神经生物学

In the emerging field of super-resolution microscopy, the branch of live-cell imaging is still in its infancy. Regardless of its importance for addressing relevant biological questions, live super-resolution imaging has to face several obstacles when compared to conventional ima ...

In a method termed photoactivated localization microscopy (PALM), super-resolution fluorescence imaging can be achieved through the localization of single molecules. This allows the resolution of specific proteins fused to the appropriate fluorescent protein label. Here, we ...

This chapter presents the foundations of Sted microscopy with a comparison to its generalization Resolft microscopy and to stochastic imaging methods (Palm, Storm, Fpalm, and alike). The first section reviews the advantages of optical microscopy, explains the diffraction limit, and ...

Diffraction sets a physical limit for the theoretically achievable resolution; however, it is possible to circumvent this barrier. That’s what microscopists have been doing in recent years and in many ways at once, starting the era of super-resolution in light microscopy. High-resoluti ...

Galileo Galilei invented the first microscope “occhiolino,” by combining a concave and a convex lens in 1600s. Robert Hooke and Anton van Leeuwenhoek later modified it to look at living things. Since then, light microscopy has gained immense popularity and has been pushing the limits of optical ...

In recent years, microscopy techniques have reached high sensitivities and excellent resolutions, far beyond the diffraction limit. However, images of biological specimens obtained with super-resolution instruments have the tendency of being dominated by spots. The quality or ...

The preparation of samples and the choice of appropriate labeling techniques have become instrumental for the development of light microscopy techniques with increasingly high resolution. Both localization microscopy and STED approaches require fluorophores with speci ...

Eighty years after its development, electron microscopy still represents the gold standard in terms of resolution. A major disadvantage is, however, the requirement for fixed specimens—especially in view of the numerous live fluorescence microscopy methods that have been develo ...

In theoretical investigations, we review several nonlinear optical approaches towards subdiffraction-limited resolution in label-free imaging via coherent anti-Stokes Raman scattering (CARS). Using a density matrix model and numerical integration, we investigate va ...

X-ray microscopy and tomography can provide the three-dimensional density distribution within cells and tissues without staining and slicing. In addition, chemical information—i.e. the elemental distribution—can be retrieved by X-ray spectro-microscopy based on contrast ...

Atomic force microscopy (AFM) is a powerful technique for analyzing the structure, properties, and interactions of living cells down to molecular resolution. Rather than using an incident beam as in optical and electron microscopies, AFM measures the tiny forces acting between a sharp tip ...

The cerebral vascular system services the constant demand for energy during neuronal activity in the brain. Attempts to delineate the logic of neurovascular coupling have been greatly aided by the advent of two-photon laser scanning microscopy to concurrently image blood flow and the ac ...

Microscopic in vivo measurements of cerebral oxygenation are of key importance for understanding normal cerebral energy metabolism and its dysregulation in a wide range of clinical conditions. Relevant cerebral pathologies include compromised blood perfusion following s ...

Microglial cells are the innate immune cells of the central nervous system. In the healthy adult brain “resting” ramified microglia continuously palpate their environment to monitor the integrity of and to react to any disturbance of brain homeostasis. During injury, inflammation, and ...

Mounting evidence from in vitro experiments supports bidirectional interactions between astrocytes and neurons, suggesting glial involvement in neuronal information processing in the brain. Elevation of the cytosolic calcium ion (Ca2+) concentration has been suggested to ...

Voltage-sensitive dyes can be topically applied to the surface of the brain in order to label membranes in the supragranular layers of the neocortex. Fluorescence signals from voltage-sensitive dyes in vivo correlate linearly with membrane potential of excitatory layer 2/3 neurons wi ...

Translating the advances seen recently in recording from neurons in vivo to dendritic recordings presents special difficulties. In vivo two-photon imaging of dendrites was achieved over a decade ago and is still the method of choice for recording from small dendritic compartments in sin ...

Mammalian cortical neurons integrate sensory information that arrives through numerous synaptic inputs located on their dendrites. Here we introduce an approach to identify sensory-evoked dendritic input sites in cortical neurons in vivo involving the use of two-photon calci ...

Optogenetic tools have gained popularity for enabling manipulation of specified populations of neurons on a precise temporal scale. These opsins, as the proteins are known, utilize retinal (Vitamin A) as a co-factor—in a form related to the same molecule used by the human retina as a light-sens ...

Chronic in vivo two-photon imaging of genetically encoded sensors has recently enabled the measurement of activity from the same individual neurons repeatedly in different imaging sessions over months, opening new possibilities to investigate the function and plasticity of neu ...