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Quantifying Cellular Internalization with a Fluorescent Click Sensor.

Laura I Selby1,2, Luigi Aurelio1, Daniel Yuen1

  • 1Monash Institute of Pharmaceutical Sciences, Monash University , Parkville , Victoria 3052 , Australia.

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|April 21, 2018
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Summary
This summary is machine-generated.

We developed a novel fluorescent sensor to accurately measure material uptake into cells. This tool differentiates surface-bound from internalized materials, aiding cell biology and drug delivery research.

Keywords:
click chemistryendocytosisflow cytometryfluorescenceinternalizationlive cell microscopy

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Area of Science:

  • Cell biology
  • Biotechnology
  • Drug delivery

Background:

  • Quantifying cellular material uptake is crucial for understanding cell biology and developing drug delivery systems.
  • Differentiating surface-bound from internalized material is a key challenge in fluorescence-based internalization assays.
  • Existing methods lack sensitivity, multiplexing capabilities, and high-throughput compatibility.

Purpose of the Study:

  • To develop a novel, small fluorescent sensor for quantifying cellular internalization.
  • To enable accurate differentiation between surface-bound and endocytosed materials.
  • To create a versatile tool for studying material uptake in cell biology and drug delivery.

Main Methods:

  • Development of a small fluorescent sensor conjugated to proteins.
  • Utilized a bio-orthogonal reaction between tetrazine and trans-cyclooctene to quench surface signals.
  • Applied simple conjugation chemistry for sensor attachment to various materials.
  • Validated the sensor using flow cytometry and fluorescence microscopy.

Main Results:

  • The sensor successfully differentiates between surface-bound and internalized materials.
  • The sensor is compatible with protein conjugation and various materials via simple chemistry.
  • The system demonstrated utility in flow cytometry and fluorescence microscopy for internalization studies.
  • The sensor's small size is comparable to standard fluorescent dyes.

Conclusions:

  • The developed fluorescent internalization sensor offers a sensitive and specific method for quantifying cellular uptake.
  • This tool overcomes limitations of traditional methods, enabling simultaneous use of multiple markers and high-throughput analysis.
  • The sensor is a valuable asset for advancing cell biology research and optimizing drug delivery carrier design.