy has not been well studied. An elegant theoretical work by Axelrod and Axelrod now addresses this very relevant question and reveals that-even when scattered-evanescent light preserves some of its surprising optical properties.

A role for BDNF- and NMDAR-induced lysosomal recruitment of mTORC1 in the regulation of neuronal mTORC1 activity.

Memory and long term potentiation require de novo protein synthesis. A key regulator of this process is mTORC1, a complex comprising the mTOR kinase. Growth factors activate mTORC1 via a pathway involving PI3-kinase, Akt, the TSC complex and the GTPase Rheb.

A simple, inexpensive and multi-scale 3-D fluorescent test sample for optical sectioning microscopies.

Fluorescence standards allow for quality control and for the comparison of data sets across instruments and laboratories in applications of quantitative fluorescence. For example, users of microscopy core facilities can expect a homogenous and time-invariant illumination and an uniform detection sensitivity, which are prerequisites for imaging analysis, tracking or fluorimetric pH or Ca -concentration measurements. Similarly, confirming the three-dimensional (3-D) resolution of optical sectioning microscopes calls for a regular calibration with a standardized point source.

Astrocytes respond to a neurotoxic Aβ fragment with state-dependent Ca alteration and multiphasic transmitter release.

Excessive amounts of amyloid β (Aβ) peptide have been suggested to dysregulate synaptic transmission in Alzheimer's disease (AD). As a major type of glial cell in the mammalian brain, astrocytes regulate neuronal function and undergo activity alterations upon Aβ exposure. Yet the mechanistic steps underlying astrocytic responses to Aβ peptide remain to be elucidated.

Infrared Excitation Induces Heating and Calcium Microdomain Hyperactivity in Cortical Astrocytes.

Unraveling how neural networks process and represent sensory information and how these cellular signals instruct behavioral output is a main goal in neuroscience. Two-photon activation of optogenetic actuators and calcium (Ca) imaging with genetically encoded indicators allow, respectively, the all-optical stimulation and readout of activity from genetically identified cell populations. However, these techniques locally expose the brain to high near-infrared light doses, raising the concern of light-induced adverse effects on the biology under study.

Supercritical Angle Fluorescence Microscopy and Spectroscopy.

Fluorescence detection, either involving propagating or near-field emission, is widely being used in spectroscopy, sensing, and microscopy. Total internal reflection fluorescence (TIRF) confines fluorescence excitation by an evanescent (near) field, and it is a popular contrast generator for surface-selective fluorescence assays. Its emission equivalent, supercritical angle fluorescence (SAF), is comparably less established, although it achieves a similar optical sectioning as TIRF does.

Fabrication of Dipole-Aligned Thin Films of Porphyrin J-Aggregates over Large Surfaces.

We report a new approach for large-scale alignment of micron-sized J-aggregates of a derivative of porphyrin onto planar glass substrates. We applied a unidirectional nitrogen flow to an aqueous dye drop deposited onto a glass substrate to form an about 5 nm thick film of aligned J-aggregates over macroscopic surface areas up to several millimeters. The inter-aggregate distance is ∼500 nm, and it scales with the nitrogen pressure.

Fast 3-D Imaging of Brain Organoids With a New Single-Objective Planar-Illumination Two-Photon Microscope.

Human inducible pluripotent stem cells (hiPSCs) hold a large potential for disease modeling. hiPSC-derived human astrocyte and neuronal cultures permit investigations of neural signaling pathways with subcellular resolution. Combinatorial cultures, and three-dimensional (3-D) embryonic bodies (EBs) enlarge the scope of investigations to multi-cellular phenomena.

Calibrating Evanescent-Wave Penetration Depths for Biological TIRF Microscopy.

Roughly half of a cell's proteins are located at or near the plasma membrane. In this restricted space, the cell senses its environment, signals to its neighbors, and exchanges cargo through exo- and endocytotic mechanisms. Ligands bind to receptors, ions flow across channel pores, and transmitters and metabolites are transported against concentration gradients.

Decoding the Information Contained in Fluorophore Radiation Patterns.

Dipole radiation patterns change when a fluorescent molecule comes close to the boundary between media of different refractive indices. Near-interface molecules emit mostly into the higher-index medium, predominantly around the critical angle. The radiation pattern encodes information about the emitter distance, orientation, and the refractive index of the embedding medium.

Near-Membrane Refractometry Using Supercritical Angle Fluorescence.

Total internal reflection fluorescence (TIRF) microscopy and its variants are key technologies for visualizing the dynamics of single molecules or organelles in live cells. Yet truly quantitative TIRF remains problematic. One unknown hampering the interpretation of evanescent-wave excited fluorescence intensities is the undetermined cell refractive index (RI).

Local energy on demand: Are 'spontaneous' astrocytic Ca-microdomains the regulatory unit for astrocyte-neuron metabolic cooperation?

