Quantifying cellular internalization with a fluorescent click sensor

Laura I Selby, Luigi Aurelio, Daniel Yuen, Bim Graham, Angus P.R. Johnston

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The ability to determine the amount of material endocytosed by a cell is important for our understanding of cell biology and in the design of effective carriers for drug delivery. To quantify internalization by fluorescence, the signal from material remaining on the cell surface must be differentiated from endocytosed material. Sensors for internalization offer advantages over traditional methods for achieving this as they exhibit improved sensitivity, allow for multiple fluorescent markers to be used simultaneously, and are amenable to high-throughput analysis. We have developed a small fluorescent internalization sensor, similar in size to a standard fluorescent dye, that can be conjugated to proteins and uses the rapid and highly specific bio-orthogonal reaction between a tetrazine and a trans-cyclooctene group to switch off the surface signal. The sensor can be attached to a variety of materials using simple chemistry and is compatible with flow cytometry and fluorescence microscopy, making it a useful tool to study the uptake of material into cells.

Original languageEnglish
Pages (from-to)1182-1189
Number of pages8
JournalACS Sensors
Volume3
Issue number6
DOIs
Publication statusPublished - 22 Jun 2018

Keywords

  • Click chemistry
  • Endocytosis
  • Flow cytometry
  • Fluorescence
  • Internalization
  • Live cell microscopy

Cite this

Selby, Laura I ; Aurelio, Luigi ; Yuen, Daniel ; Graham, Bim ; Johnston, Angus P.R. / Quantifying cellular internalization with a fluorescent click sensor. In: ACS Sensors. 2018 ; Vol. 3, No. 6. pp. 1182-1189.
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Quantifying cellular internalization with a fluorescent click sensor. / Selby, Laura I; Aurelio, Luigi; Yuen, Daniel; Graham, Bim; Johnston, Angus P.R.

In: ACS Sensors, Vol. 3, No. 6, 22.06.2018, p. 1182-1189.

Research output: Contribution to journalArticleResearchpeer-review

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