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

Keywords

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

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

Fingerprint Dive into the research topics of 'Delivery of Fluorescent Probes Using Streptolysin O for Fluorescence Microscopy of Living Cells'. Together they form a unique fingerprint.

  • Cite this