Astrocytes are a neural cell type critically involved in maintaining brain energy homeostasis as well as signaling. Like neurons, astrocytes are a heterogeneous cell population. Cortical astrocytes show a complex morphology with a highly branched aborization and numerous fine processes ensheathing the synapses of neighboring neurons, and typically extend one process connecting to blood vessels.

Dependence of descriptors of co-localization on microscope spatiotemporal resolution and the choice of regions of interest.

The advent and increasing availability of super-resolution microscopies has prompted re-searchers to re-investigate questions of co-localization and co-clustering in the hope of providing more precise and relevant data. Here, we focus on the problem of studying inter-organelle interfaces, a topic of growing interest in cell biology. We sought to identify mitochondria-associated membrane (MAM) candidate sites from dual-colour large-field super-resolution images.

FRET-Based Nanobiosensors for Imaging Intracellular Ca²⁺ and H⁺ Microdomains.

Semiconductor nanocrystals (NCs) or quantum dots (QDs) are luminous point emitters increasingly being used to tag and track biomolecules in biological/biomedical imaging. However, their intracellular use as highlighters of single-molecule localization and nanobiosensors reporting ion microdomains changes has remained a major challenge. Here, we report the design, generation and validation of FRET-based nanobiosensors for detection of intracellular Ca(2+) and H⁺ transients.

Astrocyte VAMP3 vesicles undergo Ca2+ -independent cycling and modulate glutamate transporter trafficking.

Key Points: Mouse cortical astrocytes express VAMP3 but not VAMP2. VAMP3 vesicles undergo Ca(2+) -independent exo- and endocytotic cycling at the plasma membrane. VAMP3 vesicle traffic regulates the recycling of plasma membrane glutamate transporters.

Cell-penetrating nanobiosensors for pointillistic intracellular Ca2+-transient detection.

Small-molecule chemical calcium (Ca(2+)) indicators are invaluable tools for studying intracellular signaling pathways but have severe shortcomings for detecting local Ca(2+) entry. Nanobiosensors incorporating functionalized quantum dots (QDs) have emerged as promising alternatives but their intracellular use remains a major challenge. We designed cell-penetrating FRET-based Ca(2+) nanobiosensors for the detection of local Ca(2+) concentration transients, using commercially available CANdot565QD as a donor and CaRuby, a custom red-emitting Ca(2+) indicator, as an acceptor.

New red-fluorescent calcium indicators for optogenetics, photoactivation and multi-color imaging.

Most chemical and, with only a few exceptions, all genetically encoded fluorimetric calcium (Ca(2+)) indicators (GECIs) emit green fluorescence. Many of these probes are compatible with red-emitting cell- or organelle markers. But the bulk of available fluorescent-protein constructs and transgenic animals incorporate green or yellow fluorescent protein (GFP and YFP respectively).

Eliminating unwanted far-field excitation in objective-type TIRF. Part II. combined evanescent-wave excitation and supercritical-angle fluorescence detection improves optical sectioning.

Azimuthal beam scanning makes evanescent-wave (EW) excitation isotropic, thereby producing total internal reflection fluorescence (TIRF) images that are evenly lit. However, beam spinning does not fundamentally address the problem of propagating excitation light that is contaminating objective-type TIRF. Far-field excitation depends more on the specific objective than on cell scattering.

Eliminating unwanted far-field excitation in objective-type TIRF. Part I. identifying sources of nonevanescent excitation light.

Total internal reflection fluorescence microscopy (TIRFM) achieves subdiffraction axial sectioning by confining fluorophore excitation to a thin layer close to the cell/substrate boundary. However, it is often unknown how thin this light sheet actually is. Particularly in objective-type TIRFM, large deviations from the exponential intensity decay expected for pure evanescence have been reported.

Full-field dual-color 100-nm super-resolution imaging reveals organization and dynamics of mitochondrial and ER networks.

Most structured illumination microscopes use a physical or synthetic grating that is projected into the sample plane to generate a periodic illumination pattern. Albeit simple and cost-effective, this arrangement hampers fast or multi-color acquisition, which is a critical requirement for time-lapse imaging of cellular and sub-cellular dynamics. In this study, we designed and implemented an interferometric approach allowing large-field, fast, dual-color imaging at an isotropic 100-nm resolution based on a sub-diffraction fringe pattern generated by the interference of two colliding evanescent waves.

New tools for investigating astrocyte-to-neuron communication.

Gray matter protoplasmic astrocytes extend very thin processes and establish close contacts with synapses. It has been suggested that the release of neuroactive gliotransmitters at the tripartite synapse contributes to information processing. However, the concept of calcium (Ca(2+))-dependent gliotransmitter release from astrocytes, and the release mechanisms are being debated.