Publications by authors named "Richard W Allen"

Article Synopsis
  • Protein sorting is crucial in neurotransmission, affecting the makeup of synaptic vesicles that release neurotransmitters.
  • This study used a single molecule quantification technique to analyze the variability in the number of seven membrane proteins in synaptic vesicles.
  • Results showed that some proteins were consistently sorted with high precision, while others displayed significant variability, suggesting that changes in protein expression could impact vesicle function.
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The size of a synaptic vesicle (SV) is generally thought to be determined by the amount of lipid and membrane protein it contains. Once formed, it is thought to remain constant in size. Using fluorescence correlation spectroscopy and cryogenic electron microscopy, we show that glutamatergic vesicles reversibly increase their size upon filling with glutamate.

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This article describes two complementary techniques, single-particle tracking and correlation spectroscopy, for accurately sizing nanoparticles confined within picoliter volume aqueous droplets. Single-particle tracking works well with bright particles that can be continuously illuminated and imaged, and we demonstrated this approach for sizing single fluorescent beads. Fluorescence correlation spectroscopy detects small intensity bursts from particles or molecules diffusing through the confocal probe volume, which works well with dim and rapidly diffusing particles or molecules; we demonstrated FCS for sizing synaptic vesicles confined in aqueous droplets.

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