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Recycling Endosomes and Transcytosis00:58

Recycling Endosomes and Transcytosis

The recycling endosome, also known as the endosomal recycling compartment (ERC), is a part of the slow-recycling process of the endocytic pathway. Molecules internalized through receptor-mediated endocytosis are either degraded in the lysosomes or are recycled to the plasma membrane through the fast- or slow-recycling route.
The recycling endosome is not a single organelle but an extensively tubulated network of recycling pathways. It functions in storing molecules or transporting them across...

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Membrane Transport Processes Analyzed by a Highly Parallel Nanopore Chip System at Single Protein Resolution
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Published on: August 16, 2016

Nanochannel-based single molecule recycling.

John F Lesoine1, Prahnesh A Venkataraman, Peter C Maloney

  • 1Institute of Optics, University of Rochester, Rochester, New York 14627, United States.

Nano Letters
|June 6, 2012
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Summary

This study introduces a novel method for repeatedly measuring single-molecule fluorescence without surface attachment. The technique uses electroosmosis in nanochannels, reducing susceptibility to fluorescence blinking for enhanced single-molecule analysis.

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Engineering Molecular Recognition with Bio-mimetic Polymers on Single Walled Carbon Nanotubes
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Engineering Molecular Recognition with Bio-mimetic Polymers on Single Walled Carbon Nanotubes

Published on: January 10, 2017

Area of Science:

  • Biophysics
  • Analytical Chemistry
  • Nanotechnology

Background:

  • Single-molecule fluorescence measurements are crucial for understanding molecular behavior.
  • Existing methods often require surface immobilization, which can alter molecular properties.
  • Fluorescence blinking is a common challenge in single-molecule detection.

Purpose of the Study:

  • To develop a method for repeated single-molecule fluorescence measurements without surface immobilization.
  • To overcome limitations of existing fluorescence-based trapping schemes, such as susceptibility to blinking.
  • To enable precise diffusion coefficient measurements at the single-molecule level.

Main Methods:

  • Utilizing electroosmosis to repeatedly drive single molecules through a laser focus within a fused silica nanochannel.
  • Employing detected single-molecule fluorescence to reverse electrical potential and control flow direction.
  • Integrating single-pair Förster energy transfer (spFRET) for advanced analysis.

Main Results:

  • Successfully measured single-molecule fluorescence hundreds of times without surface attachment.
  • Demonstrated the ability to recycle both proteins and DNA within nanochannels.
  • Showcased the method's reduced susceptibility to fluorescence blinking compared to existing techniques.
  • Validated diffusion coefficient measurement on a single-molecule level using turnaround time variations.

Conclusions:

  • Nanochannel-based single-molecule recycling offers a promising approach for studying molecular dynamics in solution.
  • The method allows for conformational dynamics studies on the same single molecule without surface tethering.
  • This technique enhances the reliability of single-molecule fluorescence measurements by mitigating blinking artifacts.