Improvements to white light optical imaging of widely spaced, high aspect ratio nanostructures are demonstrated using dark-field field microscopy. 1D models of bright- and dark-field imaging are developed from rigorous modal diffraction theory by assuming that features are periodic. A simple model is developed to explain dark field results and simulated line images obtained using the two modalities are compared for different dimensions and materials. Increased contrast between etched features and the substrate is demonstrated in dark field, due to its reduced sensitivity to scattering from flat areas. The results are verified using silicon nanostructures fabricated by sidewall transfer lithography, and feature separation with improved tolerance to apparent substrate brightness is demonstrated during image segmentation using the Otsu method.
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http://dx.doi.org/10.1088/1361-6528/ab13de | DOI Listing |
Oblique plane microcopy (OPM), a variant of light-sheet fluorescence microscopy (LSFM), enables rapid volumetric imaging without mechanically scanning the sample or an objective. In an OPM, the sample space is mapped to a distortion free image space via remote focusing, and the oblique light-sheet plane is mapped via a tilted tertiary imaging system onto a camera. As a result, the 3D point-spread function and optical transfer function are tilted to the optical axis of the tertiary imaging system.
View Article and Find Full Text PDFFluorescent reporters for glutamate release and postsynaptic Ca signaling are essential tools for quantifying synapse functional heterogeneity across neurons and circuits. However, leveraging these probes for neuroscience requires scalable experimental frameworks. Here, we devised a high-throughput approach to efficiently collect and analyze hundreds of optical recordings of glutaamate release activity at presynaptic boutons in cultured rat hippocampal neurons.
View Article and Find Full Text PDFUnlabelled: Patient-derived cancer organoids (PDCOs) are a valuable model to recapitulate human disease in culture with important implications for drug development. However, current methods for assessing PDCOs are limited. Label-free imaging methods are a promising tool to measure organoid level heterogeneity and rapidly screen drug response in PDCOs.
View Article and Find Full Text PDFGenetically encoded calcium (Ca ) indicators (GECIs) are widely used for imaging neuronal activity, yet current limitations of existing red fluorescent GECIs have constrained their applicability. The inherently dim fluorescence and low signal-to-noise ratio of red-shifted GECIs have posed significant challenges. More critically, several red-fluorescent GECIs exhibit photoswitching when exposed to blue light, thereby limiting their applicability in all- optical experimental approaches.
View Article and Find Full Text PDFDeveloping populations of connected neurons often share spatial and/or temporal features that anticipate their assembly. A unifying spatiotemporal motif might link sensory, central, and motor populations that comprise an entire circuit. In the sensorimotor reflex circuit that stabilizes vertebrate gaze, central and motor partners are paired in time (birthdate) and space (dorso-ventral).
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