Publications by authors named "Daniel Flickinger"

Article Synopsis
  • Two-photon microscopy and fluorescent proteins are important tools in neuroscience, but their optimization for two-photon excitation is necessary for better experimental results.
  • Directed evolution of fluorescent proteins is typically conducted under one-photon excitation, which may not predict two-photon properties accurately.
  • The GIZMO instrument and software enable rapid screening of up to 10,000 fluorescent protein colonies for two-photon excitation in just 7 hours, enhancing the process of evolving these proteins.
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The active properties of dendrites can support local nonlinear operations, but previous imaging and electrophysiological measurements have produced conflicting views regarding the prevalence and selectivity of local nonlinearities in vivo. We imaged calcium signals in pyramidal cell dendrites in the motor cortex of mice performing a tactile decision task. A custom microscope allowed us to image the soma and up to 300 μm of contiguous dendrite at 15 Hz, while resolving individual spines.

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An amendment to this paper has been published and can be accessed via a link at the top of the paper.

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Point-scanning two-photon microscopy enables high-resolution imaging within scattering specimens such as the mammalian brain, but sequential acquisition of voxels fundamentally limits its speed. We developed a two-photon imaging technique that scans lines of excitation across a focal plane at multiple angles and computationally recovers high-resolution images, attaining voxel rates of over 1 billion Hz in structured samples. Using a static image as a prior for recording neural activity, we imaged visually evoked and spontaneous glutamate release across hundreds of dendritic spines in mice at depths over 250 µm and frame rates over 1 kHz.

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Imaging is used to map activity across populations of neurons. Microscopes with cellular resolution have small (.

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We present a bichromatic prism pair interferometer (BPPI) for controlling the delay between laser pulses of two different frequencies propagating collinearly in a single beam. The BPPI is especially useful when working with ultrafast laser pulses because it intrinsically allows for independent control over the second-order dispersion experienced by the differently colored pulses. We use this control to demonstrate successful precompensation for blue (lambda approximately 390 nm) and UV (lambda approximately 260 nm) pulses that pass through 2.

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