Delivery of Fluorescent Probes Using Streptolysin O for Fluorescence Microscopy of Living Cells

Kai Wen Teng, Pin Ren, Paul R Selvin

Research output: Contribution to journalArticle

Abstract

Methods to efficiently deliver fluorophores across the cell membrane are crucial for imaging the dynamics of intracellular proteins using fluorescence. Here we describe a simple protocol for permeabilizing living cells using streptolysin O, a bacterial toxin, which allows transient uptake of fluorescent probes for labeling specific intracellular proteins. The technique is applicable for delivering different classes of fluorescent probes with a molecular weight of <150 kDa, and it is also applicable to a variety of different cell lines. The technique enables the utilization of a broad range of fluorophores for live cell imaging of intracellular proteins. Extended observation of intracellular fluorescence bound to specific proteins is now possible through super-resolution microscopy by using fluorophores that are photostable in "cell-friendly" deoxygenating and reducing conditions.

Original languageEnglish (US)
JournalCurrent Protocols in Protein Science
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Fluorescence microscopy
Fluorescent Dyes
Fluorescence Microscopy
Fluorophores
Cells
Proteins
Fluorescence
Bacterial Toxins
Imaging techniques
Cell membranes
Labeling
Microscopy
Microscopic examination
Molecular Weight
Molecular weight
Cell Membrane
Observation
Cell Line
streptolysin O

Keywords

  • Cell delivery
  • Fluorescence microscopy
  • Microscopy
  • Single molecule
  • Streptolysin O
  • Super-resolution

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

Cite this

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